Sheba Cancer Research Center, Chaim Sheba Medical Center, Ramat Gan 52621, Israel.
Sheba Cancer Research Center, Chaim Sheba Medical Center, Ramat Gan 52621, Israel; Sackler Faculty of Medicine, Tel-Aviv University, Ramat Aviv 69978, Israel.
Gynecol Oncol. 2014 Feb;132(2):322-7. doi: 10.1016/j.ygyno.2013.12.015. Epub 2013 Dec 17.
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).
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.
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.
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.
排卵相关炎症被怀疑在卵巢癌发生中起因果作用，但目前尚无人类模型来研究其分子途径。我们的目的是基于暴露于人卵泡液（FF）中的人输卵管（FT）上皮细胞，开发这样一种离体模型。
从正常手术标本中分离 FT 上皮细胞。FF 从接受体外受精的供体中获得。将细胞培养在涂有胶原蛋白的 Transwell 上，并与 FF 孵育不同的时间。使用 Affymetrix 表达谱分析 FF 处理后产生的转录组变化。使用免疫组织化学、免疫荧光、RT-PCR 和 XTT 测定研究 FT 癌前病变的特定特征。
我们表明 FF 暴露会导致炎症和 DNA 修复途径的上调。诱导双链 DNA 断裂。细胞增殖略有增加。TP53 是体内前病变的标志，积累。白细胞介素-8 的表达和分泌水平显著增加。
我们的模型解决了卵巢癌的主要非遗传风险因素，即排卵的影响。本研究证明了人 FT 上皮细胞体外暴露于 FF 的生物学意义。该模型复制了卵巢癌前病变的特征性元素，并进一步证明了反复暴露于排卵相关损伤与肿瘤变化的积累之间的联系。
