Daniela Costea - Academia.edu (original) (raw)

Papers by Daniela Costea

Oral Oncology Supplement, 2005

Introduction: Basement membrane proteins are considered to play a siginficant role in epithelial ... more Introduction: Basement membrane proteins are considered to play a siginficant role in epithelial tumour progression Our objectave an this study was to Inveshgate the express:on of basement membrane proteins an m wtro orgas~otyp:c cultures of normal, early neoplashc and estabhshed neoplashc oral mucosa, and to determine their role in tumour progression. Materials and Methods: Organotyp~c culture models of normal, early neoplastic and estabhshed neoplastic human oral mucosa were constructed by growing ( 1 ) prinlary normal human oral keratinocytes, (2) early neoplashc truman oral keratinocytes (DOK cell line) and (3) estabhshed neoplashc human oral keratinocytes (PA-CA/PJ 15 cell line) on top of Collagen type I matrices In the presence or absence of prHnary normal human oral fibroblasts. Inmmnohistochemlcal staining was performed for detechon of Collagen IV and Lan~mm V with monoclonal anhboches, and for F:bronectln by using a polyclonal anhbody. Results: Collagen IV was detected at the ep:thehal-matrix interface of normal, early neoplashc and estabhshed neoplastic models developed w~th fibroblast-contammg Collagen type I matrix, but only in the estabhshed neoplastic model constructed wlth CoUagen type I matrix without fibroblasts. Lanmm: V was also detected at the ep:thella1-matrax mterl~ace of all the three models wath fibroblast-contaming collagen matrices. Fabronectin was present at the epathehal-matrix Intert~ace as well as In the matrax adjacent to fibroblasts In all fl~ree models constructed with fibroblast-containing collagen matrices. The Flbronectin component an the collagen I matrax increased with the degree ofkeratanocyte transformatwn. There was a decrease an anvaslveness of neoplastac cells when collagen IV antibody was added to the culture medaun:. Matrix coating with Collagen IV dad not promote neoplastic cell Invasaon Conelusion: Basement membrane proteins are expressed In an vitro models of normal, early neoplashc and estabhshed neoplashc oral mucosa, and the:r deposition seems to be fibroblast-dependent. Collagen IV and F:bronectin may play a role In ~:our cell Invaslveness.

Cancer Research, 2013

Heterogeneity of carcinoma-associated fibroblasts (CAF) has long been recognized, but the functio... more Heterogeneity of carcinoma-associated fibroblasts (CAF) has long been recognized, but the functional significance remains poorly understood. Here, we report the distinction of two CAF subtypes in oral squamous cell carcinoma (OSCC) that have differential tumor-promoting capability, one with a transcriptome and secretome closer to normal fibroblasts (CAF-N) and the other with a more divergent expression pattern (CAF-D). Both subtypes supported higher tumor incidence in nonobese diabetic/severe combined immunodeficient (NOD/SCID) Ilγ2(null) mice and deeper invasion of malignant keratinocytes than normal or dysplasia-associated fibroblasts, but CAF-N was more efficient than CAF-D in enhancing tumor incidence. CAF-N included more intrinsically motile fibroblasts maintained by high autocrine production of hyaluronan. Inhibiting CAF-N migration by blocking hyaluronan synthesis or chain elongation impaired invasion of adjacent OSCC cells, pinpointing fibroblast motility as an essential mechanism in this process. In contrast, CAF-D harbored fewer motile fibroblasts but synthesized higher TGF-β1 levels. TGF-β1 did not stimulate CAF-D migration but enhanced invasion and expression of epithelial-mesenchymal transition (EMT) markers in malignant keratinocytes. Inhibiting TGF-β1 in three-dimensional cultures containing CAF-D impaired keratinocyte invasion, suggesting TGF-β1-induced EMT mediates CAF-D-induced carcinoma cell invasion. TGF-β1-pretreated normal fibroblasts also induced invasive properties in transformed oral keratinocytes, indicating that TGF-β1-synthesizing fibroblasts, as well as hyaluronan-synthesizing fibroblasts, are critical for carcinoma invasion. Taken together, these results discern two subtypes of CAF that promote OSCC cell invasion via different mechanisms.

