沉默Smad4基因对乳腺癌MCF-7细胞增殖和凋亡的影响

doi:10.13481/j.1671-587x.20170506

吉林大学学报(医学版) ›› 2017, Vol. 43 ›› Issue (05): 887-892.doi: 10.13481/j.1671-587x.20170506

沉默Smad4基因对乳腺癌MCF-7细胞增殖和凋亡的影响

刘楠楠¹, 李玉林², 李荣贵², 孙立伟³, 刘学娟⁴

1. 北华大学基础医学院病理学教研室, 吉林 132013;
2. 吉林大学基础医学院病理生物学教育部重点实验室, 吉林长春 130021;
3. 北华大学化学与生物学院生命科学研究中心, 吉林吉林 132013;
4. 吉林大学第一医院病理科, 吉林长春 130021

收稿日期:2017-01-25 出版日期:2017-09-28 发布日期:2017-09-29
通讯作者: 刘楠楠,副教授(Tel:0432-62031704,E-mail:liunn1976@163.com) E-mail:liunn1976@163.com
作者简介:刘楠楠(1976-),女,吉林省吉林市人,副教授,医学博士,主要从事肿瘤分子病理学方面的研究。
基金资助:
国家科技部支撑计划项目资助课题（2012BAI19B05）

Influence of silencing Smad4 gene in proliferation and apoptosis of breast carcinoma MCF-7 cells

LIU Nannan¹, LI Yulin², LI Ronggui², SUN Liwei³, LIU Xuejuan⁴

1. Department of Pathology, School of Basic Medical Sciences, Beihua University, Jilin 132013, China;
2. Key Laboratory of Pathobiology, Ministry of Education, School of Basic Medical Sciences, Jilin University, Changchun 130021, China;
3. Life Science Research Center, School of Chemistry and Biology, Beihua University, Jilin 132013, China;
4. Department of Pathology, First Hospital, Jilin University, Changchun 130021, China

Received:2017-01-25 Online:2017-09-28 Published:2017-09-29

摘要/Abstract

摘要： 目的：研究降低Smad4表达对乳腺癌MCF-7细胞增殖和凋亡的影响，并探讨其可能的作用机制。方法：体外培养人乳腺癌MCF-7和MDA-MB-231细胞，采用逆转录-聚合酶链反应（RT-PCR）法检测MCF-7细胞和MDA-MB-231细胞中Smad4 mRNA表达水平；Smad4-shRNA与Scramble-shRNA质粒分别稳定转染Smad4高表达MCF-7细胞，实验分为未转染MCF-7细胞组（正常对照组），采用Smad4基因沉默组和Scramble组（阴性对照组）；采用CCK-8法检测各组细胞增殖能力，流式细胞术检测各组细胞凋亡率，实时荧光定量PCR（Real-time PCR）法检测增殖相关基因CDKN1A、CDK1和CDK2以及凋亡相关基因Suvivin、bcl-2、caspase3和caspase9 mRNA表达水平。结果：各组细胞增殖能力比较差异无统计学意义（P > 0.05），各组细胞CDKN1A、CDK1和CDK2 mRNA表达水平比较差异无统计学意义（P > 0.05）；与正常对照组和阴性对照组比较，Smad4基因沉默组细胞凋亡率明显降低（P < 0.01），Smad4基因沉默组细胞Suvivin和bcl-2 mRNA表达水平明显升高（P < 0.01），caspase3和caspase9 mRNA表达水平明显降低（P < 0.05）。结论：Smad4可以通过下调Suvivin和bcl-2的表达、上调caspase3和caspase9表达引起细胞凋亡。

关键词: Smad4, 细胞增殖, 细胞凋亡, MCF-7细胞

Abstract: Objective: To study the effects of down-regulated Smad4 expression on the proliferation and apoptosis of breast carcinoma MCF-7 cells, and to explore their mechanisms. Methods: The human breast carcinoma MCF-7 cells and MDA-MB-231cells were cultured in vitro. RT-PCR method was used to detect the expression levels of Smad4 mRNA in MCF-7 cells and MDA-MB-231 cells. The Smad4-shRNA plasmid and Scramble-shRNA plasmid were respectively stably transfected into the MCF-7 cells with high expression of Smad4. The experiment was divided into non-transfected MCF-7 cells (normal control) group,Smad4 gene silencing group, Scramble (negative control)group.The proliferation abilities of the cells in various groups were detected by CCK-8 method. The apoptotic rates of the cells in various groups were detected by flow cytometry. Real-time PCR method was used to detect the mRNA expression levels of the proliferation-related genes CDKN1A, CDK1 and CDK2 and the apoptosis-related genes Suvivin, bcl-2, caspase 3 and caspase 9. Results: The proliferation abilities of cells had no statistical significance between various groups(P>0.05). The mRNA expression levels of CDKN1A, CDK1 and CDK2 in the cells had no statistical significance between various groups (P>0.05).Compared with normal control group and negative control group, the apoptotic rate of the cells in Smad4 gene silencing group was significantly decreased(P<0.01), the expression levels of Suvivin and bcl-2 mRNA in Smad4 gene silencing group were significantly increased(P<0.01), and the mRNA expression levels of caspase 3 and caspase 9 in Smad4 gene silencing group were significantly decreased(P<0.05). Conclusion: Smad4 could induce the apoptosis of MCF-7 cells by down-regulating the expressions of Suvivin and bcl-2 and up-regulating the expressions of caspase 3 and caspase 9.

Key words: Smad4, MCF-7 cells, cell proliferation, apoptosis

中图分类号:

R361.3

刘楠楠, 李玉林, 李荣贵, 孙立伟, 刘学娟. 沉默Smad4基因对乳腺癌MCF-7细胞增殖和凋亡的影响[J]. 吉林大学学报(医学版), 2017, 43(05): 887-892.

LIU Nannan, LI Yulin, LI Ronggui, SUN Liwei, LIU Xuejuan. Influence of silencing Smad4 gene in proliferation and apoptosis of breast carcinoma MCF-7 cells[J]. Journal of Jilin University Medicine Edition, 2017, 43(05): 887-892.

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沉默Smad4基因对乳腺癌MCF-7细胞增殖和凋亡的影响

Influence of silencing Smad4 gene in proliferation and apoptosis of breast carcinoma MCF-7 cells

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