K Bahar-Shany1, H Brand2, S Sapoznik1, J Jacob-Hirsch1, Y Yung3, J Korach4, T Perri4, Y Cohen5, A Hourvitz6, K Levanon7. 1. Sheba Cancer Research Center, Chaim Sheba Medical Center, Ramat Gan 52621, Israel. 2. Sheba Cancer Research Center, Chaim Sheba Medical Center, Ramat Gan 52621, Israel; Sackler Faculty of Medicine, Tel-Aviv University, Ramat Aviv 69978, Israel. 3. IVF Unit and Reproduction Lab, Department of Obstetrics and Gynecology, Chaim Sheba Medical Center, Ramat-Gan 52621, Israel. 4. Department of Gynecologic Oncology, Chaim Sheba Medical Center, Ramat Gan 52621, Israel. 5. Institutional Tumor Banks, Chaim Sheba Medical Center, Ramat Gan 52621, Israel. 6. Sackler Faculty of Medicine, Tel-Aviv University, Ramat Aviv 69978, Israel; IVF Unit and Reproduction Lab, Department of Obstetrics and Gynecology, Chaim Sheba Medical Center, Ramat-Gan 52621, Israel. 7. Sheba Cancer Research Center, Chaim Sheba Medical Center, Ramat Gan 52621, Israel; The Dr. Pinchas Borenstein Talpiot Medical Leadership Program 2012, Institute of Oncology, Chaim Sheba Medical Center, Ramat Gan 52621, Israel. Electronic address: Keren.Levanon@sheba.health.gov.il.
Abstract
OBJECTIVES: Ovulation-related inflammation is suspected to have a causal role in ovarian carcinogenesis, but there are no human models to study the molecular pathways. Our aim is to develop such an ex-vivo model based on human fallopian tube (FT) epithelium exposed to human follicular fluid (FF). METHODS: FT epithelium was dissociated from normal surgical specimens. FF was obtained from donors undergoing in-vitro fertilization. The cells were cultured on collagen-coated Transwells and incubated with FF for various periods of time. The transcriptomic changes resulting from FF treatment were profiled using Affymetrix expression arrays. Specific characteristics of the FT pre-cancerous lesions were studied using immunohistochemistry, immunofluorescence, RT-PCR and XTT assay. RESULTS: We show that FF exposure causes up-regulation of inflammatory and DNA repair pathways. Double stranded DNA breaks are induced. There is a minor increase in cell proliferation. TP53, which is the hallmark of the precursor lesion in-vivo, is accumulated. Levels of expression and secretion of Interleukin-8 are significantly increased. CONCLUSIONS: Our model addresses the main non-genetic risk factor for ovarian cancer, namely the impact of ovulation. This study demonstrates the biological implications of in-vitro exposure of human FT epithelial cells to FF. The model replicates elements characterizing the precursor lesions of ovarian cancer, and warrants further investigation of the linkage between repeated exposure to ovulation-related damage and accumulation of neoplastic changes.
OBJECTIVES: Ovulation-related inflammation is suspected to have a causal role in ovarian carcinogenesis, but there are no human models to study the molecular pathways. Our aim is to develop such an ex-vivo model based on human fallopian tube (FT) epithelium exposed to human follicular fluid (FF). METHODS: FT epithelium was dissociated from normal surgical specimens. FF was obtained from donors undergoing in-vitro fertilization. The cells were cultured on collagen-coated Transwells and incubated with FF for various periods of time. The transcriptomic changes resulting from FF treatment were profiled using Affymetrix expression arrays. Specific characteristics of the FT pre-cancerous lesions were studied using immunohistochemistry, immunofluorescence, RT-PCR and XTT assay. RESULTS: We show that FF exposure causes up-regulation of inflammatory and DNA repair pathways. Double stranded DNA breaks are induced. There is a minor increase in cell proliferation. TP53, which is the hallmark of the precursor lesion in-vivo, is accumulated. Levels of expression and secretion of Interleukin-8 are significantly increased. CONCLUSIONS: Our model addresses the main non-genetic risk factor for ovarian cancer, namely the impact of ovulation. This study demonstrates the biological implications of in-vitro exposure of human FT epithelial cells to FF. The model replicates elements characterizing the precursor lesions of ovarian cancer, and warrants further investigation of the linkage between repeated exposure to ovulation-related damage and accumulation of neoplastic changes.
Authors: Mark E Sherman; Ronny I Drapkin; Neil S Horowitz; Christopher P Crum; Sue Friedman; Janice S Kwon; Douglas A Levine; Ie-Ming Shih; Donna Shoupe; Elizabeth M Swisher; Joan Walker; Britton Trabert; Mark H Greene; Goli Samimi; Sarah M Temkin; Lori M Minasian Journal: Cancer Prev Res (Phila) Date: 2016-05-24
Authors: Anthony N Karnezis; Kathleen R Cho; C Blake Gilks; Celeste Leigh Pearce; David G Huntsman Journal: Nat Rev Cancer Date: 2016-11-25 Impact factor: 60.716