上海交通大学学报(医学版)

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CacyBP/SIP基因沉默对人乳腺癌MDA-MB-231细胞凋亡的影响

王 燕1,廉 斌2,吕 叶1,王宁菊1,李少林3   

  1. 1.宁夏医科大学总医院肿瘤内科, 银川 750001; 2.宁夏医科大学总医院肿瘤外科, 银川 750001; 3.重庆医科大学放射医学教研室, 重庆 400016
  • 出版日期:2014-11-28 发布日期:2014-12-02
  • 通讯作者: 王宁菊, 电子信箱: doctorning@163.com; 李少林, 电子信箱: lishaolin@cqmu.edu.cn。
  • 作者简介:王 燕(1974—), 女, 满族, 副主任医师, 博士生; 电子信箱: mgwy1974@163.com。
  • 基金资助:

    国家自然科学基金(81460400, 81171365)

Effects of silencing CacyBP/SIP gene on apoptosis of human breast cancer cell line MDA-MB-231

WANG Yan1, LIAN Bin2, LÜ Ye1, WANG Ning-ju1, LI Shao-lin3   

  1. 1.Department of Oncology, General Hospital, Ningxia Medical University, Yinchuan 750001, China; 2.Department of Surgical Oncology, General Hospital, Ningxia Medical University, Yinchuan 750001, China; 3.Department of Radiological Medicine, Chongqing Medical University, Chongqing 400016, China
  • Online:2014-11-28 Published:2014-12-02
  • Supported by:

    National Natural Science Foundation of China, 81460400, 81171365

摘要:

目的 观察干扰小RNA(siRNA)介导的CacyBP/SIP基因沉默对人乳腺癌MDA-MB-231细胞凋亡的影响。方法 化学合成CacyBP/SIP基因序列特异性siRNA(CacyBP/SIP siRNA),采用脂质体法转染siRNA至MDA-MB-231细胞(CacyBP/SIP siRNA组),MTT比色法检测细胞增殖,流式细胞仪检测细胞凋亡率。Real-time PCR和Western blotting检测MDA-MB-231细胞内Bcl-2、Bax、Caspase-3 mRNA及蛋白的表达。另设阴性对照组和空白对照组。结果 与阴性对照组和空白对照组比较,CacyBP/SIP siRNA组MDA-MB-231细胞增殖能力显著下降(P<0.05),细胞凋亡率显著增高(P<0.05)。Caspase-3和Bax mRNA及蛋白表达上调(P<0.05),Bcl-2 mRNA及蛋白表达下调(P<0.05)。结论 靶向沉默乳腺癌MDA-MB-231细胞中CacyBP/SIP基因表达后,可以抑制乳腺癌细胞增殖并诱导凋亡,其作用可能通过下调Bcl-2表达,上调Caspase-3和Bax表达发挥诱导细胞凋亡的作用。

关键词: 乳腺癌, CacyBP/SIP基因, 干扰小RNA, 细胞凋亡

Abstract:

Objective To observe the effects of small interfering RNA (siRNA)-mediated silencing CacyBP/SIP gene on the apoptosis of human breast cancer cell line MDA-MB-231. Methods Chemically synthesized siRNAs targeting CacyBP/SIP (CacyBP/SIP siRNA) was transfected to MDA-MB-231 cells by LipofectAMINTM2000 (CacyBP/SIP siRNA group). The proliferation of MDA-MB-231 cells was detected by the MTT assay. The apoptosis rate was detected by the flow cytometry. The mRNA and protein expressions of Bcl-2, Bax, and Capase-3 were detected by the Realtime PCR and Western blotting respectively. Besides, the negative control group and blank control group were also established. Results Compared to the negative control group and blank control group, the proliferation rate of MDA-MB-231 cells of the CacyBP/SIP siRNA group significantly decreased (P<0.05), and the apoptosis rate significantly increased (P<0.05). The mRNA and protein expressions of Caspase-3 and Bax significantly increased (P<0.05), while the mRNA and protein expressions of Bcl-2 significantly decreased (P<0.05). Conclusion Targeted silencing CacyBP/SIP gene of breast cancer cell line MDA-MB-231 inhibits the proliferation of MDA-MB-231 cells and induces apoptosis. The mechanism of inducing apoptosis may be relevant to the down-regulation of Bcl-2 expression and up-regulation of Caspase-3 and Bax.

Key words: breast cancer, CacyBP/SIP gene, small interfering RNA, cell apoptosis