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背景与目的存活素特异地在肿瘤组织中过表达,许多报道表明抑制它的功能对肿瘤治疗有利。RNA干扰被证明是一项能有效而特异地抑制基因表达的新技术。本研究的目的在于构建存活素基因的小分子干扰RNA(siRNA)表达质粒,并检测该质粒在乳腺癌细胞中的生物学功能。方法应用含有U6启动子的mU6pro载体构建存活素siRNA质粒,RT-PCR和Westernblot法检测该质粒转染MCF-7细胞后存活素表达的变化,流式细胞仪和MTT法分别检测其对细胞周期和对细胞增殖的影响。结果存活素siRNA质粒明显下调MCF-7细胞中存活素的表达,阻断细胞周期在G1期,显著抑制细胞增殖,促进细胞凋亡。结论通过RNA干扰,存活素的表达在mRNA和蛋白质水平上被下调。
BACKGROUND & OBJECTIVE Survivin is overexpressed specifically in tumor tissue and many reports indicate that inhibition of its function is beneficial for cancer therapy. RNA interference proved to be a new technology that can effectively and specifically inhibit gene expression. The purpose of this study was to construct a small interfering RNA (siRNA) expression plasmid for survivin gene and to examine the biological function of the plasmid in breast cancer cells. Methods Survivin siRNA plasmid was constructed by mU6pro vector containing U6 promoter. The expression of survivin was detected by RT-PCR and Western blotting. The expression of survivin was detected by MTT assay and flow cytometry And the impact on cell proliferation. Results survivin siRNA plasmid significantly down-regulated the expression of survivin in MCF-7 cells, blocking the cell cycle in G1 phase, significantly inhibiting cell proliferation and promoting apoptosis. Conclusions Survivin expression is down-regulated at the mRNA and protein levels by RNA interference.