Archives of Oral Biology, 2009

Cancer Fibroblasts a b s t r a c t Background: Although basement membrane was traditionally consi... more Cancer Fibroblasts a b s t r a c t Background: Although basement membrane was traditionally considered an inert barrier that tumour cells had to cross before invasion into the surrounding stroma, recent studies suggest that basement membrane components are not only degraded during tumour progression, but also newly synthesised at the invasive front.

The American Journal of Pathology, 2006

This study tests the hypothesis that invasion of partially transformed keratinocytes is initiated... more This study tests the hypothesis that invasion of partially transformed keratinocytes is initiated by diffusible, proinvasive signals provided by species-specific fibroblasts. In vitro organotypic cultures of neoplastic human oral mucosa were constructed by growing a partially transformed, nontumorigenic keratinocytic cell line isolated from a dysplastic human oral lesion (DOK-ECACC94122104) on top of various types of connective tissue equivalents. Cultured tissues were analyzed by histomorphometry (depth and area of invasion: D inv , A inv ) and immunohistochemistry. Presence of human fibroblasts in the matrix induced a local invasion of DOK (D inv ‫؍‬ 95.6 ؎ 7.1 m, A inv ‫؍‬ 45.8 ؎ 3.5%). Minimal invasion (P < 0.05) was observed when DOK grew on simple collagen matrix (D inv ‫؍‬ 14.1 ؎ 2.1 m, A inv ‫؍‬ 3.7 ؎ 0.8%) or matrices containing fibroblasts from mouse (D inv ‫؍‬ 11.5 ؎ 4.0 m, A inv ‫؍‬ 4.3 ؎ 1.0%) or rat (D inv ‫؍‬ 15.6 ؎ 1.2 m, A inv ‫؍‬ 6.1 ؎ 0.5%). In these cultures, local invasion could be induced by the presence of human fibroblasts in a bottom layer of the collagen matrix (P < 0.05) or by conditioned medium from organotypic cultures of DOK on human fibroblast-containing matrix (P < 0.05) but not by conditioned medium from human fibroblast monocultures (P > 0.05). Deposition of human collagen IV was observed at epithelialmatrix interface only when DOK behaved invasively. In conclusion, invasion of partially transformed oral keratinocytes was triggered by keratinocyte-induced fibroblast-derived diffusible factor(s) in a species-specific manner and associated with de novo synthesis of collagen

The American Journal of Pathology, Jun 1, 2006

Oral Oncology Supplement, 2005

The International Journal of Biochemistry & Cell Biology, 2016

Vimentin expression correlates well with migratory and invasive potential of the carcinoma cells.... more Vimentin expression correlates well with migratory and invasive potential of the carcinoma cells. The molecular mechanism by which vimentin regulates cell motility is not yet clear. Here, we addressed this issue by depleting vimentin in oral squamous cell carcinoma derived cell line. Vimentin knockdown cells showed enhanced adhesion and spreading to laminin-5. However, we found that they were less invasive as compared to the vector control cells. In addition, signaling associated with adhesion behavior of the cell was increased in vimentin knockdown clones. These findings suggest that the normal function of β4 integrin as mechanical adhesive device is enhanced upon vimentin downregulation. As a proof of principle, the compromised invasive potential of vimentin depleted cells could be rescued upon blocking with β4 integrin adhesion-blocking (ASC-8) antibody or downregulation of β4 integrin in vimentin knockdown background. Interestingly, plectin which associates with α6β4 integrin in the hemidesmosomes, was also found to be upregulated in vimentin knockdown clones. Furthermore, experiments on lysosome and proteasome inhibition revealed that perhaps vimentin regulates the turnover of β4 integrin and plectin. Moreover, an inverse association was observed between vimentin expression and β4 integrin in oral squamous cell carcinoma (OSCC). Collectively, our results show a novel role of vimentin in modulating cell motility by destabilizing β4 integrin-mediated adhesive interactions. Further, vimentin-β4 integrin together may prove to be useful markers for prognostication of human oral cancer.

