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Traf6通过降低TAK1活性抑制乳腺癌MCF 7细胞增殖能力
米慧琼,罗传文,于力,张少立,张英
0
(恩施土家族苗族自治州中心医院1.肿瘤放疗中心;恩施土家族苗族自治州中心医院核医学科)
摘要:
【摘要】 目的 探讨肿瘤坏死因子相关受体因子6(Traf6)对乳腺癌MCF7细胞增殖的影响及其可能的作用机制。方法 采用pGPU6/Neo载体构建稳定敲除Traf6的MCF7细胞株,应用实时定量PCR和免疫印迹验证Traf6基因和蛋白的均低表达,采用细胞计数试剂盒8(CCK8)法检测MCF7细胞增殖情况,流式细胞仪检测MCF7细胞周期的变化,蛋白印记法(Western Blot)检测MCF7细胞中周期蛋白A(Cyclin A)、周期蛋白B(Cyclin B)、周期蛋白依赖激酶(CDK/p34)表达量和转录生长因子β活化激酶1(TAK1)磷酸化水平。结果 使用pGPU6/Neo载体能够稳定的敲除MCF7细胞中Traf6的表达,敲除Traf6能够抑制MCF7细胞增殖并诱导其凋亡,将细胞阻滞在G1期,同时抑制细胞周期蛋白Cyclin A、Cyclin B及CDK/p34的表达及TAK1磷酸化。结论 Traf6表达的降低能抑制TAK1信号通路活性,进而抑制乳腺癌MCF7细胞增殖能力。
关键词:  乳腺癌细胞  肿瘤坏死因子相关受体因子6  TAK1  敲除  增殖  凋亡
DOI:
基金项目:
Tumor necrosis factor related receptor factor 6 suppresses proliferation of breast cancer MCF 7 cells via inhibiting the TAK1 activity
MI Huiqiong,LUO Chuanwen,YU Li,ZHANG Shaoli,ZHANG Ying
(Tumor Radiotherapy Center, Central Hospital of Enshi Autonomous Prefecture;Department of Nuclear Medicine, Central Hospital of Enshi Autonomous Prefecture)
Abstract:
【Abstract】 Objective To study the effect of tumor necrosis factor related receptor factor 6 (Traf6) on the proliferation of breast cancer MCF7 cells and its possible mechanism. Methods The qualified recombinant plasmid shTraf6pGPU6/Neo was transfected into MCF7 cells to knock down Traf6 related gene. The level of Traf6 was detected on both mRNA and protein levels by realtime PCR and Western blot. CCK8 assay and flow cytometry were performed to detect the change of proliferation and cell cycle in MCF7 cells. The expression of cell cycle associated proteins (Cyclin A, Cyclin B and CDK/p34) and TAK phosphorylation were also detected and recorded. Results The Traf6 plasmid was constructed successfully. MCF7 cells with Traf6 knocking down had been established successfully. Knocking down of Traf6 inhibited the proliferation of MCF7 significantly, stimulated the apoptosis of MCF7 cells, and cells were blocked in G1phase. Moreover, the Traf6 knock out increased the expression levels of cycle proteins (Cyclin A, Cyclin B and CDK/p34) and TAK1 phosphorylation. Conclusion Suppression of Traf6 can inhibit the proliferation and induce the apoptosis of MCF7 cells through TAK1 signaling pathway.
Key words:  Breast cancer cells  Tumor necrosis factor related receptor factor 6  TAK1  Knock down  Proliferation  Apoptosis

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