Pascale Reverdiau - Academia.edu (original) (raw)

Papers by Pascale Reverdiau

Research paper thumbnail of Effects of Phosphatidic Acid on Cytokinin Signal Transduction in Periwinkle Cells

Journal of Plant Growth Regulation, Aug 7, 2008

In periwinkle cell suspensions, the accurate quantification of gene expression through real-time ... more In periwinkle cell suspensions, the accurate quantification of gene expression through real-time RT-PCR showed that two type-A response regulators (RR-A), considered primary cytokinin (CK)-responsive genes, were differentially regulated after CK treatment. Specific inhibition of phospholipase D (PLD)-dependent phosphatidic acid (PA) production by primary alcohols reduced significantly the transcript level of one gene in response to CK, although the other gene was unaffected. Moreover, this inhibitory effect on gene transcript level could be antagonized by exogenous supply of PA. These results suggest that PA, likely released from the membrane by PLD activity, could operate in the early steps of CK signalling in periwinkle cells.

Research paper thumbnail of Influence of MMP-2 and MMP-9 promoter polymorphisms on gene expression and clinical outcome of non-small cell lung cancer

Lung Cancer, May 1, 2007

Matrix metalloproteinases (MMP) including MMP-2 and MMP-9 play a major role in tumour invasion by... more Matrix metalloproteinases (MMP) including MMP-2 and MMP-9 play a major role in tumour invasion by proteolysing the extracellular matrix. Their activation, particularly that of MMP-9, is partly dependent on plasmin that is inhibited by TFPI-2 (tissue factor pathway inhibitor-2), a serine protease inhibitor whose gene expression is decreased in about one-third of non-small cell lung cancers (NSCLC). In addition, MMP-2 and MMP-9 are essential in the development of NSCLC and can be regulated by functional promoter polymorphisms. In this study, the -1306C/T MMP-2, -735C/T MMP-2 and -1562C/T MMP-9 polymorphisms were analysed in 90 NSCLC patients and 90 controls. In addition, the promoter region of the TFPI-2 gene was screened for sequence variations in both groups by DHPLC. A -167G/A polymorphism was identified in 3% of controls whereas none of the 90 patients exhibited this genetic variation in the TFPI-2 promoter region. Moreover, no difference in -1306C/T MMP-2, -735C/T MMP-2 and -1562C/T MMP-9 genotypes was found between cases and controls. However, the homozygous -1562CC MMP-9 genotype was more frequent in patients with squamous cell carcinoma than in controls (p=0.018). When genotype distributions were compared to MMP-2 and MMP-9 gene expression in tumours, no relationship was found with the -1306 MMP-2 and -1562 MMP-9 polymorphisms. In contrast, tumour MMP-2 gene expression was lower in homozygous -735CC patients than in those with CT or TT genotypes. In addition, the survival time was longer in patients with the MMP-2 -735T allele than in those with the CC genotype (p=0.02). The relative risk of death was increased 2.6-fold in -735CC patients (p=0.045; 95% CI=1.0-6.7). The results of this study suggest that the -735C/T MMP-2 polymorphism might be an independent prognostic marker in NSCLC, but this should be confirmed in a larger cohort of patients.

Research paper thumbnail of Peptides dérivés d'une beta-défensine 11 aviaire et leurs utilisations

Research paper thumbnail of Tfpi-2, An Inhibitor Of Proteases And Tumoral Invasion | Le Tfpi-2, Un Inhibiteur De Protéases Et De L'Invasion Tumorale

Le TFPI-2 (tissue factor pathway inhibitor 2), caractérisé dans les années 1990, est identique à ... more Le TFPI-2 (tissue factor pathway inhibitor 2), caractérisé dans les années 1990, est identique à la protéine placentaire 5 ou PP5 [1], initialement isolée à partir du placenta humain [2]. Cette glycoprotéine matricielle, appelée aussi matrix-associated serine protease inhibitor (MSPI) par Rao et al. [3, 4], est un inhibiteur de protéases à site sérine de type Kunitz dont l'implication dans différents processus physiologiques et pathologiques se révèle particulièrement importante. En effet, le TFPI-2, synthétisé par de nombreuses cellules normales et certaines cellules tumorales, serait impliqué dans la régulation de la coagulation sanguine et surtout de la migration cellulaire, de l'invasion tumorale, de l'apoptose mais également de l'athérosclérose.

Research paper thumbnail of Structure, function, and evolution of Gga -AvBD11, the archetype of the structural avian-double-β-defensin family

Proceedings of the National Academy of Sciences, 2019

Out of the 14 avian β-defensins identified in the Gallus gallus genome, only 3 are present in the... more Out of the 14 avian β-defensins identified in the Gallus gallus genome, only 3 are present in the chicken egg, including the egg-specific avian β-defensin 11 ( Gga -AvBD11). Given its specific localization and its established antibacterial activity, Gga -AvBD11 appears to play a protective role in embryonic development. Gga -AvBD11 is an atypical double-sized defensin, predicted to possess 2 motifs related to β-defensins and 6 disulfide bridges. The 3-dimensional NMR structure of the purified Gga- AvBD11 is a compact fold composed of 2 packed β-defensin domains. This fold is the archetype of a structural family, dubbed herein as avian-double-β-defensins (Av-DBD). We speculate that AvBD11 emanated from a monodomain gene ancestor and that similar events might have occurred in arthropods, leading to another structural family of less compact DBDs. We show that Gga -AvBD11 displays antimicrobial activities against gram-positive and gram-negative bacterial pathogens, the avian protozoan E...