Advanced healthcare materials, Jan 8, 2016

The aim is to evaluate the effect of modifying poly[(l-lactide)-co-(ε-caprolactone)] scaffolds (P... more The aim is to evaluate the effect of modifying poly[(l-lactide)-co-(ε-caprolactone)] scaffolds (PLCL) with nanodiamonds (nDP) or with nDP+physisorbed BMP-2 (nDP+BMP-2) on in vivo host tissue response and degradation. The scaffolds are implanted subcutaneously in Balb/c mice and retrieved after 1, 8, and 27 weeks. Molecular weight analysis shows that modified scaffolds degrade faster than the unmodified. Gene analysis at week 1 shows highest expression of proinflammatory markers around nDP scaffolds; although the presence of inflammatory cells and foreign body giant cells is more prominent around the PLCL. Tissue regeneration markers are highly expressed in the nDP+BMP-2 scaffolds at week 8. A fibrous capsule is detectable by week 8, thinnest around nDP scaffolds and at week 27 thickest around PLCL scaffolds. mRNA levels of ALP, COL1α2, and ANGPT1 are significantly upregulating in the nDP+BMP-2 scaffolds at week 1 with ectopic bone seen at week 8. Even when almost 90% of the scaffold...

EBioMedicine, 2016

Cancer stem cells (CSCs) drive tumour spread and therapeutic resistance, and can undergo epitheli... more Cancer stem cells (CSCs) drive tumour spread and therapeutic resistance, and can undergo epithelial-to-mesenchymal transition (EMT) and mesenchymal-to-epithelial transition (MET) to switch between epithelial and post-EMT sub-populations. Examining oral squamous cell carcinoma (OSCC), we now show that increased phenotypic plasticity, the ability to undergo EMT/MET, underlies increased CSC therapeutic resistance within both the epithelial and post-EMT sub-populations. The post-EMT CSCs that possess plasticity exhibit particularly enhanced therapeutic resistance and are defined by a CD44(high)EpCAM(low/-) CD24(+) cell surface marker profile. Treatment with TGFβ and retinoic acid (RA) enabled enrichment of this sub-population for therapeutic testing, through which the endoplasmic reticulum (ER) stressor and autophagy inhibitor Thapsigargin was shown to selectively target these cells. Demonstration of the link between phenotypic plasticity and therapeutic resistance, and development of an in vitro method for enrichment of a highly resistant CSC sub-population, provides an opportunity for the development of improved chemotherapeutic agents that can eliminate CSCs.

BioMed research international, 2015

Oral cancer, represented mainly by oral squamous cell carcinoma (OSCC), is the eighth most common... more Oral cancer, represented mainly by oral squamous cell carcinoma (OSCC), is the eighth most common type of human cancer worldwide. The number of new OSCC cases is increasing worldwide, especially in the low-income countries, and the prognosis remains poor in spite of recent advances in the diagnostic and therapeutic modalities. MicroRNAs (miRNAs), 18-25 nucleotides long noncoding RNA molecules, have recently gained significant attention as potential regulators and biomarkers for carcinogenesis. Recent data show that several miRNAs are deregulated in OSCC, and they have either a tumor suppressive or an oncogenic role in oral carcinogenesis. This review summarizes current knowledge on the role of miRNAs as tumor promotors or tumor suppressors in OSCC development and discusses their potential value as diagnostic and prognostic markers in OSCC.

