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目的:应用RNA i的方法抑制丝氨酸蛋白酶LON表达,研究该丝氨酸蛋白酶的表达与肿瘤增殖以及药物敏感性之间的关系。方法:以人乳腺癌细胞MCF-7为研究对象,通过RNA i下调LON基因表达,采用RT-PCR法、MTT增殖实验、流式细胞术等技术进行检测。结果:在瞬时转染与稳定转染细胞株中,RT-PCR结果证实LON基因的表达明显下调;MTT增殖实验结果显示,LON基因的下调可导致细胞的增殖能力降低,并且能够增加对化疗药顺铂的药物敏感性。流式细胞仪检测显示LON基因的下调对细胞周期没有显著影响。结论:通过RNA i下调LON基因能够抑制肿瘤细胞的增殖并且与顺铂有协同作用,其抑制作用未造成细胞周期的改变。
OBJECTIVE: To study the relationship between the serine protease expression and tumor proliferation and drug sensitivity by RNAi method. Methods: The human breast cancer cell line MCF-7 was used as the research object. The gene expression of LON was down-regulated by RNAi. The expression of LON gene was detected by RT-PCR, MTT assay and flow cytometry. Results: RT-PCR results showed that LON gene expression was significantly down-regulated in transiently transfected and stably transfected cell lines; MTT proliferation assay showed that down-regulation of LON gene led to the decrease of cell proliferation ability, Cisplatin drug sensitivity. Flow cytometry showed that downregulation of LON gene had no significant effect on cell cycle. CONCLUSION: Down-regulation of LON gene by RNAi can inhibit the proliferation of tumor cells and has a synergistic effect with cisplatin, and its inhibition does not cause cell cycle changes.