缺氧诱导上皮性卵巢癌细胞分化逆转机制的实验研究  

Molecular mechanism of retrodifferentiation from epithelial ovarian cancer cells induced by hypoxia

作　　者：陈默[1] 殷啸俊 尧良清[1] 李娜[1] 竺鹏飞[1] 徐丛剑[1] 

机构地区：[1]复旦大学附属妇产科医院妇科,上海200001 [2]昆山市第二人民医院妇科,江苏昆山215300

出　　处：《中国病理生理杂志》2011年第5期848-852,共5页Chinese Journal of Pathophysiology

基　　金：上海市科委自然科学基金资助项目(No.08ZR1401900);高等学校博士学科点专项科研基金资助项目(No.20070246020)

摘　　要：目的:在三维培养系统中进行缺氧刺激,可诱导卵巢恶性上皮性肿瘤细胞SKOV-3和ES-2向血管内皮样细胞分化逆转,本文拟探讨其相关的分子机制。方法:建立Matrigel三维培养系统,在1%O2缺氧条件下,诱导卵巢上皮性癌细胞SKOV-3和ES-2向血管内皮样细胞分化逆转后,应用端粒重复序列扩增法(TRAP)和实时荧光定量RT-PCR检测原始肿瘤细胞、肿瘤内皮样细胞和人脐静脉内皮细胞(HUVECs)端粒酶活性以及周期蛋白cyclinD1、抑癌基因p53、原癌基因v-src转录水平的差异。结果:常氧时,SKOV-3和ES-2细胞端粒酶活性表达为阳性,缺氧诱导后SKOV-3内皮样细胞端粒酶活性为阳性,而ES-2内皮样细胞端粒酶活性转为阴性。缺氧诱导后的SKOV-3和ES-2内皮样细胞cyclinD1 mRNA的表达显著低于常氧时SKOV-3和ES-2的表达(P<0.05或P<0.01);和HUVECs的表达无显著差异。SKOV-3和ES-2内皮样细胞p53 mRNA的表达显著高于常氧时SKOV-3、ES-2和HUVECs的表达(P<0.05或P<0.01)。缺氧和常氧状态下各种细胞均不表达v-src mRNA。结论:缺氧可以诱导少量卵巢上皮性癌细胞SKOV-3和ES-2向血管内皮样细胞分化逆转,主要是通过改变端粒酶的活性,调节cyclinD1和p53的转录水平发挥调控作用。AIM： To investigate the molecular mechanism of retrodifferentiation from epithelial ovarian cancer into vascular endothelial-like cells induced by hypoxia.METHODS： The three-dimensional culture system with Matrigel was established to culture SKOV-3 and ES-2 ovarian cancer cell lines under hypoxic condition（1% O2）.Telomeric repeat amplification protocol（TRAP） and real-time RT-PCR were used to analyze the activity of telomerase and the mRNA expression of cyclin D1,p53 and v-src in endothelial-like cells differentiated from the microdissected tumors.The data were compared with those in original cancer cells and human umbilical vein endothelial cells（HUVECs）.RESULTS： The activity of telomerase was positive in SKOV-3 and ES-2 cells under normoxic condition,and was positive in SKOV-3 endothelial-like cells but negative in ES-2 endothelial-like cells under hypoxic condition.The mRNA expression of cyclin D1 in SKOV-3 and ES-2 endothelial-like cells with hypoxia was significantly lower than that in the same cells with normoxia（P0.01）,but was not significantly different from that in HUVECs（P0.05）.In SKOV-3 and ES-2 endothelial-like cells,the mRNA expression of p53 was higher with hypoxia than that with normoxia.No expression of v-src mRNA in all kinds of the cells under either hypoxic or normoxic condition was detected.CONCLUSION： Hypoxia induces the differentiation of SKOV-3 and ES-2 cells into vascular endothelial-like cells by decreasing telomerase activity and cyclin D1 expression,and the increasing the mRNA expression of p53.

关 键 词：卵巢肿瘤 缺氧 血管内皮样细胞 细胞分化 端粒酶 细胞周期蛋白D1 基因 p53 基因 v-src 

分 类 号：R737.31[医药卫生—肿瘤]