Christian Larochelle | Université Laval (original) (raw)
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Papers by Christian Larochelle
PLOS One, 2010
Background: The genomic organization of Hox clusters is fundamental for the precise spatio-tempor... more Background: The genomic organization of Hox clusters is fundamental for the precise spatio-temporal regulation and the function of each Hox gene, and hence for correct embryo patterning. Multiple overlapping transcriptional units exist at the Hoxa5 locus reflecting the complexity of Hox clustering: a major form of 1.8 kb corresponding to the two characterized exons of the gene and polyadenylated RNA species of 5.0, 9.5 and 11.0 kb. This transcriptional intricacy raises the question of the involvement of the larger transcripts in Hox function and regulation.
Breast Cancer Research, 2008
Introduction Estrogen and androgen signalling pathways exert opposing influences on the prolifera... more Introduction Estrogen and androgen signalling pathways exert opposing influences on the proliferation of mammary epithelial and hormone-dependent breast cancer cells. We previously reported that plasma concentrations of 1,1-dichloro-2,2-bis(pchlorophenyl)ethylene (p,p'-DDE), the main metabolite of the insecticide DDT (1,1,1-trichloro-2,2-bis [p-chlorophenyl]ethane) and a potent androgen antagonist, were associated with tumor aggressiveness in women diagnosed with breast cancer. We sought to examine the biological plausibility of this association by testing the effect of p,p'-DDE on the proliferation of CAMA-1 cells, a human breast cancer cell line that expresses the estrogen receptor alpha (ERα) and the androgen receptor (AR), in the presence of physiological concentrations of estrogens and androgens in the cell culture medium.
PLOS One, 2010
Background: The genomic organization of Hox clusters is fundamental for the precise spatio-tempor... more Background: The genomic organization of Hox clusters is fundamental for the precise spatio-temporal regulation and the function of each Hox gene, and hence for correct embryo patterning. Multiple overlapping transcriptional units exist at the Hoxa5 locus reflecting the complexity of Hox clustering: a major form of 1.8 kb corresponding to the two characterized exons of the gene and polyadenylated RNA species of 5.0, 9.5 and 11.0 kb. This transcriptional intricacy raises the question of the involvement of the larger transcripts in Hox function and regulation.
Developmental Dynamics, 1999
Genetic analyses have revealed the essential role of the murine Hoxa5 gene for the correct specif... more Genetic analyses have revealed the essential role of the murine Hoxa5 gene for the correct specification of the cervical and upper thoracic region of the skeleton, and for the normal organogenesis and function of the respiratory tract, both structures expressing Hoxa5 during embryogenesis. To understand how the expression domains of the Hoxa5 gene are established during development, we have analyzed the cis-acting control regions mediating Hoxa5 gene expression using a transgenic approach. Four transcripts are derived from the Hoxa5 locus. The shortest and most abundant one displays a specific spatio-temporal profile of expression at earlier stages and in more anterior structures along the embryonic axis than the larger forms. We established that an 11.1 kilobase pair (kb) genomic fragment, extending from position -3.8 kb to +7.3 kb relative to Hoxa5 transcription initiation site, was sufficient to reproduce the temporal expression and substantially reconstitute the spatial pattern of the major Hoxa5 transcript. By deletion analyses, we identified a 2.1 kb fragment located downstream of the Hoxa5 gene that possesses mesodermal enhancer activity. Overall, the findings demonstrate that cis-acting regulatory elements essential for the correct expression of the major Hoxa5 transcript are located both upstream and downstream of the Hoxa5 coding sequences.
Environmental Health Perspectives, 2005
The Lower North Shore region of the St. Lawrence River is home to a fish-eating population that d... more The Lower North Shore region of the St. Lawrence River is home to a fish-eating population that displays an unusually high body burden of several organochlorines, including polychlorinated biphenyls (PCBs) and dioxin-like compounds (DLCs). We measured biomarkers indicative of liver enzyme induction and investigated the relationship with organochlorine body burden in adult volunteers from this population. We determined plasma concentrations of PCBs and chlorinated pesticides by high-resolution gas chromatography (HRGC) with electron capture detection. DLC concentrations were measured by the dioxin-receptor chemically activated luciferase expression (DR-CALUX) assay and in a subset of participants, by HRGC/high-resolution mass spectrometry. We measured cotinine, D-glucaric acid, and porphyrins in morning urine samples and determined liver CYP1A2 activity in vivo using the caffeine breath test. Neither DLC concentrations as measured by the DR-CALUX nor PCB-153 concentrations, the latter representing total PCB exposure, were correlated with biomarkers of effects. Smoking (morning urinary cotinine concentration) was positively related to CYP1A2 activity as measured by the caffeine breath test (p < 0.01). Liver CYP1A2 activity was in turn negatively correlated with PCB-105:PCB-153 and PCB-118:PCB-153 congener ratios (p < 0.05). Hence, despite the relatively high body burden of PCBs and DLCs in this population, only smoking had a significant correlation with biomarkers of hepatic enzyme induction. Our data are consistent with smoking-induced liver CYP1A2 activity altering heme metabolism and increasing the biotransformation of mono-ortho PCB congeners.
