缺氧诱导卵巢上皮性癌细胞形成拟态血管的前期研究  

Primary study of vasculogenic mimicry induced by hypoxia in epithelial ovarian carcinoma

作　　者：尧良清[1] 丰有吉[1] 丁景新[1] 徐丛剑[1] 金鸿雁[1] 殷莲华[2] 

机构地区：[1]复旦大学附属妇产科医院妇科,上海200011 [2]上海医学院生理与病理生理学系

出　　处：《中华妇产科杂志》2005年第10期662-665,共4页Chinese Journal of Obstetrics and Gynecology

基　　金：国家自然科学基金资助项目(30471806);中国博士后科学基金资助项目(20040350454);上海市科学技术委员会科研计划资助项目(04JC14021);上海市医学重点学科建设项目(05Ⅲ016)

摘　　要：目的探讨缺氧在卵巢上皮性癌细胞系SKOV3和ES2细胞拟态血管形成过程中的作用及西罗莫司对其的抑制效应。方法建立人工基底膜基质凝胶Matrigel三维培养系统,分别在常氧(常氧组)、缺氧(缺氧组)、缺氧时加入西罗莫司(缺氧+西罗莫司组)等条件下诱导,观察SKOV3和ES2细胞形成拟态血管的能力;应用相差显微镜和扫描电镜观察拟态血管的形态及其立体结构的特点;用RT-PCR技术检测不同条件下缺氧诱导因子(HIF)1αmRNA的相对表达量。结果培养7d后,缺氧组SKOV3和ES2细胞出现变形、伸展,形成腔样、管状、分支和网络状等管道结构;而常氧组及缺氧+西罗莫司组,部分细胞出现伸展但不形成管道结构。缺氧组SKOV3和ES2细胞的HIF-1αmRNA表达量分别为0·801±0·034和0·736±0·059,显著高于缺氧+西罗莫司组(分别为0·025±0·007和0·231±0·035;P<0·01,P<0·05)和常氧组(分别为0·010±0·004和0·011±0·002;P均<0·01)。结论缺氧是卵巢上皮性癌细胞形成拟态血管的重要诱导因素,西罗莫司通过抑制HIF-1αmRNA的表达阻断拟态血管的形成。Objective To explore possibility of vasculogenic mimicry induced by hypoxia and the inhibitive effect by sirolimus in epithelial ovarian carcinoma in vitro. Methods Based on three-dimensional cell culture system developed by Matrigel, ovarian cell lines SKOV3 and ES2 were induced under conditions of hypoxia, hypoxia added with sirolimus and no-hypoxia, respectively. Potential formation of tumor channels and their characterization of network were observed by light microscopy and scanning electronic microscopy. Relative hypoxia-inducible factor （HIF） lot mRNA expression was detected by RT-PCR simultaneously. Results The micrograph showed both SKOV3 and ES2 cells appeared expanded and re-shaped, then formed blood vessel-like structures such as cavity, channel, branch and network. These capabilities of cells were inhibited by sirolimus and no-hypoxia. The levels of HIF-1 α mRNA expression of SKOV3 and ES2 were 0. 801 ± 0. 034 and 0. 736 ± 0. 059 under hypoxia, which were significantly higher than under hypoxia added with sirolimus （0. 025 ± 0. 007,0. 231 ± 0. 035;P 〈 0. 01, P 〈 0. 05）, and those under no-hypoxia （0. 010 ± 0. 004,0. 011 ± 0. 002; both P 〈 0. 01 ）. Conclusions Hypoxia plays a key role in development of vasculogenic mimicry in epithelial ovarian carcinoma. Sirolimus can inhibit vasculogenic mimicry effectively by blocking HIF-1 α at transcription level.

关 键 词：卵巢肿瘤 新生血管化 病理性 细胞缺氧 西罗莫司 缺氧诱导因子 卵巢上皮性癌 细胞形成 血管形成 MRNA表达量 SKOV3 

分 类 号：R737.31[医药卫生—肿瘤]