BMC Cancer, 2015

Altered expression of S100A16 has been reported in human cancers, but its biological role in tumo... more Altered expression of S100A16 has been reported in human cancers, but its biological role in tumorigenesis is not fully understood. This study aimed to investigate the clinical significance and functional role of S100A16 in oral squamous cell carcinoma (OSCC) suppression. S100A16 mRNA and/or protein levels were examined by quantitative RT-PCR and immunohistochemistry in whole- and laser microdissected-specimens of normal human oral mucosa (NHOM, n = 65), oral dysplastic lesions (ODL, n = 21), OSCCs (n = 132) and positive cervical nodes (n = 17). S100A16 protein expression in OSCC was examined for correlations with clinicopathological variables and patient survival. S100A16 was over-expressed and knocked-down in OSCC-derived (CaLH3 and H357) cells by employing retroviral constructs to investigate its effects on cell proliferation, sphere formation and three dimensional (3D)-organotypic invasive abilities in vitro and tumorigenesis in a mouse xenograft model. Both S100A16 mRNA and protein levels were found to be progressively down-regulated from NHOM to ODL and OSCC. Low S100A16 protein levels in OSCC significantly correlated with reduced 10-year overall survival and poor tumor differentiation. Analysis of two external OSCC microarray datasets showed a positive correlation between the mRNA expression levels of S100A16 and keratinocyte differentiation markers. CaLH3 and H357 cell fractions enriched for differentiated cells either by lack of adherence to collagen IV or FACS sorting for low p75NTR expression expressed significantly higher S100A16 mRNA levels than the subpopulations enriched for less differentiated cells. Corroborating these findings, retroviral mediated S100A16 over-expression and knock-down in CaLH3 and H357 cells led to respective up- and down-regulation of differentiation markers. In vitro functional studies showed significant reduction in cell proliferation, sphere formation and 3D-invasive abilities of CaLH3 and H357 cells upon S100A16 over-expression. These functional effects were associated with concomitant down-regulation of self-renewal (Bmi-1 and Oct 4A) and invasion related (MMP1 and MMP9) molecules. S100A16 over-expression also suppressed tumorigenesis of H357 cells in a mouse xenograft model and the resulting tumor xenografts displayed features/expression of increased differentiation and reduced proliferation/self-renewal. These results indicate that S100A16 is a differentiation promoting protein and might function as a tumor suppressor in OSCC.

Objective: S100A16 is a recent addition to the S100 protein family. Differential expression of th... more Objective: S100A16 is a recent addition to the S100 protein family. Differential expression of this protein has been reported in many human malignancies. However, its precise biological roles in tumorigenesis are currently unknown. This study aimed to examine the expression pattern and functional role of S100A16 in differentiation of oral squamous cell carcinoma (OSCC). Method: S100A16 mRNA and protein levels were examined in OSCC and normal human oral mucosa (NHOM) respectively by qRT-PCR and immunohistochemistry. Employing retroviral mediated over-expression of S100A16 in oral cancer cell-lines, functional roles of S100A16 was investigated by in vitro and in vivo assays and subsequent molecular analyses. Result: S100A16 expression was found to be down-regulated in OSCCs as compared to NHOMs both at the mRNA and protein levels. Poorly differentiated OSCCs were found to be correlated with low S100A16 mRNA and protein levels. Over-expression of S100A16 was associated with reduced pro...

Objective: The aim of this study is to develop a subcutaneous model in NOD/SCID IL2 R gamma null ... more Objective: The aim of this study is to develop a subcutaneous model in NOD/SCID IL2 R gamma null (NOG) mice to evaluate the tumorigenic potential of functionalized/bioactive scaffolds designed for the use in bone tissue engineering. Method: Dysplastic (precancerous) oral keratinocytes (DOK cell line) known not to be tumorigenic in NUDE mice were injected subcutaneously in NOG mice alone (n=6) or together with carcinoma associated fibroblasts (CAF) (n=12) in order to develop an experimental model of microenvironmentally-induced carcinogenesis. DOK were then transfected to express luciferase such that bioluminescence in vivo imaging of cells/scaffold construct could be carried out in parallel with classical morphometric analysis. They were then cultured alone (n=6) or together with CAF (n=6) on poly(L-lactide-co-e-caprolactone) scaffolds and implanted subcutaneously in NOG mice to optimize the model for future use in screening of various scaffolds intended for bone regeneration. Resul...

