RNAi对骨肉瘤MG-63细胞survivin基因表达抑制作用

The gene silencing mediated by RNAi on survivin expression in osteosarcoma cell line MG-63

背景与目的:骨肉瘤是青少年中最常见的恶性肿瘤,它恶性程度高,预后差。survivin基因是作用最强的凋亡抑制因子,其表达与肿瘤的恶性程度及患者预后密切相关。本实验应用RNA干扰(RNA interfering,RNAi)技术研究特异性survivin-siRNA对骨肉瘤MG-63细胞survivin基因表达的干扰作用,并对RNAi技术在骨肉瘤基因治疗领域中应用的可能性进行初步分析。方法:将化学合成的特异性survivin-siRNA1和特异性survivin-siRNA2在脂质体的介导下转染骨肉瘤MG-63细胞,应用逆转录聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)和流式细胞术检测转染前后MG-63细胞中survivin-mRNA和蛋白水平,通过四甲基偶氮唑蓝(MTT)法来评估特异性survivin-siRNA对骨肉瘤MG-63细胞的抗增殖作用,观察MG-63细胞生物学特性的改变。结果:特异性survivin-siRNA降低了骨肉瘤MG-63细胞survivin mRNA和蛋白的表达水平,干涉后细胞的增殖受到抑制。结论:化学合成的survivin-siRNA能有效抑制survivin基因在骨肉瘤MG-63细胞中的表达,并对骨肉瘤MG-63细胞增殖有抑制作用。

Background and purpose： Osteosarcoma is the most common malignancies with high degree of malignancy and poor prognosis in young people. Survivin is a member of the inhibitor of apoptosis （lAP） protein gene family and its expression is related to the degree of malignancy and prognosis in osteosarcoma. The experiment was to study the inhibition of expression of survivin gene by synthesized short survivin-siRNA in osteosarcoma cell line MG-63. Methods： Chemically synthesized short survivin-siRNA was transfected into the osteosarcoma cell line MG-63. The survivin mRNA and protein level were detected by reverse transcription-polymerase chain reaction （RT-PCR） and flow cytometry （FCM）, the biological morphology and growth inhibition of MG-63 were observed with light microscopy and MTY assay, respectively. Results： Short siRNA targeting survivin down-regulated the transcription of survivin oncogene and protein level. The proliferation of osteosarcoma cell line MG-63 was inhibited after transfection. Conclusions： Chemically synthesized short survivin-siRNA can effectively inhibit survivin expression and cell proliferation in osteosarcoma cell line MG-63. Short survivin-siRNA may be a useful therapeutic strategy for cancer through the regulation of gene expression.