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Down-regulation of survivin expression by small interfering RNA induces pancreatic cancer cell apoptosis and enhances its radiosensitivity

机译:小干扰RNA下调survivin的表达诱导胰腺癌细胞凋亡并增强其放射敏感性

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摘要

AIM: To investigate the inhibitory effect of small interfering RNA (siRNA) on the expression of survivin in pancreatic cancer cell line PC-2 and the role of siRNA in inducing PC-2 cell apoptosis and enhancing its radiosensitivity.METHODS: A siRNA plasmid expression vector against survivin was constructed and transfected into PC-2 cells with LipofectamineTM 2000. The down regulation of survivin expression was detected by semi-quantitive RT-PCR and immunohistochemical SP method and the role of siRNA in inducing PC-2 cell apoptosis and enhancing its radiosensitivity was detected by flow cytometry.RESULTS: The sequence-specific siRNA efficiently and specifically down-regulated the expression of survivin at both mRNA and protein levels. The expression inhibition ratio was 81.25% at mRNA level detected by semi-quantitive RT-PCR and 74.24% at protein level detected by immunohistochemical method. Forty-eight hours after transfection,apoptosis was induced in 7.03% cells by siRNA and in 14.58% cells by siRNA combined with radiation.CONCLUSION: The siRNA plasmid expression vector against survivin can inhibit the expression of survivin in PC-2 cells efficiently and specifically. Inhibiting the expression of survivin can induce apoptosis of PC-2 cells and enhance its radiosensitivity significantly. RNAi against survivin is of potential value in gene therapy of pancreatic cancer.
机译:目的:探讨小干扰RNA(siRNA)对胰腺癌细胞株PC-2中survivin表达的抑制作用,以及siRNA在诱导PC-2细胞凋亡和增强放射敏感性中的作用。构建了针对survivin的载体,并用Lipofectamine TM 2000将其转染到PC-2细胞中。通过半定量RT-PCR和免疫组化SP方法检测了survivin表达的下调以及siRNA在诱导中的作用。结果:流式细胞仪检测PC-2细胞凋亡并增强其放射敏感性。结果:序列特异性siRNA在mRNA和蛋白质水平上均有效且特异性地下调了survivin的表达。通过半定量RT-PCR检测到的mRNA水平的表达抑制率为81.25%,通过免疫组织化学法检测到的蛋白水平的表达抑制率为74.24%。转染后48小时,siRNA联合放射诱导siRNA诱导7.03%的细胞凋亡,siRNA联合放射诱导14.58%的细胞凋亡。结论:针对survivin的siRNA质粒表达载体可以有效,特异性地抑制survivin在PC-2细胞中的表达。 。抑制survivin的表达可诱导PC-2细胞凋亡,并显着增强其放射敏感性。针对存活蛋白的RNAi在胰腺癌的基因治疗中具有潜在价值。

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