Developmental Biology, 1999
The Hox genes cooperate in providing positional information needed for spatial and temporal patte... more The Hox genes cooperate in providing positional information needed for spatial and temporal patterning of the vertebrate body axis. However, the biological mechanisms behind spatial Hox expression are largely unknown. In transgenic mice, gene fusions between Hoxa5 (previously called Hox-1.3) 5 flanking regions and the lacZ reporter gene show tissue-and time-specific expression in the brachial spinal cord in day 11-13 embryos. A 604-bp regulatory region with enhancer properties directs this spatially specific expression. Fine-detail mapping of the enhancer has identified several elements involved in region-specific expression, including an element required for expression in the brachial spinal cord. Factors in embryonic day 12.5 nuclear extracts bind this element in electrophoretic mobility shift assays (EMSA) and protect three regions from DNase digestion. All three sites contain an AAATAA sequence and mutations at these sites reduce or abolish binding. Furthermore, this element binds specific individual embryonic proteins on a protein blot. The binding activity appears as a gradient along the anterior-posterior axis with two-to threefold higher levels observed in extracts from anterior regions than from posterior regions. In parallel with the EMSA, the proteins on the protein blot also show reduced binding to probes with mutations at the AAATAA sites. Most importantly, transgenic mice carrying Hoxa5/lacZ fusions with the three AAATAA sites mutated either do not express the transgene or have altered transgene expression. The brachial spinal cord element and its binding proteins are likely to be involved in spatial expression of Hoxa5 during development.
Chemosphere, 2010
The exposure of Inuit people to polychlorinated biphenyls (PCBs) and chlorinated pesticides has b... more The exposure of Inuit people to polychlorinated biphenyls (PCBs) and chlorinated pesticides has been well characterised but little is known regarding their exposure to dioxin-like compounds, which induce toxic effects through binding to the aryl hydrocarbon receptor (AhR). In order to obtain a global measure of persistent organic pollutants in plasma that interact with this signalling pathway, we used a luciferase reporter gene assay to assess the AhR-mediated transcriptional activity elicited by plasma sample extracts from 874 Inuit adults who were recruited in the course of a prospective epidemiological study conducted in Nunavik (Québec, Canada). Several sociodemographic, anthropometric, dietary and lifestyle variables were considered as possible modulating factors of the AhR-mediated activity in multivariate statistical analyses. The geometric mean AhR-mediated activity expressed as 2,3,7,8-tetachlorodibenzo-p-dioxin equivalents was 8.9 pg g À1 lipids (range: <5-144 pg g À1 lipids). PCB-153 concentration measured by high-resolution gas chromatography-mass spectrometry was moderately correlated to AhR-mediated activity (Pearson's r = 0.53, p < 0.001). Multiple linear regression analyses revealed that age and omega-3 fatty acids in erythrocyte membranes (an index of marine food consumption) were positively associated with plasma AhR-mediated activity (p < 0.001), whereas a negative association was noted with body fat mass (p = 0.037). These results suggest that AhR-mediated transcriptional activity of Inuit plasma extracts is linked to their organochlorine body burden, most likely that of dioxin-like PCBs, polychlorinated dibenzo-p-dioxins and polychlorodibenzofurans. AhR-mediated transcriptional activity measures may prove useful in investigating possible associations between exposure to AhR agonists and adverse health effects in this indigenous population. (T.C.M. Medehouenou), christian.larochelle@inspq.qc.ca (C. Larochelle), pierre.dumas@inspq.qc.ca (P. Dumas), eric.dewailly@crchul.ulaval.ca (É. Dewailly), pierre.ayotte@inspq.qc.ca (P. Ayotte).
