Role of estrogen receptor alpha in human cervical cancer-associated fibroblasts: a transcriptomic study
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Cancer-Associated Fibroblasts (CAFs) are crucial in genesis and progression of tumors; however, cervical CAFs (C-CAFs) are not well characterized. Estradiol (E2) has been implicated as a cofactor in human papillomavirus (HPV)-mediated cervical cancer (CxCa), both in animal models and in women using oral contraceptives; however, the exact role of the hormone is unclear. Human C-CAFs have recently been shown to express estrogen receptor alpha (ER-α). We investigated gene expression patterns in ex vivo cultured early and late stage C-CAFs in the context of E2. CAFs were isolated from four patients with early and two patients with late stage CxCa. ER-α expression in CxCa tissues was localized to stromal fibroblast-like cells and confirmed in ex vivo cultured C-CAFs. Two ER antagonists (ICI 182,780 and Methyl Piperidino Pyrazole) were used to unravel ER signaling in CAFs. Microarray technology was used for expression profiling and validated by quantitative reverse transcription PCR. The transcriptomes of C-CAFs across stages indicated their activated state. C-CAFs had gene expression patterns associated with both pro-tumorigenic and pro-inflammatory signaling. Late-stage C-CAFs compared to those of early stage appeared to be more actively metabolizing and cycling but expressed fewer genes related to immune function. We report differential expression profiles between C-CAFs: early vs. late stage and in the presence of ER antagonists. Both ER antagonists seemed to modulate C-CAF function by down regulating genes associated with cell cycle and metabolism, affecting angiogenesis and cancer progression. This study characterized C-CAFs from early and late stage disease, and experiments with ER inhibitors emphasized the probable importance of canonical ER-α signaling. Interfering with paracrine signaling through fibroblast ER-α is worth exploiting as a targeted therapy in CxCa management.
KeywordsCervical cancer Cancer-Associated Fibroblasts Estrogen Receptor Alpha Estrogen Receptor antagonists ICI 182,780 MPP-Methyl Piperidino Pyrazole
The authors wish to thank Swathi U. Lekshmi (Senior Research Fellow, Department of Microbiology, KMIO, Bangalore) for her help with immunohistochemistry, Anita Mahadevan (Additional Professor, Department of Neuropathology, National Institute of Mental Health and Neurosciences) for her help in microphotography, Annapoorni Rangarajan (Associate Professor, MRDG, IISc, Bangalore) for her help during the initial stages of CAF isolation, Mahua Sinha (Assistant Professor, Department of Microbiology, KMIO) and Reeta Mani (Associate Professor, Neurovirology, NIMHANS, Bangalore) for editing the manuscript, Neta Erez (Department of Pathology, Sackler School of Medicine, Tel Aviv University, Tel Aviv) for kindly reviewing the manuscript, and Harsha Gowda (Faculty Scientist, Institute of Bioinformatics, Bangalore) for useful discussions and for improving the manuscript. The authors gratefully acknowledge the financial support of RGCB, Thiruvananthapuram, for part funding of the expenditure towards microarray. Financial support from the Indian Council of Medical Research (ICMR), India (No. 5/13/127/2011/NCD-III), and the Council of Scientific and Industrial Research (CSIR), India, for Senior Research Fellowship to MKM (09/999/(0001)/2009-EMR-I) is also acknowledged.
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Conflicts of interest
Financial competing interests
The research and development division of a commercial organization (Shodhaka LS Pvt. Ltd.) was involved in the research work of SD, AKB, and AKK. However, the participation was independent of the commercial interests of the company. There are no financial competing interests in conducting or reporting this work.
All procedures performed in studies involving human participants were in accordance with the Institutional Ethics Committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.
Written informed consent was obtained from all individual participants included in the study.
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