下调CENP-E对乳腺癌MCF-7细胞的影响

中国生物工程杂志

2012, Vol. 32

Issue (10): 7-13

研究报告

下调CENP-E对乳腺癌MCF-7细胞的影响

湛晓琴, 胡敏, 宋培培, 章帆, 刘晨, 施琼, 翁亚光

重庆医科大学检验医学院临床检验诊断学教育部重点实验室重庆 400016

Effect of Low Expression of CENP-E on Breast Cancer Cell Line MCF-7

ZHAN Xiao-qin, HU Min, SONG Pei-pei, ZHANG Fan, LIU Chen, SHI Qiong, WENG Ya-guang

Key Laboratory for Clinical Laboratory Diagnostics of Education Ministry, College of Laboratory Medical, Chongqing Medical University, Chongqing 400016, China

全文: PDF(1378 KB) HTML

摘要： 目的:探讨有丝分裂检查点蛋白着丝粒蛋白-E(CENP-E)基因在肿瘤发生发展中的作用。方法:利用shRNA下调CENP-E基因的表达,分别用巢式PCR和Western blot检测CENP-E mRNA和蛋白的表达;MTT检测CENP-E下调后MCF-7细胞的增殖变化;流式细胞术检测CENP-E下调后对MCF-7细胞凋亡的影响;Transwell试验检测MCF-7细胞的迁移和侵袭能力变化;间接免疫荧光检测细胞内CENP-E蛋白和有丝分裂情况。结果:shRNA能有效抑制CENP-E mRNA和蛋白的表达。MTT结果显示CENP-E下调后MCF-7细胞的增殖能力减弱(P<0.05);流式细胞术显示下调CENP-E后能促进MCF-7细胞的凋亡;间接荧光结果显示CENP-E干扰后MCF-7细胞内CENP-E蛋白减少并伴有核分裂异常;Transwell试验显示CENP-E干扰组细胞的迁移和侵袭能力增强(P<0.05)。结论:下调部分CENP-E的表达能抑制MCF-7细胞的增殖,促进MCF-7细胞的凋亡,增强MCF-7细胞的迁移和侵袭能力。

关键词： 非整倍体; 有丝分裂检查; CENP-E; 肿瘤

Abstract: Objective To explore the role of mitotic protein CENP-E in cancer.Methods Plasmid vector containing CENP-E special short hair RNA(shRNA)was transfected into human breast cancer MCF-7 cells.Nest PCR and Western blot was used to detect CENP-E mRNA and protein level;the proliferation of cells with down-regulated CENP-E were analyzed by MTT assay;the apoptosis of cells were detected by flow cytometry(FCM);the migration and invasion ability of cells were evaluated by transwell assay;indirect immunofluorescence was used to detect CENP-E protein and cell mitosis.Results Compared with the control group,the CENP-E mRNA and protein level in MCF-7 cells transfected shRNA-CENP-E were decreased; the cell growth were inhibited in shRNA-CENP-E group(P<0.05);the results of FCM showed that the cell apoptosis ratio were obviously increased after down-regulated CENP-E;indirect immunofluorescence indicated CENP-E protein was reduced after transfected with shRNA-CENP-E,and the nucleus showed abnormal caryogram;but the cell ability of migration and invasion in shRNA-CENP-E group were increased(P<0.05). Conclusion: Down-regulated part of CENP-E gene expression in breast cancer MCF-7 cells can inhibit cell proliferation,induce cell apoptosis,meanwhile enhance the cell ability of migration and invasion.

Key words: Aneuploidy Mitotic checkpoint CENP-E Tumor

收稿日期: 2012-07-19 出版日期: 2012-10-25

ZTFLH:

Q751

基金资助: 国家自然科学基金(30872770);教育部高等学校博士学科点专项科研基金(20115503110009)资助项目

通讯作者: 翁亚光,电子信箱:yaguangweng@126.com E-mail: yaguangweng@126.com

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引用本文:

湛晓琴, 胡敏, 宋培培, 章帆, 刘晨, 施琼, 翁亚光. 下调CENP-E对乳腺癌MCF-7细胞的影响[J]. 中国生物工程杂志, 2012, 32(10): 7-13.

ZHAN Xiao-qin, HU Min, SONG Pei-pei, ZHANG Fan, LIU Chen, SHI Qiong, WENG Ya-guang. Effect of Low Expression of CENP-E on Breast Cancer Cell Line MCF-7. China Biotechnology, 2012, 32(10): 7-13.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2012/V32/I10/7

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