Environmental Research, 2011
Organochlorine compounds (OCs) are a group of persistent chemicals that accumulate in fatty tissu... more Organochlorine compounds (OCs) are a group of persistent chemicals that accumulate in fatty tissues with age. Although OCs has been tested individually for their capacity to induce breast cancer cell proliferation, few studies examined the effect of complex mixtures that comprise compounds frequently detected in the serum of women. We constituted such an OC mixture containing 15 different components in environmentally relevant proportions and assessed its proliferative effects in four breast cancer cell lines (MCF-7, T47D, CAMA-1, MDAMB231) and in non-cancerous CV-1 cells. We also determined the capacity of the mixture to modulate cell cycle stage of breast cancer cells and to induce estrogenic and antiandrogenic effects using gene reporter assays. We observed that low concentrations of the mixture (100 Â 10 3 and 50 Â 10 3 dilutions) stimulated the proliferation of MCF-7 cells while higher concentrations (10 Â 10 3 and 5 Â 10 3 dilutions) had the opposite effect. In contrast, the mixture inhibited the proliferation of non-hormone-dependent cell lines. The mixture significantly increased the number of MCF-7 cells entering the S phase, an effect that was blocked by the antiestrogen ICI 182,780. Low concentrations of the mixture also caused an increase in CAMA-1 cell proliferation but only in the presence estradiol and dihydrotestosterone (po 0.05 at the 50 Â 10 3 dilution). DDT analogs and polychlorinated biphenyls all had the capacity to stimulate the proliferation of CAMA-1 cells in the presence of sex steroids. Reporter gene assays further revealed that the mixture and several of its constituents (DDT analogs, aldrin, dieldrin, b-hexachlorocyclohexane, toxaphene) induced estrogenic effects, whereas the mixture and several components (DDT analogs, aldrin, dieldrin and PCBs) inhibited the androgen signaling pathway. Our results indicate that the complex OC mixture increases the proliferation of MCF-7 cells due to its estrogenic potential. The proliferative effect of the mixture on CAMA-1 cells in the presence of sex steroids appears mostly due to the antiandrogenic properties of p,p 0 -DDE, a major constituent of the mixture. Other mixtures of contaminants that include emerging compounds of interest such as brominated flame retardants and perfluoroalkyl compounds should be tested for their capacity to induce breast cancer cell proliferation.
PLOS One, 2010
Background: The genomic organization of Hox clusters is fundamental for the precise spatio-tempor... more Background: The genomic organization of Hox clusters is fundamental for the precise spatio-temporal regulation and the function of each Hox gene, and hence for correct embryo patterning. Multiple overlapping transcriptional units exist at the Hoxa5 locus reflecting the complexity of Hox clustering: a major form of 1.8 kb corresponding to the two characterized exons of the gene and polyadenylated RNA species of 5.0, 9.5 and 11.0 kb. This transcriptional intricacy raises the question of the involvement of the larger transcripts in Hox function and regulation.
Breast Cancer Research, 2008
Introduction Estrogen and androgen signalling pathways exert opposing influences on the prolifera... more Introduction Estrogen and androgen signalling pathways exert opposing influences on the proliferation of mammary epithelial and hormone-dependent breast cancer cells. We previously reported that plasma concentrations of 1,1-dichloro-2,2-bis(pchlorophenyl)ethylene (p,p'-DDE), the main metabolite of the insecticide DDT (1,1,1-trichloro-2,2-bis [p-chlorophenyl]ethane) and a potent androgen antagonist, were associated with tumor aggressiveness in women diagnosed with breast cancer. We sought to examine the biological plausibility of this association by testing the effect of p,p'-DDE on the proliferation of CAMA-1 cells, a human breast cancer cell line that expresses the estrogen receptor alpha (ERα) and the androgen receptor (AR), in the presence of physiological concentrations of estrogens and androgens in the cell culture medium.
PLOS One, 2010
Background: The genomic organization of Hox clusters is fundamental for the precise spatio-tempor... more Background: The genomic organization of Hox clusters is fundamental for the precise spatio-temporal regulation and the function of each Hox gene, and hence for correct embryo patterning. Multiple overlapping transcriptional units exist at the Hoxa5 locus reflecting the complexity of Hox clustering: a major form of 1.8 kb corresponding to the two characterized exons of the gene and polyadenylated RNA species of 5.0, 9.5 and 11.0 kb. This transcriptional intricacy raises the question of the involvement of the larger transcripts in Hox function and regulation.
