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作 者:冯旺琴[1] 崔竹梅[1] 冯凤芝[2] 祈秀娟[1] 丁芳[3] 李文东[3] 刘芝华[3]
机构地区:[1]青岛大学医学院附属医院妇产科 [2]中国医学科学院中国协和医科大学北京协和医院妇产科 [3]中国医学科学院肿瘤研究所分子肿瘤学国家重点实验室
出 处:《中华妇产科杂志》2005年第8期553-557,共5页Chinese Journal of Obstetrics and Gynecology
摘 要:目的探讨SLP-2mRNA在子宫内膜癌中的表达,及其在子宫内膜癌发生、发展中的作用。方法采用半定量RT-PCR技术检测32例子宫内膜癌组织和28例正常子宫内膜组织中SLP-2mRNA的表达情况;并用表达SLP-2基因的正义和反义质粒,分别转染子宫内膜癌细胞系HEC-1B细胞,RT-PCR技术检测转染前、后HEC-1B细胞中SLP-2mRNA的表达情况,观察转染前、后细胞的形态变化;用四甲基偶氮唑蓝(MTT)法检测细胞生长情况,并绘制细胞生长曲线;流式细胞仪检测细胞周期的变化。结果子宫内膜癌组织中SLP-2mRNA的表达水平为1.6±0.7,正常子宫内膜组织为0.7±0.3,两者比较,差异有统计学意义(P<0.05)。表达SLP-2基因的正义和反义质粒成功转入HEC-1B细胞中,正义质粒转染后SLP-2mRNA表达水平增加约2.4倍,细胞生长变快,其中G1期减少12.5%,S期增高8.0%;反义质粒转染后SLP-2mRNA表达水平减少50%,细胞生长变慢,其中G1期增高10.5%,S期减少9.8%。结论子宫内膜癌中SLP-2mRNA的高表达与子宫内膜癌发生、发展有一定关系。Objective To characterize the differential expression of SLP-2 in endometrial cancer, and to study the effect of human SLP-2 gene on human endometrial cancer cell line. Methods The expression of SLP-2 gene in 32 eases of endometrial cancer and 28 eases of normal endometrial tissues was examined by semi-quantitative RT-PCR. Eukaryotie expression vectors of sense and antisense SLP-2 were constructed and transfected into HEC-1B cell line using lipofeetamine 2000 respectively. The morphological changes of the cell were observed by phase contrast microscopy. The cell growth was detected by methyl thiazolyl tetrazolium (MTT) assay, and the cell cycles were analyzed by flow cytometry. Results The expression of SLP-2 mRNA in endometrial cancer tissues was higher than that in normal endometrial tissues ( 1.6 ± 0. 7 vs 0. 7 ± 0. 3, P 〈 0. 05). Sense and antisense human SLP-2 constructs were transfeeted into HEC-1B cell line respectively. After being transfeeted with sense SLP-2, the expression of SLP-2 mRNA in HEC-1B cell line was increased by about 2. 4 times that of the control group, the cell growth was accelerated, and the number of ceils in G1 phase was decreased by 12. 5% , S phase was increased by 8.0%. After being transfected with antisense SLP-2, the expression of SLP-2 mRNA was declined 50%. The transfected ceils showed slower growth, and the number of cells in G1 phase was significantly increased by 10. 5%, and S phase was declined by 9. 8%. Conclusion SLP-2 mRNA shows up-regulation in endometrial cancer tissues, and it may have some relationship with carcinogenesis of endometrial cancer.
关 键 词:子宫内膜肿瘤 蛋白质类 逆转录聚合酶链反应 转染 mRNA表达 子宫内膜癌 PCR技术检测 细胞生长曲线 其意义 表达及
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