Nanog基因在卵巢癌和卵巢肿瘤干细胞中的表达及意义  

Expression and Significance of Nanog Gene in Ovarian Cancer and Ovarian Cancer Stem Cells

作　　者：赵乐[1] 纪新强[1] 刘静[1] 武加利[2] 张文卿[3] 

机构地区：[1]青岛大学医学院附属医院妇科,山东青岛266003 [2]山东省济宁市第一人民医院妇产科,山东济宁272000 [3]青岛大学医学院病原生物学教研室,山东青岛266003

出　　处：《现代生物医学进展》2012年第14期2642-2646,共5页Progress in Modern Biomedicine

基　　金：山东省自然科学基金(Y2008c33)

摘　　要：目的:探讨胚胎干细胞关键因子Nanog基因mRNA及其蛋白在卵巢癌和卵巢癌肿瘤干细胞中的表达及意义。方法:选取10例正常卵巢上皮组织、10例卵巢良性肿瘤及60例卵巢癌组织,采用逆转录酶-聚合酶链反应(RT-PCR)方法和免疫组织化学PV-6000两步法检测Nanog mRNA和蛋白表达水平;采用无血清悬浮培养法从SKOV-3卵巢癌细胞株中分离培养肿瘤干细胞,流式细胞术鉴定肿瘤干细胞CD117表达,采用RT-PCR和Western Blot方法检测SKOV-3卵巢癌细胞及肿瘤干细胞中NanogmRNA及其蛋白的表达水平。结果:Nanog mRNA在卵巢癌组织中的表达水平均高于正常卵巢组织和卵巢良性肿瘤组织(P<0.05);Nanog mRNA在不同分化程度及临床分期的卵巢癌组织中表达水平不同,低分化组高于高分化组(P<0.05);III-IV期高于I-II期(P<0.05);免疫组化结果同RT-PCR。从SKOV-3卵巢癌细胞株中成功分离出肿瘤干细胞,SKOV-3卵巢癌细胞和肿瘤干细胞Nanog mRNA相对含量分别为0.6044±0.0368,0.8736±0.0537,差异具有统计学意义(P<0.05),两种细胞Nanog蛋白相对含量分别为0.6364±0.0169 1.2788±0.0314,差别具有统计学意义(P<0.05)。结论:Nanog基因在卵巢癌组织和SKOV-3细胞系中均高表达,其在组织中的表达强度与临床分期及病理分级关系密切,且在肿瘤干细胞中表达高于一般卵巢癌细胞,其与卵巢癌的发生发展关系密切,可能是卵巢癌干细胞的表面标志物,有望成为新的标志物。Objective： To explore the expression of embryonic stem cells key transcription factor Nanog mRNA and protein in ovarian cancer and cancer stem cells.Methods： Reverse transcription polymerase chain reaction（RT-PCR） and immunohistochemistry were performed to detect the expression of Nanog gene in 10 cases of normal ovaries,10 cases of benign ovarian tumors and 60 cases of ovarian cancer.Ovarian caner stem cells were isolated from ovarian cell line SKOV-3 and cultured in serum free medium by nanosphere suspension.Flow cytometry analysis was used to examine the expression of the cell surface marker CD117 in SKOV-3 and ovarian cancer stem cells.We examined the expression of Nanog gene in SKOV-3 and ovarian cancer stem cells by RT-PCR and Western Blot.Results： Nanog gene expression in ovarian cancer was significantly higher than in normal ovarian tissue and benign ovarian tumors（P〈0.05）.The Nanog expression was higher in ovarian cancer with poorly differentiated grade or advanced stage than well differentiated or early stage,respectively（P〈0.05）.Ovarian caner stem cells were isolated,the relative level of Nanog mRNA was 0.6044±0.0368 in SKOV-3 and 0.8736±0.0537 in ovarian tumor stem cell（P〈0.05）.The relative level of Nanog protein was 0.6364±0.0169 in SKOV-3,and 1.2788±0.0314 in ovarian tumor stem cell（P〈0.05）.Conclusions： Nanog gene is highly expressed in ovarian cancer and SKOV-3,which is associated with the differentiation and clinic stage.And Nanog gene is more highly expressed in ovarian cancer stem cell than SKOV-3,which is associated with the development and progression of ovarian cancer.Nanog maybe the antigen marker of ovarian cancer stem cells and could be as novel marker for ovarian cancer.

关 键 词：卵巢癌 NANOG 肿瘤干细胞 基因表达 悬浮培养 

分 类 号：R737.31[医药卫生—肿瘤]