Developmental Dynamics, 1999
Genetic analyses have revealed the essential role of the murine Hoxa5 gene for the correct specif... more Genetic analyses have revealed the essential role of the murine Hoxa5 gene for the correct specification of the cervical and upper thoracic region of the skeleton, and for the normal organogenesis and function of the respiratory tract, both structures expressing Hoxa5 during embryogenesis. To understand how the expression domains of the Hoxa5 gene are established during development, we have analyzed the cis-acting control regions mediating Hoxa5 gene expression using a transgenic approach. Four transcripts are derived from the Hoxa5 locus. The shortest and most abundant one displays a specific spatio-temporal profile of expression at earlier stages and in more anterior structures along the embryonic axis than the larger forms. We established that an 11.1 kilobase pair (kb) genomic fragment, extending from position -3.8 kb to +7.3 kb relative to Hoxa5 transcription initiation site, was sufficient to reproduce the temporal expression and substantially reconstitute the spatial pattern of the major Hoxa5 transcript. By deletion analyses, we identified a 2.1 kb fragment located downstream of the Hoxa5 gene that possesses mesodermal enhancer activity. Overall, the findings demonstrate that cis-acting regulatory elements essential for the correct expression of the major Hoxa5 transcript are located both upstream and downstream of the Hoxa5 coding sequences.
Environmental Health Perspectives, 2005
The Lower North Shore region of the St. Lawrence River is home to a fish-eating population that d... more The Lower North Shore region of the St. Lawrence River is home to a fish-eating population that displays an unusually high body burden of several organochlorines, including polychlorinated biphenyls (PCBs) and dioxin-like compounds (DLCs). We measured biomarkers indicative of liver enzyme induction and investigated the relationship with organochlorine body burden in adult volunteers from this population. We determined plasma concentrations of PCBs and chlorinated pesticides by high-resolution gas chromatography (HRGC) with electron capture detection. DLC concentrations were measured by the dioxin-receptor chemically activated luciferase expression (DR-CALUX) assay and in a subset of participants, by HRGC/high-resolution mass spectrometry. We measured cotinine, D-glucaric acid, and porphyrins in morning urine samples and determined liver CYP1A2 activity in vivo using the caffeine breath test. Neither DLC concentrations as measured by the DR-CALUX nor PCB-153 concentrations, the latter representing total PCB exposure, were correlated with biomarkers of effects. Smoking (morning urinary cotinine concentration) was positively related to CYP1A2 activity as measured by the caffeine breath test (p < 0.01). Liver CYP1A2 activity was in turn negatively correlated with PCB-105:PCB-153 and PCB-118:PCB-153 congener ratios (p < 0.05). Hence, despite the relatively high body burden of PCBs and DLCs in this population, only smoking had a significant correlation with biomarkers of hepatic enzyme induction. Our data are consistent with smoking-induced liver CYP1A2 activity altering heme metabolism and increasing the biotransformation of mono-ortho PCB congeners.
Developmental Biology, 1999
The Hox genes cooperate in providing positional information needed for spatial and temporal patte... more The Hox genes cooperate in providing positional information needed for spatial and temporal patterning of the vertebrate body axis. However, the biological mechanisms behind spatial Hox expression are largely unknown. In transgenic mice, gene fusions between Hoxa5 (previously called Hox-1.3) 5 flanking regions and the lacZ reporter gene show tissue-and time-specific expression in the brachial spinal cord in day 11-13 embryos. A 604-bp regulatory region with enhancer properties directs this spatially specific expression. Fine-detail mapping of the enhancer has identified several elements involved in region-specific expression, including an element required for expression in the brachial spinal cord. Factors in embryonic day 12.5 nuclear extracts bind this element in electrophoretic mobility shift assays (EMSA) and protect three regions from DNase digestion. All three sites contain an AAATAA sequence and mutations at these sites reduce or abolish binding. Furthermore, this element binds specific individual embryonic proteins on a protein blot. The binding activity appears as a gradient along the anterior-posterior axis with two-to threefold higher levels observed in extracts from anterior regions than from posterior regions. In parallel with the EMSA, the proteins on the protein blot also show reduced binding to probes with mutations at the AAATAA sites. Most importantly, transgenic mice carrying Hoxa5/lacZ fusions with the three AAATAA sites mutated either do not express the transgene or have altered transgene expression. The brachial spinal cord element and its binding proteins are likely to be involved in spatial expression of Hoxa5 during development.
