[1]杨德辉,陈雪梅,何俊琳,等.hsa-miR-200a对HEC-1B人子宫内膜腺癌细胞系增殖与凋亡的影响[J].第三军医大学学报,2012,34(16):1608-1612.
 Yang Dehui,Chen Xuemei,He Junlin,et al.Effect of hsa-miR-200a on proliferation and apoptosis in human endometrium adenocarcinoma cell line HEC-1B[J].J Third Mil Med Univ,2012,34(16):1608-1612.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
34卷
期数:
2012年第16期
页码:
1608-1612
栏目:
论著
出版日期:
2012-08-30

文章信息/Info

Title:
Effect of hsa-miR-200a on proliferation and apoptosis in human endometrium adenocarcinoma cell line HEC-1B
作者:
杨德辉陈雪梅何俊琳王应雄丁裕斌李荣刘学庆
重庆医科大学公共卫生学院生殖生物学研究室
Author(s):
Yang Dehui Chen Xuemei He Junlin Wang Yingxiong Ding Yubin Li Rong Liu Xueqing
Laboratory of Reproductive Biology, School of Public Health, Chongqing Medical University, Chongqing, 400016, China
关键词:
子宫内膜癌hsa-miR-200aPTEN增殖与凋亡
Keywords:
endometrial carcinoma hsa-miR-200a PTEN cell proliferation apoptosis
分类号:
R394.2;R730.23;R737.33
文献标志码:
A
摘要:
目的      探讨hsa-miR-200a对人子宫内膜腺癌HEC-1B细胞增殖与凋亡的影响及其作用机制。      方法      应用荧光实时定量PCR(FQ-PCR)法检测hsa-miR-200a在人子宫内膜腺癌细胞HEC-1B中的表达水平;人工合成hsa-miR-200a模拟物、抑制剂,分别转染HEC-1B细胞;采用MTT法、流式细胞仪检测上调及抑制hsa-miR-200a的表达对HEC-1B细胞增殖活性与凋亡的影响;运用生物信息学分析hsa-miR-200a指向的与增殖和凋亡相关的靶基因;Western blot、FQ-PCR检测转染前后靶蛋白及其mRNA的表达水平。      结果      子宫内膜腺癌细胞HEC-1B表达hsa-miR-200a;FQ-PCR检测hsa-miR-200a模拟物、抑制剂转染HEC-1B细胞成功;MTT法检测结果表明hsa-miR-200a抑制剂转染组中细胞增殖抑制明显;流式细胞仪检测结果表明hsa-miR-200a抑制剂转染组中细胞凋亡增加;生物信息学分析结果表明hsa-miR-200a调控的靶基因是SON、Pdcd4、磷酸酶-张力蛋白同源基因(phosphatase and tensin homolog deleted on chromosome ten,PTEN);靶蛋白PTEN在转染模拟物组表达下调,抑制剂组中表达上调,且转染前后靶基因mRNA水平无显著变化,而蛋白SON、Pdcd4的表达量没有显著变化。      结论      HEC-1B细胞中hsa-miR-200a表达与PTEN蛋白表达呈负相关,表明hsa-miR-200a可能通过调节PTEN蛋白表达水平从而影响HEC-1B细胞的增殖与凋亡。
Abstract:
Objective      To investigate the effect of hsa-miR-200a on the proliferation and apoptosis in human endometrium adenocarcinoma cell line HEC-1B and the potential mechanisms.       Methods      The expression of hsa-miR-200a in HEC-1B cell line was measured by real-time fluorescence quantitative PCR (FQ-PCR). The hsa-miR-200a mimics and inhibitor were synthesized and transferred into HEC-1B cell line by lipofectamine, respectively, and the cell proliferation and apoptosis were detected by methyl-thiazolyl-tetrazolium (MTT) assay and flow cytometry (FCM). Bioinformatics analysis was used to predict the target genes of has-miR-200a on cell proliferation and apoptosis. Western blotting and FQ-PCR were used to determine the protein and mRNA expression of phosphatase and tensin homolog deleted on chromosome ten (PTEN).       Results      Hsa-miR-200a expression was observed in HEC-1B cells. The hsa-miR-200a mimics and inhibitor were successfully transferred into HEC-1B cells, and the expression of hsa-miR-200a was significantly changed after the transfection. Compared with those of the blank control group and negative control group,HEC-1B cell proliferation was significantly inhibited in the miR-200a inhibitor group. The FCM results showed that the apoptotic rate increased in the hsa-miR-200a inhibitor group. The bioinformatics analysis results showed that SON, Pdcd4 and PTEN were the target genes of has-miR-200a. PTEN protein expression significantly decreased in the HEC-1B cells transfected with hsa-miR-200a mimics, and significantly increased in the HEC-1B cells transfected with hsa-miR-200a inhibitor. SON and Pdcd4 protein expression did not change before and after transfection.       Conclusion      Hsa-miR-200a is negatively correlated with PTEN protein expression in HEC-1B cell line, indicating that hsa-miR-200a may affect the proliferation and apoptosis in HEC-1B cells by regulating PTEN protein expression.

参考文献/References:

杨德辉, 陈雪梅, 何俊琳, 等. hsa-miR-200a对HEC-1B人子宫内膜腺癌细胞系增殖与凋亡的影响[J].第三军医大学学报,2012,34(16):1608-1612.

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更新日期/Last Update: 2012-07-29