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中国癌症防治杂志

中国癌症防治杂志 ›› 2012, Vol. 4 ›› Issue (4): 319-323.doi: 10.3969/j.issn.1674-5671.2012.04.05

• 基础研究 • 上一篇    下一篇

干扰赖氨酰氧化酶基因对乳腺癌MDA-MB-231细胞生长、增殖的影响

  

  1.  广西医科大学附属肿瘤医院乳腺外科；广西医科大学研究生学院
  • 出版日期:2012-12-25 发布日期:2013-01-11
  • 通讯作者: 刘剑仑 E-mail:jianlunliu@hotmail.com
  • 基金资助:

     广西科学研究与技术开发计划资助课题（桂科攻0816004-14）

Gene silencing of lysyl oxidase（LOX） by RNA interference：effects on cellular prolifera-tion in breast cancer cell line MDA-MB-231

  • Online:2012-12-25 Published:2013-01-11

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摘要： 目的 探讨抑制赖氨酰氧化酶（lysyl oxidase，LOX）基因表达对人乳腺癌MDA-MB-231细胞生长、增殖的影响及其机制。方法 将乳腺癌MDA-MB-231细胞分为实验组（RNAi组）、阴性对照组（Mock组）和空白对照组（Con组）；设计合成特异性慢病毒干扰载体（LOX-RNAi-LV）转染MDA-MB-231细胞。以荧光定量PCR检测LOX mRNA及增殖因子Ki-67、cyclin D1的表达；Western blot法检测LOX基因蛋白的表达水平；四甲基偶氮唑蓝（MTT）法、细胞集落形成实验观察各组细胞生长及增殖情况。结果 转染LOX-RNAi-LV后，MDA-MB-231细胞中LOX mRNA和蛋白表达水平明显降低，细胞生长、增殖能力明显减弱，细胞集落形成率明显降低，相关增殖因子Ki-67和cyclin D1 mRNA的表达均明显下降（P＜0.05）。结论 转染LOX-RNAi-LV可有效干扰乳腺癌MDA-MB-231细胞中LOX mRNA和蛋白的表达，抑制肿瘤细胞的生长、增殖，其作用机制可能与下调Ki-67、cyclin D1的表达有关。

关键词: 乳腺肿瘤, 赖氨酰氧化酶, RNA干扰, 细胞增殖, 增殖因子

Abstract: Objective To explore the effects of lysyl oxidase（LOX） silencing on proliferation in the breast cancer cell line MDA-MB-231. Methods MDA-MB-231 cultures were divided into three test groups:the RNAi group，the Mock negative control group，and the Con control group，LOX， Ki-67 and cyclin D1 mRNA expression was analyzed by real-time RT-PCR，and LOX protein expression was analyzed by Western blot. Cell growth and proliferation of transfected cells were measured by MTT and cell colony formation assays. Results Expression of both LOX mRNA and protein was significantly down-regulated in the RNAi group. This group also showed significantly less cell growth，less cell colony formation，and lower levels of Ki-67 and cyclin D1 mRNA than the Mock and Con groups（P<0.05）. Conclusions Transfecting MDA-MB-231 cells with LOX-RNAi-LV can significantly interfere with LOX mRNA and protein expression，inhibiting growth and proliferation of breast cancer cells.These effects may be related to down-regulation of Ki-67 and cyclin D1.

Key words: Breast neoplasms, Lysyl oxidase （LOX）, RNA interference, Cell proliferation, Proliferation factor