British journal of cancer, 2004

Khat chewing is a widespread habit that has a deep-rooted sociocultural tradition in Africa and t... more Khat chewing is a widespread habit that has a deep-rooted sociocultural tradition in Africa and the Middle East. The biological effects of khat are inadequately investigated and controversial. For the first time, we show that an organic extract of khat induces a selective type of cell death having all morphological and biochemical features of apoptotic cell death. Khat extract was shown to contain the major alkaloid compounds cathinone and cathine. The compounds alone and in combination also induced apoptosis. Khat-induced apoptosis occurred synchronously in various human cell lines (HL-60, NB4, Jurkat) within 8 h of exposure. It was partially reversed after removal of khat and the effect was dependent on de novo protein synthesis, as demonstrated by cotreatment with cycloheximide. The cell death was blocked by the pan-caspase inhibitor Z-VAD-fmk, and also by submicromolar concentrations of Z-YVAD-fmk and Z-IETD-fmk, inhibitors of caspase-1 and -8, respectively. The 50% inhibition c...

Journal of Translational Medicine, 2014

Although several studies suggest that stromal fibroblasts mediate treatment resistance in several... more Although several studies suggest that stromal fibroblasts mediate treatment resistance in several cancer types, little is known about how tumor-associated astrocytes modulate the treatment response in brain tumors. Since traditionally used metabolic assays do not distinguish metabolic activity between stromal and tumor cells, and since 2-dimensional co-culture system does not recreate the formidable complexity of the microenvironment within 3-dimensional structures such as solid tumor tissue, we instead established a glioblastoma (GBM) cell-specific bioluminescent assay for direct measurements of tumor cell viability in the treatment of clinical relevant drugs. Methods: Using lentiviral transfection, we established a panel of human GBM cell lines constitutively expressing a fusion transgene encoding luciferase and the enhanced green fluorescence protein (eGFP). We then initiated co-cultures with immortalized astrocytes, TNC-1, and the eGFP/Luc GBM cell lines. Next, we treated all eGFP/Luc GBM cell lines with Temozolomide (TMZ) or Doxorubicin, comparing co-cultures of glioblastoma (GBM) cells and TNC-1 astrocytes with mono-cultures of eGFP/Luc GBM cells. Cell viability was quantitated by measuring the luciferase expression.

PLoS ONE, 2013

Tumor microenvironment (TME) is an active player in carcinogenesis and changes in its composition... more Tumor microenvironment (TME) is an active player in carcinogenesis and changes in its composition modify cancer growth. Carcinoma-associated fibroblasts, bone marrow-derived multipotent mesenchymal stem cells (BMMSCs), and inflammatory cells can all affect the composition of TME leading to changes in proliferation, invasion and metastasis formation of carcinoma cells. In this study, we confirmed an interaction between BMMSCs and oral tongue squamous cell carcinoma (OTSCC) cells by analyzing the invasion progression and gene expression pattern. In a 3-dimensional myoma organotypic invasion model the presence of BMMSCs inhibited the proliferation but increased the invasion of OTSCC cells. Furthermore, the signals originating from OTSCC cells up-regulated the expression of inflammatory chemokines by BMMSCs, whereas BMMSC products induced the expression of known invasion linked molecules by carcinoma cells. Particularly, after the cell-cell interactions, the chemokine CCL5 was abundantly secreted from BMMSCs and a function blocking antibody against CCL5 inhibited BMMSC enhanced cancer invasion area. However, CCL5 blocking antibody did not inhibit the depth of invasion. Additionally, after exposure to BMMSCs, the expression of type I collagen mRNA in OTSCC cells was markedly up-regulated. Interestingly, also high expression of type I collagen N-terminal propeptide (PINP) in vivo correlated with the cancerspecific mortality of OTSCC patients, whereas there was no association between cancer tissue CCL5 levels and the clinical parameters. In conclusion, our results suggest that the interaction between BMMSC and carcinoma cells induce cytokine and matrix molecule expression, of which high level of type I collagen production correlates with the prognosis of OTSCC patients.

