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作 者:高蜀君[1] 丰有吉[1] 殷莲华[2] 孙红[1] 隋龙[1]
机构地区:[1]复旦大学附属妇产科医院妇科,上海200011 [2]复旦大学上海医学院生理和病理生理学系
出 处:《现代妇产科进展》2007年第10期746-749,共4页Progress in Obstetrics and Gynecology
基 金:国家自然科学基金(No:30471806);上海市医学重点学科建设项目-妇产科学(No:05-Ⅲ0169)资助
摘 要:目的:探讨缺氧/复氧后卵巢浆液性囊腺癌细胞株SKOV3增殖能力的改变及其相关机制。方法:SKOV3细胞分别缺氧24h及缺氧后复氧培养不同时间。用MTT法检测细胞的增殖,流式细胞仪检测细胞周期;Westernblot和RT·PCR检测缺氧诱导因子-1α(HIF-1α)的表达。用HIF-1α抑制剂雷帕霉素(Rapamycin)预处理细胞30min后,检测上述各指标的变化。结果:复氧24h细胞增殖率增加(1.38±0.60,P=0.023);复氧48h细胞增殖率恢复为常氧培养水平(0.67±0.28,P=0.48)。流式细胞仪分析显示复氧24h时S期细胞增加[(48.45±1.15)%,P=0.013],G1 期细胞减少[(45.71±2.28)%,P=0.044]。常规培养的SKOV3稳定表达HIF-1α,缺氧可使HIF—1α蛋白表达增加(P=0.0045),复氧8h,HIF—1α表达迅速下降。Rapamycin可以使细胞周期停滞于Go—G1期,从而降低细胞的增殖能力。结论:缺氧/复氧可增强卵巢癌细胞株SKOV3的增殖能力,HIF-1α通路可能在其中起重要作用。Objective:To investigate the influence of anoxia/reoxygenation on the proliferation ability of ovarian cancer cell line SKOV3. Methods: Ovarian cancer cell line SKOV3 was cultured with or without rapamycin in anoxia for 24 hours, reoxygenation 24 and 48 hours after 24 hours hypoxia respectively. MTT was used to detect the ability of cell proliferation. Ceil cycle distribution was examined by flow cytometry (FCM). Expression of HIF-lα was checked by Western blot and reverse transcription-polymerase chain reaction (RT-PCR). Results: The proliferation ability of SKOV3 increased significantly when cultured by reoxygenation for 24hour ( 1.38 ± 0.60 ,P = 0.023 ). FCM analyses showed that SKOV3 cell cultured by 24hour reoxygenation had an increased S phase [ (48.45 ± 1.15)% ,P =0. 013] and decreased Glphase [ ( 45.71 ± 2.28 ) %, P = 0.044 ]. Rapamycin had a G1 phase arrest on SKOV3 cell. Expression of HIF-lα protein increased when cells were exposed to anoxia and reached peak when anoxia for 16h( P = 0.0045 ). It decreased rapidly when reoxygenation 8hour but resumed when reoxy- genation time prolonged gradually. Conclusion :Anoxia/Reoxygenation could increase the proliferation ability of SKOV3. HIF-1α pathway may play an important role in ovarian cancer progression.
分 类 号:R373.31[医药卫生—病原生物学]
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