Research paper thumbnail of Expression and methylation status of tissue factor pathway inhibitor-2 gene in non-small-cell lung cancer

British Journal of Cancer, 2005

Tissue factor pathway inhibitor-2 (TFPI-2) is a Kunitz-type serine proteinase inhibitor that inhi... more Tissue factor pathway inhibitor-2 (TFPI-2) is a Kunitz-type serine proteinase inhibitor that inhibits plasmin-dependent activation of several metalloproteinases. Downregulation of TFPI-2 could thus enhance the invasive potential of neoplastic cells in several cancers, including lung cancer. In this study, TFPI-2 mRNA was measured using a real-time PCR method in tumours of 59 patients with nonsmall-cell lung cancer (NSCLC). Tumour TFPI-2 mRNA levels appeared well correlated with protein expression evaluated by immunohistochemistry and were 4-120 times lower compared to those of nonaffected lung tissue in 22 cases (37%). Hypermethylation of the TFPI-2 gene promoter was demonstrated by restriction enzyme-polymerase chain reaction in 12 of 40 cases of NSCLC (30%), including nine of 17 for whom tumour TFPI-2 gene expression was lower than in noncancerous tissue. In contrast, this epigenetic modification was shown in only three of 23 tumours in which no decrease in TFPI-2 synthesis was found (P ¼ 0.016). Decreased TFPI-2 gene expression and hypermethylation were more frequently associated with stages III or IV NSCLC (eight out of 10, P ¼ 0.02) and the TFPI-2 gene promoter was more frequently hypermethylated in patients with lymph node metastases (eight out of 16, P ¼ 0.02). These results suggest that silencing of the TFPI-2 gene by hypermethylation might contribute to tumour progression in NSCLC.

Research paper thumbnail of P-008 Tumour expression and activity of MMP-2 and MMP-9metalloproteinases in non-small cell lung cancer according to TFPI-2 gene expression

Lung Cancer, 2005

Posters / Basic science~Ceil and motecular biology vrtto cell preliferation, this response was mo... more Posters / Basic science~Ceil and motecular biology vrtto cell preliferation, this response was most apparent for NCI+H.60 with low endogenous USF-2 Co <0 005) reducing the mean call doubling time From 19 to 16 hours Dominant-negative USF-2 mutants had no significant effect end USF-2 everexpression did not effect proliferation of SCLC (high endogenous USF-2) or MCF-7 breast adenocarcinoma cells (LISF-2 inactive) Conduslona: These data suggest that USF-2 not only represents a relatively eady molecular marker for the development of bronchial dysp~asia and non-adenocarcinema lung cancer, but may also play a role in bronchial ca~nogenos~s. Although USF 2 is downrogulated or inactivated in breast cancer, we suggest that this transcription factor plays a different role in lung cancer, where It can promote cell proliferation. However. the genes through which this tTansc~ptional control of cell growth by USF 2 is mediated remain to be determined. [~ Role of vascular endothelial grow~l factor (VEGF) In the diagnosis of malignant plaural effusions

Research paper thumbnail of 054 Le Tissue Factor Pathway lnhibitor-2, un nouveau gène suppresseur de tumeur ?

Revue des Maladies Respiratoires, 2005

053 L'inhibition de la phosphorylation des chaînes légères régulatrices de la myosine induit I'ap... more 053 L'inhibition de la phosphorylation des chaînes légères régulatrices de la myosine induit I'apoptose et 1'autophagie.des cellules épithéliales d'adénocarcinome pulmonaire humain ' INSERM U700,

Research paper thumbnail of Tumor-stromal cell interactions modulate metalloproteinase and kalli Krein expression in direct and indirect co-culture cell models

Revue des Maladies Respiratoires, 2008

The crosstalk between tumor cells and surrounding stromal fibroblasts is now considered crucial f... more The crosstalk between tumor cells and surrounding stromal fibroblasts is now considered crucial for cancer progression, particularly in invasive tumors such as lung carcinomas. Tumorstromal cell interactions might provide signal for regulating protease and protease inhibitor secretion in the tumoral microenvironment and modulate extracellular matrix (ECM) proteolysis and then tumor invasion. The aim of this study was to develop co-culture models with cancer cells, derived from a non-small cell lung carcinoma (NSCLC), and fibroblast cells. Expression of several MMPs, kallicrein 6 (KLK) and Tissue Factor Pathway inhibitor 2 (TFPI-2) was then measured in these models. Material and methods: Two in vitro co-culture models were developed to evaluate the effects of direct or indirect contact between NSCLC NCI-H460 cells and CCD19-Lu fibroblast cells. In direct co-culture, both cells (ratio 1 :1) were cultured for 24 h in serum free medium. In indirect co-culture, conditioned media were collected from either confluent tumoral cells or fibroblasts grown in serum free medium during 24 h. Transcript levels of MMP-1,-2,-3,-9,-13 and-14, EMMPRIN (Extracellular Matrix MetalloPRoteinase Inducer), KLK6 and TFPI-2 were measured using specific quantitative real-time RT-PCR. Protein expressions were evaluated by Western Blotting and immunofluorescence staining. Results: We found a 3-fold and 8-fold increase of MMP-3 and MMP-9 expression respectively in the direct co-culture compared to cells grown alone. Although the level was lower, KLK6 mRNA was also enhanced in direct co-culture. In indirect co-culture with CCD19Lu cultured with NCI-H460 conditioned medium, we observed an increase in MMP-1,-3,-9 and TFPI-2 transcripts. Except for MMP3 and KLK6, no difference in transcripts level were observed in the other indirect co-culture model, i. e NCI-H460 grown in CCD19Lu conditioned medium. Conclusion: Our results indicate that direct or indirect contacts between tumors and surrounding fibroblasts modulate the expression of various proteinases. This effect might be mediated by soluble or/ and cell surface factors. Further investigations will be required to identify them.

Research paper thumbnail of Monitoring of tumor progression using bioluminescence imaging in a nude mice orthotopic model of human small cell lung cancer

Revue des Maladies Respiratoires, 2008

Research paper thumbnail of Beneficial role of overexpression of TFPI-2 on tumour progression in human small cell lung cancer

FEBS Open Bio, 2013

Tissue factor pathway inhibitor-2 (TFPI-2) is a potent inhibitor of plasmin, a protease which is ... more Tissue factor pathway inhibitor-2 (TFPI-2) is a potent inhibitor of plasmin, a protease which is involved in tumour progression by activating (MMPs). This therefore makes TFPI-2 a potential inhibitor of invasiveness and the development of metastases. In this study, low levels of TFPI-2 expression were found in 65% of patients with small cell lung cancer (SCLC), the most aggressive type of lung cancer. To study the impact of TFPI-2 in tumour progression, TFPI-2 was overexpressed in NCI-H209 SCLC cells which were orthotopically implanted in nude mice. Investigations showed that TFPI-2 inhibited lung tumour growth. Such inhibition could be explained in vitro by a decrease in tumour cell viability, blockade of G1 / S phase cell cycle transition and an increase in apoptosis shown in NCI-H209 cells expressing TFPI-2. We also demonstrated that TFPI-2 upregulation in NCI-H209 cells decreased MMP expression, particularly by downregulating MMP-1 and MMP-3. Moreover, TFPI-2 inhibited phosphorylation of the MAPK signalling pathway proteins involved in the induction of MMP transcripts, among which MMP-1 was predominant in SCLC tissues and was inversely expressed with TFPI-2 in 35% of cases. These results suggest that downregulation of TFPI-2 expression could favour the development of SCLC.