PLoS ONE, 2013

Both S100A14 and S100A16 are members of the multifunctional S100 protein family. Formation of hom... more Both S100A14 and S100A16 are members of the multifunctional S100 protein family. Formation of homo/ heterodimers is considered to be one of the major mechanisms for S100 proteins to execute their diverse cellular functions. By employing a classical Yeast two hybrid (Y-2 H) screen, we identified S100A16 as the single interaction partner of S100A14. This interaction was verified by co-immunoprecipitation, double indirect immunofluorescence and double immunostaining in specimens of oral squamous cell carcinoma and normal oral mucosa. The functional significance of this interaction was examined by employing retroviral mediated over-expression and knock-down of these proteins in several cancer cell-lines. Over-expression and knock-down of S100A14 led to concomitant up-and down-regulation of S100A16 protein in the cell-lines examined. However, there was no up-regulation of S100A16 mRNA upon S100A14 over-expression, indicating that modulation of S100A16 expression was not due to enhanced transcriptional activity but possibly by post-transcriptional regulation. In contrary, over-expression of S100A16 was associated neither with the up-regulation of S100A14 mRNA nor its protein, suggesting a unidirectional regulation between S100A14 and S100A16. Cellular treatment with protein synthesis inhibitor cycloheximide demonstrated a time-dependent intracellular degradation of both S100A16 and S100A14 proteins. Additionally, regulation of S100A16 and S100A14 degradation was found to be independent of the classical proteasomal and lysosomal pathways of protein degradation. Further studies will therefore be necessary to understand the functional significance of this interaction and the mechanisms on how S100A14 is involved in the regulation of S100A16 expression.

Journal of Oral Pathology & Medicine, 2013

There is now substantial evidence that only a subpopulation of cells in solid cancers is able to ... more There is now substantial evidence that only a subpopulation of cells in solid cancers is able to sustain tumour growth and to re-initiate new tumours. Various cancers and cancer-derived cell lines, including head and neck squamous cell carcinomas (HNSCC), have a subpopulation of cancer stem cells (CSCs), marked by high levels of expression of the CD44 adhesion molecule. However, it has been unclear whether, in addition to acting as a marker, CD44 has functions that directly influence stem cell properties. The aim of this study was to investigate the role of CD44 in the maintenance of the CSC population in HNSCC cell lines. CD44 was down-regulated either by treating cultures with 1 mM sodium butyrate or by the more specific method of knockdown with siRNAs directed against CD44. Changes in CD44 expression levels were assessed at the mRNA and protein levels, and the effects of CD44 down-regulation on cell proliferation and on the fate of the CSC subpopulations were assessed. Reduced CD44 expression resulted in a decreased rate of population expansion, both initially and on repassage, and there was an alteration in colony morphologies indicative of stem cell loss. Down-regulation of CD44 also led to reduced expression of Oct4A, an alternative marker of CSCs. The results suggest that CD44 has a functional role in maintaining stem cell properties in HNSCC cell lines and provides support for the concept that therapies targeting CD44, or its related signalling pathways, may allow development of more efficient treatment strategies.

Oral Oncology Supplement, 2005

Cancer Research, 2013

Archives of Oral Biology, 2009

The American Journal of Pathology, 2006

The American Journal of Pathology, Jun 1, 2006

Oral Oncology Supplement, 2005

The International Journal of Biochemistry & Cell Biology, 2016

Advanced healthcare materials, Jan 8, 2016

EBioMedicine, 2016

BioMed research international, 2015

BMC Cancer, 2015

British journal of cancer, 2004

Journal of Translational Medicine, 2014

PLoS ONE, 2013

Journal of Oral Pathology & Medicine, 2013