S腺苷甲硫氨酸脱羧酶基因沉默抑制乳腺癌MCF7细胞生长的研究

山东大学学报(医学版) ›› 2009, Vol. 47 ›› Issue (12): 66-69.

S腺苷甲硫氨酸脱羧酶基因沉默抑制乳腺癌MCF7细胞生长的研究

辛佳璇¹，陈世敏^1,2，王伟¹，阎云飞¹，徐春晓¹，刘斌¹，刘贤锡¹

1. 山东大学医学院生物化学与分子生物学研究所，济南 250012；
2. 济南军区总医院实验诊断科，济南 250031

收稿日期:2009-04-02 出版日期:2009-12-16 发布日期:2009-12-16
通讯作者: 刘贤锡（1945- ），男，博士生导师，主要从事肿瘤分子生物学的研究。 Email:xianxi@sdu.edu.cn
作者简介:辛佳璇（1985- ），女，硕士研究生，主要从事肿瘤分子生物学的研究。 Email:jiaxuanxin@yahoo.com.cn
基金资助:
山东省科技攻关项目(2008GG30002030)。

Gene silencing of Sadenosylmethionine decarboylase inhibits growth of breast cancer MCF7 cells

XIN Jiaxuan ¹，　CHEN Shimin ^1,2，　WANG Wei ¹，　YAN Yunfei ¹，　XU Chunxiao ¹，　LIU Bin ¹，　LIU Xianxi ¹

1. Institute of Biochemistry and Molecular Biology， School of Medicine， Shandong University， Jinan 250012， China；
2. Department of Clinical Laboratory, General Hospital of Jinan Military Region， Jinan 250031， China

Received:2009-04-02 Online:2009-12-16 Published:2009-12-16

摘要/Abstract

摘要：

目的探讨S腺苷甲硫氨酸脱羧酶（SAMDC）基因沉默对乳腺癌MCF7细胞增殖及侵袭能力的抑制作用。方法用特异性沉默SAMDC基因表达的RNA干扰真核表达载体SAMDC shRNA表达载体(pGPU6SAMDC)转染乳腺癌MCF7细胞，采用RTPCR和Westernblot技术分别检测SAMDC mRNA和蛋白质表达水平；细胞增殖实验（CCK8法）和细胞周期分析评价SAMDC基因沉默对乳腺癌MCF7细胞增殖的影响，肿瘤侵袭实验观察SAMDC基因沉默对乳腺癌细胞MCF7侵袭活性的影响。结果SAMDC shRNA表达载体转染后，MCF7细胞SAMDC mRNA水平下调43.8％，蛋白表达水平下降66.5％；MCF7细胞的增殖活性和侵袭能力均受到明显抑制，细胞增殖活性下降37%，细胞周期分析G1期细胞增加。结论SAMDC基因沉默可通过延长细胞周期的G1期而抑制乳腺癌MCF7细胞的增殖,并能降低癌细胞侵袭活性，提示SAMDC可作为乳腺癌基因治疗的靶分子。

关键词: 乳腺肿瘤；S腺苷甲硫氨酸脱羧酶；RNA干扰；基因治疗

Abstract:

To investigate the inhibitory effect of the Sadenosylmethionine decarboylase (SAMDC) gene silencing on proliferation and invasion of breast cancer MCF7 cells. MethodsBreast cancer MCF7 cells were transfected with a short hairpin RNA eukaryotic expression vector against SAMDC（pGPU6SAMDC）. The effect of suppressing proliferation of MCF7 cells was detected by cell proliferation assay(CCK8) and cell cycle analysis. Semiquantitative RTPCR analysis of mRNA and western blot analysis of proteins were used to determine expression of the SAMDC gene. The invasive ability of breast cancer MCF7 cells was assessed by invasion assay in vitro. ResultsAfter the shRNA expression vector was transfected into breast cancer MCF7 cells, the growth and invasive ability of MCF7cells were significantly inhibited, and the proliferation of cells was decreased by 37%. Gene expression of SAMDC in MCF7 cells was also significantly inhibited, of mRNA was inhibited by 43.8% and of protein was inhibited by 66.5%. Moreover, the proportion of MCF7 cells in the G0/G1 phase was increased. ConclusionThe shRNA of the SAMDC gene can effectively inhibit the proliferation and invasion of MCF7 cells and is a potential target in gene therapy of breast cancer.

Key words: Breast neoplasms; Sadenosylmethionine decarboylase; RNA interference; Gene therapy

中图分类号:

Q784

辛佳璇1，陈世敏1,2，王伟1，阎云飞1，徐春晓1，刘斌1，刘贤锡1. S腺苷甲硫氨酸脱羧酶基因沉默抑制乳腺癌MCF7细胞生长的研究[J]. 山东大学学报(医学版), 2009, 47(12): 66-69.

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