Research paper thumbnail of Fast Detection of Tissue Factor and Tissue Factor Pathway Inhibitor Messenger RNA in Endothelial Cells and Monocytes by Sensitive Reverse Transcription–Polymerase Chain Reaction

Thrombosis Research, 1999

out stimulation by endotoxin was mainly related to cell adhesion. TFPI-1 mRNA was constitutively ... more out stimulation by endotoxin was mainly related to cell adhesion. TFPI-1 mRNA was constitutively We developed fast and sensitive reverse transcripexpressed in endothelial cells and was detected in tion-polymerase chain reaction (RT-PCR) proceonly 5.10 2 unstimulated cells and 10 2 phorbol esterdures to study the expression of tissue factor (TF) stimulated cells. Expression was increased upon and tissue factor pathway inhibitor (TFPI-1) mRNA stimulation with phorbol ester. With this techin human endothelial cells and monocytes. The nique, TFPI-1 mRNA in monocytes was rather low sensitivity of the technique was checked by pereven when cells were stimulated with phorbol ester forming RT-PCR with limited numbers of cells. Cells were stimulated either with tumor necrosis or after adhesion.

Research paper thumbnail of The Airways, a Novel Route for Delivering Monoclonal Antibodies to Treat Lung Tumors

Pharmaceutical Research, 2011

Lung cancer is the leading cause of cancer-related death worldwide. The efficacy of current syste... more Lung cancer is the leading cause of cancer-related death worldwide. The efficacy of current systemic treatments is limited, with major side effects and only modest survival improvements. Aerosols routinely used to deliver drugs into the lung for treating infectious and inflammatory lung diseases have never been used to deliver monoclonal antibodies to treat lung cancer. We have shown that cetuximab, a chimeric anticancer anti-EGFR mAb, is suitable for airway delivery as it resists the physical constraints of aerosolization, and have evaluated the aerosol delivery of cetuximab in vivo. We developed an animal model of lung tumor sensitive to cetuximab by injecting Balb/c Nude mice intratracheally with A431 cells plus 10 mM EDTA and analyzed the distribution, pharmacokinetics and antitumor efficacy of cetuximab aerosolized into the respiratory tract. Aerosolized IgG accumulated durably in the lungs and the tumor, but passed poorly and slowly into the systemic circulation. Aerosolized cetuximab also limited the growth of the mouse tumor. Thus, administering anticancer mAbs via the airways is effective and may limit systemic side effects. Delivery of aerosolized-mAbs via the airways deserves further evaluation for treating lung cancers.

Research paper thumbnail of Inhibition of cervical cancer cell growth by human papillomavirus virus–like particles packaged with human papillomavirus oncoprotein short hairpin RNAs

Molecular Cancer Therapeutics, 2009

Overexpression of human papillomavirus (HPV E6 and HPV E7) oncogenes in human cervical cells resu... more Overexpression of human papillomavirus (HPV E6 and HPV E7) oncogenes in human cervical cells results in the development of cancer, and E6 and E7 proteins are therefore targets for preventing cervical cancer progression. Here, we describe the silencing of E6 and E7 expression in cervical carcinoma cells by RNA interference. In order to increase the efficacy of the RNA interference, HPV pseudovirions coding for a short hairpin RNA (shRNA) sequence were produced. The results indicated the degradation of E6 and E7 mRNAs when shRNA against E6 or E7 were delivered by pseudovirions in HPV-positive cells (CaSki and TC1 cells). E6 silencing resulted in the accumulation of cellular p53 and reduced cell viability. More significant cell death was observed when E7 expression was suppressed. Silencing E6 and E7 and the consequences for cancer cell growth were also investigated in vivo in mice using the capacity of murine TC1 cells expressing HPV-16 E6 and E7 oncogenes to induce fast-growing tumor...

Research paper thumbnail of Improved PCR Method for Amplification of GC-Rich DNA Sequences

Molecular Biotechnology, 2005

Research paper thumbnail of Monitoring of tumour progression using bioluminescence imaging and computed tomography scanning in a nude mouse orthotopic model of human small cell lung cancer

Lung Cancer, 2012

Human small cell lung carcinoma (SCLC) is the most aggressive type of lung cancer but no clinical... more Human small cell lung carcinoma (SCLC) is the most aggressive type of lung cancer but no clinically relevant animal model has been developed to date. Such a model would be valuable to study the molecular aspects of tumour progression and to test the effectiveness of new treatment agents. We generated a reproducible and reliable nude mouse orthotopic model of human SCLC with NCI-H209 tumour cells genetically modified to express firefly luciferase. Cells were analysed for long-term stability of bioluminescence and a clone was passaged twice subcutaneously to enhance tumorigenicity. Cells resuspended in Matrigel and/or EDTA RPMI medium containing a (99m)Tc-labelled tin colloid used as tracer were implanted intrabronchially with a catheter inserted into the trachea and positioned in the main bronchus using X-ray-guided imaging. Deposition of cells into the lung was then assessed by scintigraphy. The growth of the primary tumour was sensitively and non-invasively followed by bioluminescence imaging that allowed real-time monitoring of tumour progression in the same animals over a 2-12-week period. Additional 3D bioluminescence imaging and computed tomography scanning were used to document tumour location and measurements that were confirmed by histological analyses. In conclusion, this original nude mouse orthotopic model resembles various stages of human small cell lung cancer, and therefore could be used to evaluate new treatment strategies.

