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目的:在肿瘤侵袭转移的细胞和生物学机制中,包含有多种基因和信号传导通路的改变。PDK1是PI3K分子通路上的一个重要分子,为了研究PDK1对乳腺肿瘤细胞侵袭能力的影响,我们以乳腺癌MDA-MB-231细胞为研究模型,用小RNA干扰法特异下调PDK1的表达,研究其侵袭能力的变化,为乳腺癌临床治疗探索新的分子靶点。方法:通过构建PDK1特异的小RNA干扰质粒,利用脂质体介导的方法转染乳腺癌细胞MDA-MB-231,无限稀释法筛选单克隆细胞系,并通过Western Blot的方法检测各组细胞中PDK1蛋白的表达水平,细胞计数的方法观察各组细胞之间的增殖情况,并利用Matrigel Transwell的实验方法研究细胞在体外的侵袭能力差别。结果:MDA-MB-231细胞经小RNA干扰后PDK1蛋白表达水平明显下降,与对照组相比,细胞的生长增殖能力没有显著差异(P>0.05),细胞的侵袭能力显著下降,并且差异有统计学意义(P<0.05)。结论:MDA-MB-231是一种侵袭能力高的乳腺癌细胞系,本研究利用小RNA干扰技术,特异性下调了乳腺癌MDA-MB-231细胞中的PDK1蛋白表达,结果表明,乳腺癌细胞的生长能力与对照组的细胞相比没有明显差别,但是,PDK1蛋白表达水平下降后乳腺癌细胞的侵袭能力明显下降,提示我们PDK1可能与乳腺癌MDA-MB-231细胞的侵袭能力肾密相关。鉴于PDK1处于PDK1-Akt-PKC信号通路的上游,以PDK1为靶标的分子干预可能对乳腺癌细胞的侵袭转移起到有效的作用。
OBJECTIVE: To investigate the cellular and biological mechanisms involved in the invasion and metastasis of tumors, including the alteration of multiple genes and signaling pathways. PDK1 is an important molecule in the PI3K molecular pathway. In order to study the effect of PDK1 on the invasiveness of breast tumor cells, we used a small RNA interference method to specifically downregulate the expression of PDK1 in breast cancer MDA-MB-231 cells, Invasive ability of breast cancer, explore new molecular targets for clinical treatment. METHODS: PDK1-specific small interfering plasmid was constructed and transfected into breast cancer cell MDA-MB-231 by liposome-mediated method. The monoclonal cell lines were screened by immiscible dilution and the cells were detected by Western Blot In PDK1 protein expression levels, cell counting method to observe the proliferation of cells between groups, and the use of Matrigel Transwell experimental method of cell invasion in vitro differences. Results: The expression of PDK1 protein in MDA-MB-231 cells was significantly decreased after small RNA interference. Compared with the control group, the proliferation and proliferation ability of MDA-MB-231 cells showed no significant difference (P> 0.05) Statistical significance (P <0.05). Conclusion: MDA-MB-231 is a highly invasive breast cancer cell line. In this study, we used a small RNA interference technology to specifically down-regulate PDK1 protein expression in breast cancer MDA-MB-231 cells. However, the decrease of PDK1 protein expression in invasive breast cancer cells significantly decreased, suggesting that our PDK1 may be associated with the invasiveness of breast cancer MDA-MB-231 cells kidney Related. Because PDK1 is upstream of PDK1-Akt-PKC signaling pathway, PDK1-targeted molecular intervention may play an effective role in the invasion and metastasis of breast cancer cells.