Chemosphere, 2010
The exposure of Inuit people to polychlorinated biphenyls (PCBs) and chlorinated pesticides has b... more The exposure of Inuit people to polychlorinated biphenyls (PCBs) and chlorinated pesticides has been well characterised but little is known regarding their exposure to dioxin-like compounds, which induce toxic effects through binding to the aryl hydrocarbon receptor (AhR). In order to obtain a global measure of persistent organic pollutants in plasma that interact with this signalling pathway, we used a luciferase reporter gene assay to assess the AhR-mediated transcriptional activity elicited by plasma sample extracts from 874 Inuit adults who were recruited in the course of a prospective epidemiological study conducted in Nunavik (Québec, Canada). Several sociodemographic, anthropometric, dietary and lifestyle variables were considered as possible modulating factors of the AhR-mediated activity in multivariate statistical analyses. The geometric mean AhR-mediated activity expressed as 2,3,7,8-tetachlorodibenzo-p-dioxin equivalents was 8.9 pg g À1 lipids (range: <5-144 pg g À1 lipids). PCB-153 concentration measured by high-resolution gas chromatography-mass spectrometry was moderately correlated to AhR-mediated activity (Pearson's r = 0.53, p < 0.001). Multiple linear regression analyses revealed that age and omega-3 fatty acids in erythrocyte membranes (an index of marine food consumption) were positively associated with plasma AhR-mediated activity (p < 0.001), whereas a negative association was noted with body fat mass (p = 0.037). These results suggest that AhR-mediated transcriptional activity of Inuit plasma extracts is linked to their organochlorine body burden, most likely that of dioxin-like PCBs, polychlorinated dibenzo-p-dioxins and polychlorodibenzofurans. AhR-mediated transcriptional activity measures may prove useful in investigating possible associations between exposure to AhR agonists and adverse health effects in this indigenous population. (T.C.M. Medehouenou), christian.larochelle@inspq.qc.ca (C. Larochelle), pierre.dumas@inspq.qc.ca (P. Dumas), eric.dewailly@crchul.ulaval.ca (É. Dewailly), pierre.ayotte@inspq.qc.ca (P. Ayotte).
Environmental Research, 2011
Organochlorine compounds (OCs) are a group of persistent chemicals that accumulate in fatty tissu... more Organochlorine compounds (OCs) are a group of persistent chemicals that accumulate in fatty tissues with age. Although OCs has been tested individually for their capacity to induce breast cancer cell proliferation, few studies examined the effect of complex mixtures that comprise compounds frequently detected in the serum of women. We constituted such an OC mixture containing 15 different components in environmentally relevant proportions and assessed its proliferative effects in four breast cancer cell lines (MCF-7, T47D, CAMA-1, MDAMB231) and in non-cancerous CV-1 cells. We also determined the capacity of the mixture to modulate cell cycle stage of breast cancer cells and to induce estrogenic and antiandrogenic effects using gene reporter assays. We observed that low concentrations of the mixture (100 Â 10 3 and 50 Â 10 3 dilutions) stimulated the proliferation of MCF-7 cells while higher concentrations (10 Â 10 3 and 5 Â 10 3 dilutions) had the opposite effect. In contrast, the mixture inhibited the proliferation of non-hormone-dependent cell lines. The mixture significantly increased the number of MCF-7 cells entering the S phase, an effect that was blocked by the antiestrogen ICI 182,780. Low concentrations of the mixture also caused an increase in CAMA-1 cell proliferation but only in the presence estradiol and dihydrotestosterone (po 0.05 at the 50 Â 10 3 dilution). DDT analogs and polychlorinated biphenyls all had the capacity to stimulate the proliferation of CAMA-1 cells in the presence of sex steroids. Reporter gene assays further revealed that the mixture and several of its constituents (DDT analogs, aldrin, dieldrin, b-hexachlorocyclohexane, toxaphene) induced estrogenic effects, whereas the mixture and several components (DDT analogs, aldrin, dieldrin and PCBs) inhibited the androgen signaling pathway. Our results indicate that the complex OC mixture increases the proliferation of MCF-7 cells due to its estrogenic potential. The proliferative effect of the mixture on CAMA-1 cells in the presence of sex steroids appears mostly due to the antiandrogenic properties of p,p 0 -DDE, a major constituent of the mixture. Other mixtures of contaminants that include emerging compounds of interest such as brominated flame retardants and perfluoroalkyl compounds should be tested for their capacity to induce breast cancer cell proliferation.