Research paper thumbnail of Influence of MMP-2 and MMP-9 promoter polymorphisms on gene expression and clinical outcome of non-small cell lung cancer

Lung Cancer, 2007

Matrix metalloproteinases (MMP) including MMP-2 and MMP-9 play a major role in tumour invasion by... more Matrix metalloproteinases (MMP) including MMP-2 and MMP-9 play a major role in tumour invasion by proteolysing the extracellular matrix. Their activation, particularly that of MMP-9, is partly dependent on plasmin that is inhibited by TFPI-2 (tissue factor pathway inhibitor-2), a serine protease inhibitor whose gene expression is decreased in about one-third of non-small cell lung cancers (NSCLC). In addition, MMP-2 and MMP-9 are essential in the development of NSCLC and can be regulated by functional promoter polymorphisms. In this study, the -1306C/T MMP-2, -735C/T MMP-2 and -1562C/T MMP-9 polymorphisms were analysed in 90 NSCLC patients and 90 controls. In addition, the promoter region of the TFPI-2 gene was screened for sequence variations in both groups by DHPLC. A -167G/A polymorphism was identified in 3% of controls whereas none of the 90 patients exhibited this genetic variation in the TFPI-2 promoter region. Moreover, no difference in -1306C/T MMP-2, -735C/T MMP-2 and -1562C/T MMP-9 genotypes was found between cases and controls. However, the homozygous -1562CC MMP-9 genotype was more frequent in patients with squamous cell carcinoma than in controls (p=0.018). When genotype distributions were compared to MMP-2 and MMP-9 gene expression in tumours, no relationship was found with the -1306 MMP-2 and -1562 MMP-9 polymorphisms. In contrast, tumour MMP-2 gene expression was lower in homozygous -735CC patients than in those with CT or TT genotypes. In addition, the survival time was longer in patients with the MMP-2 -735T allele than in those with the CC genotype (p=0.02). The relative risk of death was increased 2.6-fold in -735CC patients (p=0.045; 95% CI=1.0-6.7). The results of this study suggest that the -735C/T MMP-2 polymorphism might be an independent prognostic marker in NSCLC, but this should be confirmed in a larger cohort of patients.

Research paper thumbnail of Effects of Phosphatidic Acid on Cytokinin Signal Transduction in Periwinkle Cells

Journal of Plant Growth Regulation, 2008

In periwinkle cell suspensions, the accurate quantification of gene expression through real-time ... more In periwinkle cell suspensions, the accurate quantification of gene expression through real-time RT-PCR showed that two type-A response regulators (RR-A), considered primary cytokinin (CK)-responsive genes, were differentially regulated after CK treatment. Specific inhibition of phospholipase D (PLD)-dependent phosphatidic acid (PA) production by primary alcohols reduced significantly the transcript level of one gene in response to CK, although the other gene was unaffected. Moreover, this inhibitory effect on gene transcript level could be antagonized by exogenous supply of PA. These results suggest that PA, likely released from the membrane by PLD activity, could operate in the early steps of CK signalling in periwinkle cells.

Research paper thumbnail of CCL5-enhanced human immature dendritic cell migration through the basement membrane in vitro depends on matrix metalloproteinase-9

Journal of Leukocyte Biology, 2006

The proinflammatory chemokine CC chemokine ligand 5 (CCL5) is a potent chemoattractant of immatur... more The proinflammatory chemokine CC chemokine ligand 5 (CCL5) is a potent chemoattractant of immature dendritic cells (iDCs). It remains to be elucidated whether CCL5 may also enhance iDC migration through the basement membrane by affecting matrix metalloproteinase (MMP)-9 secretion. In this study, iDCs were differentiated in vitro from human monocytes of healthy donors. Zymographic analysis of cellular membranes of nontreated iDCs revealed a basal secretion of the pro- and active MMP-9, whereas only pro-MMP-9 was detected in conditioned media. Increasing concentrations of CCL5 significantly enhanced MMP-9 secretion by iDCs, peaking at 100 ng/ml, which optimally increased iDC migration through a reconstituted basement membrane (Matrigel™) in vitro. The CCL5-enhanced secretion of MMP-9 occurred early (2 h) and was maintained at least for 10 h. A significant increase in MMP-9 mRNA synthesis was detected by reverse transcriptase-polymerase chain reaction, only at 6 h of CCL5 treatment, wh...

Research paper thumbnail of TFPI-2 silencing increases tumour progression and promotes metalloproteinase 1 and 3 induction through tumour-stromal cell interactions

Journal of Cellular and Molecular Medicine, 2011

Tumour progression is a complex multistep process that depends on an evolving crosstalk between c... more Tumour progression is a complex multistep process that depends on an evolving crosstalk between cancer cells and the surrounding stromal tissue. The microenvironment is now recognized as having a pivotal role in promoting cancer initiation, progression and dissemination to form metastases [1]. The invasion process involves extracellular matrix (ECM)-degrading proteases, particularly matrix metalloproteinases (MMPs), that have been shown to be highly expressed and activated in the tumour microenvironment [2], especially in highly aggressive malignant tumours [3, 4]. Activated fibroblasts, the major cell component of the microenvironment, actively contribute to tumour invasiveness by secreting a consistent amount of MMPs at the tumour-stroma interface. Moreover, this MMP synthesis could be increased by the ECM metalloproteinase inducer (EMMPRIN) expressed by cancer cells. Activation of zymogens (pro-MMPs) in the extracellular environment needs proteolytic cleavage of the aminoterminal prodomain [5] that depends on serine proteases, such as trypsin and plasmin, and also involves activated MMPs or membrane-anchored matrix metalloproteinases (MT-MMPs). Apart from tissue inhibitors of metalloproteinases (TIMPs) that are specific regulators of MMP activity, TFPI-2 (tissue factor pathway inhibitor-2), an inhibitor of serine proteases-particularly plasmin-could also regulate the activation of MMPs [6, 7], thus regulating ECM degradation and tumour cell invasion. TFPI-2 silencing increases tumour progression and promotes metalloproteinase 1 and 3 induction through tumour-stromal cell interactions

