Down-regulation of survivin expression by small interfering RNA induces pancreatic cancer cell apoptosis and enhances its radiosensitivity
Down-regulation of survivin expression by small interfering RNA induces pancreatic cancer cell apoptosis and enhances its radiosensitivity
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DOI：
10.3748/wjg.v12.i18.2901
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发表时间：
2006-05-14
影响因子：
4.3
通讯作者：
Zhao-Yin Qin
中科院分区：
文献类型：
--
作者：
Hai-Tao Guan;Xing-Huan Xue;Zhao-Yin Qin
目的：探讨小干扰RNA（siRNA）对胰腺癌细胞系PC-2中survivin表达的抑制作用以及siRNA诱导PC-2细胞凋亡、增强放射敏感性的作用。方法：构建survivin siRNA质粒表达载体，用Lipofectamine(TM) 2000转染PC-2细胞，观察其下调survivin表达的情况。 采用半定量RT-PCR和免疫组化SP法检测，流式细胞术检测siRNA诱导PC-2细胞凋亡和增强放射敏感性的作用。结果：序列特异性siRNA在mRNA和蛋白水平上高效、特异地下调survivin的表达。半定量RT-PCR检测mRNA水平表达抑制率为81.25%，免疫组化法检测蛋白水平表达抑制率为74.24%。转染48小时后，siRNA诱导7.03%的细胞凋亡，siRNA联合辐射诱导14.58%的细胞凋亡。结论：针对survivin的siRNA质粒表达载体能够高效、特异地抑制PC-2细胞中survivin的表达。抑制survivin表达可诱导PC-2细胞凋亡，显着增强其放射敏感性。针对生存素的 RNAi 在胰腺癌的基因治疗中具有潜在价值。 (C) 2006 年 WJG 出版社。版权所有。
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 Lipofectamine(TM) 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. (C) 2006 The WJG Press. All rights reserved.