Research paper thumbnail of Effects of Phosphatidic Acid on Cytokinin Signal Transduction in Periwinkle Cells

Journal of Plant Growth Regulation, Aug 7, 2008

In periwinkle cell suspensions, the accurate quantification of gene expression through real-time ... more In periwinkle cell suspensions, the accurate quantification of gene expression through real-time RT-PCR showed that two type-A response regulators (RR-A), considered primary cytokinin (CK)-responsive genes, were differentially regulated after CK treatment. Specific inhibition of phospholipase D (PLD)-dependent phosphatidic acid (PA) production by primary alcohols reduced significantly the transcript level of one gene in response to CK, although the other gene was unaffected. Moreover, this inhibitory effect on gene transcript level could be antagonized by exogenous supply of PA. These results suggest that PA, likely released from the membrane by PLD activity, could operate in the early steps of CK signalling in periwinkle cells.

Research paper thumbnail of Influence of MMP-2 and MMP-9 promoter polymorphisms on gene expression and clinical outcome of non-small cell lung cancer

Lung Cancer, May 1, 2007

Matrix metalloproteinases (MMP) including MMP-2 and MMP-9 play a major role in tumour invasion by... more Matrix metalloproteinases (MMP) including MMP-2 and MMP-9 play a major role in tumour invasion by proteolysing the extracellular matrix. Their activation, particularly that of MMP-9, is partly dependent on plasmin that is inhibited by TFPI-2 (tissue factor pathway inhibitor-2), a serine protease inhibitor whose gene expression is decreased in about one-third of non-small cell lung cancers (NSCLC). In addition, MMP-2 and MMP-9 are essential in the development of NSCLC and can be regulated by functional promoter polymorphisms. In this study, the -1306C/T MMP-2, -735C/T MMP-2 and -1562C/T MMP-9 polymorphisms were analysed in 90 NSCLC patients and 90 controls. In addition, the promoter region of the TFPI-2 gene was screened for sequence variations in both groups by DHPLC. A -167G/A polymorphism was identified in 3% of controls whereas none of the 90 patients exhibited this genetic variation in the TFPI-2 promoter region. Moreover, no difference in -1306C/T MMP-2, -735C/T MMP-2 and -1562C/T MMP-9 genotypes was found between cases and controls. However, the homozygous -1562CC MMP-9 genotype was more frequent in patients with squamous cell carcinoma than in controls (p=0.018). When genotype distributions were compared to MMP-2 and MMP-9 gene expression in tumours, no relationship was found with the -1306 MMP-2 and -1562 MMP-9 polymorphisms. In contrast, tumour MMP-2 gene expression was lower in homozygous -735CC patients than in those with CT or TT genotypes. In addition, the survival time was longer in patients with the MMP-2 -735T allele than in those with the CC genotype (p=0.02). The relative risk of death was increased 2.6-fold in -735CC patients (p=0.045; 95% CI=1.0-6.7). The results of this study suggest that the -735C/T MMP-2 polymorphism might be an independent prognostic marker in NSCLC, but this should be confirmed in a larger cohort of patients.

Research paper thumbnail of Peptides dérivés d'une beta-défensine 11 aviaire et leurs utilisations

Research paper thumbnail of Tfpi-2, An Inhibitor Of Proteases And Tumoral Invasion | Le Tfpi-2, Un Inhibiteur De Protéases Et De L'Invasion Tumorale

Le TFPI-2 (tissue factor pathway inhibitor 2), caractérisé dans les années 1990, est identique à ... more Le TFPI-2 (tissue factor pathway inhibitor 2), caractérisé dans les années 1990, est identique à la protéine placentaire 5 ou PP5 [1], initialement isolée à partir du placenta humain [2]. Cette glycoprotéine matricielle, appelée aussi matrix-associated serine protease inhibitor (MSPI) par Rao et al. [3, 4], est un inhibiteur de protéases à site sérine de type Kunitz dont l'implication dans différents processus physiologiques et pathologiques se révèle particulièrement importante. En effet, le TFPI-2, synthétisé par de nombreuses cellules normales et certaines cellules tumorales, serait impliqué dans la régulation de la coagulation sanguine et surtout de la migration cellulaire, de l'invasion tumorale, de l'apoptose mais également de l'athérosclérose.

Research paper thumbnail of Structure, function, and evolution of Gga -AvBD11, the archetype of the structural avian-double-β-defensin family

Proceedings of the National Academy of Sciences, 2019

Out of the 14 avian β-defensins identified in the Gallus gallus genome, only 3 are present in the... more Out of the 14 avian β-defensins identified in the Gallus gallus genome, only 3 are present in the chicken egg, including the egg-specific avian β-defensin 11 ( Gga -AvBD11). Given its specific localization and its established antibacterial activity, Gga -AvBD11 appears to play a protective role in embryonic development. Gga -AvBD11 is an atypical double-sized defensin, predicted to possess 2 motifs related to β-defensins and 6 disulfide bridges. The 3-dimensional NMR structure of the purified Gga- AvBD11 is a compact fold composed of 2 packed β-defensin domains. This fold is the archetype of a structural family, dubbed herein as avian-double-β-defensins (Av-DBD). We speculate that AvBD11 emanated from a monodomain gene ancestor and that similar events might have occurred in arthropods, leading to another structural family of less compact DBDs. We show that Gga -AvBD11 displays antimicrobial activities against gram-positive and gram-negative bacterial pathogens, the avian protozoan E...

Research paper thumbnail of Expression and methylation status of tissue factor pathway inhibitor-2 gene in non-small-cell lung cancer

British Journal of Cancer, 2005

Tissue factor pathway inhibitor-2 (TFPI-2) is a Kunitz-type serine proteinase inhibitor that inhi... more Tissue factor pathway inhibitor-2 (TFPI-2) is a Kunitz-type serine proteinase inhibitor that inhibits plasmin-dependent activation of several metalloproteinases. Downregulation of TFPI-2 could thus enhance the invasive potential of neoplastic cells in several cancers, including lung cancer. In this study, TFPI-2 mRNA was measured using a real-time PCR method in tumours of 59 patients with nonsmall-cell lung cancer (NSCLC). Tumour TFPI-2 mRNA levels appeared well correlated with protein expression evaluated by immunohistochemistry and were 4-120 times lower compared to those of nonaffected lung tissue in 22 cases (37%). Hypermethylation of the TFPI-2 gene promoter was demonstrated by restriction enzyme-polymerase chain reaction in 12 of 40 cases of NSCLC (30%), including nine of 17 for whom tumour TFPI-2 gene expression was lower than in noncancerous tissue. In contrast, this epigenetic modification was shown in only three of 23 tumours in which no decrease in TFPI-2 synthesis was found (P ¼ 0.016). Decreased TFPI-2 gene expression and hypermethylation were more frequently associated with stages III or IV NSCLC (eight out of 10, P ¼ 0.02) and the TFPI-2 gene promoter was more frequently hypermethylated in patients with lymph node metastases (eight out of 16, P ¼ 0.02). These results suggest that silencing of the TFPI-2 gene by hypermethylation might contribute to tumour progression in NSCLC.

Research paper thumbnail of P-008 Tumour expression and activity of MMP-2 and MMP-9metalloproteinases in non-small cell lung cancer according to TFPI-2 gene expression

Lung Cancer, 2005

Posters / Basic science~Ceil and motecular biology vrtto cell preliferation, this response was mo... more Posters / Basic science~Ceil and motecular biology vrtto cell preliferation, this response was most apparent for NCI+H.60 with low endogenous USF-2 Co <0 005) reducing the mean call doubling time From 19 to 16 hours Dominant-negative USF-2 mutants had no significant effect end USF-2 everexpression did not effect proliferation of SCLC (high endogenous USF-2) or MCF-7 breast adenocarcinoma cells (LISF-2 inactive) Conduslona: These data suggest that USF-2 not only represents a relatively eady molecular marker for the development of bronchial dysp~asia and non-adenocarcinema lung cancer, but may also play a role in bronchial ca~nogenos~s. Although USF 2 is downrogulated or inactivated in breast cancer, we suggest that this transcription factor plays a different role in lung cancer, where It can promote cell proliferation. However. the genes through which this tTansc~ptional control of cell growth by USF 2 is mediated remain to be determined. [~ Role of vascular endothelial grow~l factor (VEGF) In the diagnosis of malignant plaural effusions

Research paper thumbnail of 054 Le Tissue Factor Pathway lnhibitor-2, un nouveau gène suppresseur de tumeur ?

Revue des Maladies Respiratoires, 2005

053 L'inhibition de la phosphorylation des chaînes légères régulatrices de la myosine induit I'ap... more 053 L'inhibition de la phosphorylation des chaînes légères régulatrices de la myosine induit I'apoptose et 1'autophagie.des cellules épithéliales d'adénocarcinome pulmonaire humain ' INSERM U700,

Research paper thumbnail of Tumor-stromal cell interactions modulate metalloproteinase and kalli Krein expression in direct and indirect co-culture cell models

Revue des Maladies Respiratoires, 2008

The crosstalk between tumor cells and surrounding stromal fibroblasts is now considered crucial f... more The crosstalk between tumor cells and surrounding stromal fibroblasts is now considered crucial for cancer progression, particularly in invasive tumors such as lung carcinomas. Tumorstromal cell interactions might provide signal for regulating protease and protease inhibitor secretion in the tumoral microenvironment and modulate extracellular matrix (ECM) proteolysis and then tumor invasion. The aim of this study was to develop co-culture models with cancer cells, derived from a non-small cell lung carcinoma (NSCLC), and fibroblast cells. Expression of several MMPs, kallicrein 6 (KLK) and Tissue Factor Pathway inhibitor 2 (TFPI-2) was then measured in these models. Material and methods: Two in vitro co-culture models were developed to evaluate the effects of direct or indirect contact between NSCLC NCI-H460 cells and CCD19-Lu fibroblast cells. In direct co-culture, both cells (ratio 1 :1) were cultured for 24 h in serum free medium. In indirect co-culture, conditioned media were collected from either confluent tumoral cells or fibroblasts grown in serum free medium during 24 h. Transcript levels of MMP-1,-2,-3,-9,-13 and-14, EMMPRIN (Extracellular Matrix MetalloPRoteinase Inducer), KLK6 and TFPI-2 were measured using specific quantitative real-time RT-PCR. Protein expressions were evaluated by Western Blotting and immunofluorescence staining. Results: We found a 3-fold and 8-fold increase of MMP-3 and MMP-9 expression respectively in the direct co-culture compared to cells grown alone. Although the level was lower, KLK6 mRNA was also enhanced in direct co-culture. In indirect co-culture with CCD19Lu cultured with NCI-H460 conditioned medium, we observed an increase in MMP-1,-3,-9 and TFPI-2 transcripts. Except for MMP3 and KLK6, no difference in transcripts level were observed in the other indirect co-culture model, i. e NCI-H460 grown in CCD19Lu conditioned medium. Conclusion: Our results indicate that direct or indirect contacts between tumors and surrounding fibroblasts modulate the expression of various proteinases. This effect might be mediated by soluble or/ and cell surface factors. Further investigations will be required to identify them.

Research paper thumbnail of Monitoring of tumor progression using bioluminescence imaging in a nude mice orthotopic model of human small cell lung cancer

Revue des Maladies Respiratoires, 2008

Research paper thumbnail of Beneficial role of overexpression of TFPI-2 on tumour progression in human small cell lung cancer

FEBS Open Bio, 2013

Tissue factor pathway inhibitor-2 (TFPI-2) is a potent inhibitor of plasmin, a protease which is ... more Tissue factor pathway inhibitor-2 (TFPI-2) is a potent inhibitor of plasmin, a protease which is involved in tumour progression by activating (MMPs). This therefore makes TFPI-2 a potential inhibitor of invasiveness and the development of metastases. In this study, low levels of TFPI-2 expression were found in 65% of patients with small cell lung cancer (SCLC), the most aggressive type of lung cancer. To study the impact of TFPI-2 in tumour progression, TFPI-2 was overexpressed in NCI-H209 SCLC cells which were orthotopically implanted in nude mice. Investigations showed that TFPI-2 inhibited lung tumour growth. Such inhibition could be explained in vitro by a decrease in tumour cell viability, blockade of G1 / S phase cell cycle transition and an increase in apoptosis shown in NCI-H209 cells expressing TFPI-2. We also demonstrated that TFPI-2 upregulation in NCI-H209 cells decreased MMP expression, particularly by downregulating MMP-1 and MMP-3. Moreover, TFPI-2 inhibited phosphorylation of the MAPK signalling pathway proteins involved in the induction of MMP transcripts, among which MMP-1 was predominant in SCLC tissues and was inversely expressed with TFPI-2 in 35% of cases. These results suggest that downregulation of TFPI-2 expression could favour the development of SCLC.

Research paper thumbnail of Fast Detection of Tissue Factor and Tissue Factor Pathway Inhibitor Messenger RNA in Endothelial Cells and Monocytes by Sensitive Reverse Transcription–Polymerase Chain Reaction

Thrombosis Research, 1999

out stimulation by endotoxin was mainly related to cell adhesion. TFPI-1 mRNA was constitutively ... more out stimulation by endotoxin was mainly related to cell adhesion. TFPI-1 mRNA was constitutively We developed fast and sensitive reverse transcripexpressed in endothelial cells and was detected in tion-polymerase chain reaction (RT-PCR) proceonly 5.10 2 unstimulated cells and 10 2 phorbol esterdures to study the expression of tissue factor (TF) stimulated cells. Expression was increased upon and tissue factor pathway inhibitor (TFPI-1) mRNA stimulation with phorbol ester. With this techin human endothelial cells and monocytes. The nique, TFPI-1 mRNA in monocytes was rather low sensitivity of the technique was checked by pereven when cells were stimulated with phorbol ester forming RT-PCR with limited numbers of cells. Cells were stimulated either with tumor necrosis or after adhesion.

Research paper thumbnail of The Airways, a Novel Route for Delivering Monoclonal Antibodies to Treat Lung Tumors

Pharmaceutical Research, 2011

Lung cancer is the leading cause of cancer-related death worldwide. The efficacy of current syste... more Lung cancer is the leading cause of cancer-related death worldwide. The efficacy of current systemic treatments is limited, with major side effects and only modest survival improvements. Aerosols routinely used to deliver drugs into the lung for treating infectious and inflammatory lung diseases have never been used to deliver monoclonal antibodies to treat lung cancer. We have shown that cetuximab, a chimeric anticancer anti-EGFR mAb, is suitable for airway delivery as it resists the physical constraints of aerosolization, and have evaluated the aerosol delivery of cetuximab in vivo. We developed an animal model of lung tumor sensitive to cetuximab by injecting Balb/c Nude mice intratracheally with A431 cells plus 10 mM EDTA and analyzed the distribution, pharmacokinetics and antitumor efficacy of cetuximab aerosolized into the respiratory tract. Aerosolized IgG accumulated durably in the lungs and the tumor, but passed poorly and slowly into the systemic circulation. Aerosolized cetuximab also limited the growth of the mouse tumor. Thus, administering anticancer mAbs via the airways is effective and may limit systemic side effects. Delivery of aerosolized-mAbs via the airways deserves further evaluation for treating lung cancers.

Research paper thumbnail of Inhibition of cervical cancer cell growth by human papillomavirus virus–like particles packaged with human papillomavirus oncoprotein short hairpin RNAs

Molecular Cancer Therapeutics, 2009

Overexpression of human papillomavirus (HPV E6 and HPV E7) oncogenes in human cervical cells resu... more Overexpression of human papillomavirus (HPV E6 and HPV E7) oncogenes in human cervical cells results in the development of cancer, and E6 and E7 proteins are therefore targets for preventing cervical cancer progression. Here, we describe the silencing of E6 and E7 expression in cervical carcinoma cells by RNA interference. In order to increase the efficacy of the RNA interference, HPV pseudovirions coding for a short hairpin RNA (shRNA) sequence were produced. The results indicated the degradation of E6 and E7 mRNAs when shRNA against E6 or E7 were delivered by pseudovirions in HPV-positive cells (CaSki and TC1 cells). E6 silencing resulted in the accumulation of cellular p53 and reduced cell viability. More significant cell death was observed when E7 expression was suppressed. Silencing E6 and E7 and the consequences for cancer cell growth were also investigated in vivo in mice using the capacity of murine TC1 cells expressing HPV-16 E6 and E7 oncogenes to induce fast-growing tumor...

Research paper thumbnail of Improved PCR Method for Amplification of GC-Rich DNA Sequences

Molecular Biotechnology, 2005

Research paper thumbnail of Monitoring of tumour progression using bioluminescence imaging and computed tomography scanning in a nude mouse orthotopic model of human small cell lung cancer

Lung Cancer, 2012

Human small cell lung carcinoma (SCLC) is the most aggressive type of lung cancer but no clinical... more Human small cell lung carcinoma (SCLC) is the most aggressive type of lung cancer but no clinically relevant animal model has been developed to date. Such a model would be valuable to study the molecular aspects of tumour progression and to test the effectiveness of new treatment agents. We generated a reproducible and reliable nude mouse orthotopic model of human SCLC with NCI-H209 tumour cells genetically modified to express firefly luciferase. Cells were analysed for long-term stability of bioluminescence and a clone was passaged twice subcutaneously to enhance tumorigenicity. Cells resuspended in Matrigel and/or EDTA RPMI medium containing a (99m)Tc-labelled tin colloid used as tracer were implanted intrabronchially with a catheter inserted into the trachea and positioned in the main bronchus using X-ray-guided imaging. Deposition of cells into the lung was then assessed by scintigraphy. The growth of the primary tumour was sensitively and non-invasively followed by bioluminescence imaging that allowed real-time monitoring of tumour progression in the same animals over a 2-12-week period. Additional 3D bioluminescence imaging and computed tomography scanning were used to document tumour location and measurements that were confirmed by histological analyses. In conclusion, this original nude mouse orthotopic model resembles various stages of human small cell lung cancer, and therefore could be used to evaluate new treatment strategies.

Research paper thumbnail of Influence of MMP-2 and MMP-9 promoter polymorphisms on gene expression and clinical outcome of non-small cell lung cancer

Lung Cancer, 2007

Matrix metalloproteinases (MMP) including MMP-2 and MMP-9 play a major role in tumour invasion by... more Matrix metalloproteinases (MMP) including MMP-2 and MMP-9 play a major role in tumour invasion by proteolysing the extracellular matrix. Their activation, particularly that of MMP-9, is partly dependent on plasmin that is inhibited by TFPI-2 (tissue factor pathway inhibitor-2), a serine protease inhibitor whose gene expression is decreased in about one-third of non-small cell lung cancers (NSCLC). In addition, MMP-2 and MMP-9 are essential in the development of NSCLC and can be regulated by functional promoter polymorphisms. In this study, the -1306C/T MMP-2, -735C/T MMP-2 and -1562C/T MMP-9 polymorphisms were analysed in 90 NSCLC patients and 90 controls. In addition, the promoter region of the TFPI-2 gene was screened for sequence variations in both groups by DHPLC. A -167G/A polymorphism was identified in 3% of controls whereas none of the 90 patients exhibited this genetic variation in the TFPI-2 promoter region. Moreover, no difference in -1306C/T MMP-2, -735C/T MMP-2 and -1562C/T MMP-9 genotypes was found between cases and controls. However, the homozygous -1562CC MMP-9 genotype was more frequent in patients with squamous cell carcinoma than in controls (p=0.018). When genotype distributions were compared to MMP-2 and MMP-9 gene expression in tumours, no relationship was found with the -1306 MMP-2 and -1562 MMP-9 polymorphisms. In contrast, tumour MMP-2 gene expression was lower in homozygous -735CC patients than in those with CT or TT genotypes. In addition, the survival time was longer in patients with the MMP-2 -735T allele than in those with the CC genotype (p=0.02). The relative risk of death was increased 2.6-fold in -735CC patients (p=0.045; 95% CI=1.0-6.7). The results of this study suggest that the -735C/T MMP-2 polymorphism might be an independent prognostic marker in NSCLC, but this should be confirmed in a larger cohort of patients.

Research paper thumbnail of Effects of Phosphatidic Acid on Cytokinin Signal Transduction in Periwinkle Cells

Journal of Plant Growth Regulation, 2008

In periwinkle cell suspensions, the accurate quantification of gene expression through real-time ... more In periwinkle cell suspensions, the accurate quantification of gene expression through real-time RT-PCR showed that two type-A response regulators (RR-A), considered primary cytokinin (CK)-responsive genes, were differentially regulated after CK treatment. Specific inhibition of phospholipase D (PLD)-dependent phosphatidic acid (PA) production by primary alcohols reduced significantly the transcript level of one gene in response to CK, although the other gene was unaffected. Moreover, this inhibitory effect on gene transcript level could be antagonized by exogenous supply of PA. These results suggest that PA, likely released from the membrane by PLD activity, could operate in the early steps of CK signalling in periwinkle cells.

Research paper thumbnail of CCL5-enhanced human immature dendritic cell migration through the basement membrane in vitro depends on matrix metalloproteinase-9

Journal of Leukocyte Biology, 2006

The proinflammatory chemokine CC chemokine ligand 5 (CCL5) is a potent chemoattractant of immatur... more The proinflammatory chemokine CC chemokine ligand 5 (CCL5) is a potent chemoattractant of immature dendritic cells (iDCs). It remains to be elucidated whether CCL5 may also enhance iDC migration through the basement membrane by affecting matrix metalloproteinase (MMP)-9 secretion. In this study, iDCs were differentiated in vitro from human monocytes of healthy donors. Zymographic analysis of cellular membranes of nontreated iDCs revealed a basal secretion of the pro- and active MMP-9, whereas only pro-MMP-9 was detected in conditioned media. Increasing concentrations of CCL5 significantly enhanced MMP-9 secretion by iDCs, peaking at 100 ng/ml, which optimally increased iDC migration through a reconstituted basement membrane (Matrigel™) in vitro. The CCL5-enhanced secretion of MMP-9 occurred early (2 h) and was maintained at least for 10 h. A significant increase in MMP-9 mRNA synthesis was detected by reverse transcriptase-polymerase chain reaction, only at 6 h of CCL5 treatment, wh...

Research paper thumbnail of TFPI-2 silencing increases tumour progression and promotes metalloproteinase 1 and 3 induction through tumour-stromal cell interactions

Journal of Cellular and Molecular Medicine, 2011

Tumour progression is a complex multistep process that depends on an evolving crosstalk between c... more Tumour progression is a complex multistep process that depends on an evolving crosstalk between cancer cells and the surrounding stromal tissue. The microenvironment is now recognized as having a pivotal role in promoting cancer initiation, progression and dissemination to form metastases [1]. The invasion process involves extracellular matrix (ECM)-degrading proteases, particularly matrix metalloproteinases (MMPs), that have been shown to be highly expressed and activated in the tumour microenvironment [2], especially in highly aggressive malignant tumours [3, 4]. Activated fibroblasts, the major cell component of the microenvironment, actively contribute to tumour invasiveness by secreting a consistent amount of MMPs at the tumour-stroma interface. Moreover, this MMP synthesis could be increased by the ECM metalloproteinase inducer (EMMPRIN) expressed by cancer cells. Activation of zymogens (pro-MMPs) in the extracellular environment needs proteolytic cleavage of the aminoterminal prodomain [5] that depends on serine proteases, such as trypsin and plasmin, and also involves activated MMPs or membrane-anchored matrix metalloproteinases (MT-MMPs). Apart from tissue inhibitors of metalloproteinases (TIMPs) that are specific regulators of MMP activity, TFPI-2 (tissue factor pathway inhibitor-2), an inhibitor of serine proteases-particularly plasmin-could also regulate the activation of MMPs [6, 7], thus regulating ECM degradation and tumour cell invasion. TFPI-2 silencing increases tumour progression and promotes metalloproteinase 1 and 3 induction through tumour-stromal cell interactions