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RNA干扰介导的生存素下调抑制人胃癌细胞生长。

Downregulation of survivin by RNAi inhibits growth of human gastric carcinoma cells.

作者信息

Miao Guo-Ying, Lu Qi-Ming, Zhang Xiu-Lan

机构信息

Department of Digestion, The People's Hospital of Gansu Province, Lanzhou 730000, Gansu Province, China.

出版信息

World J Gastroenterol. 2007 Feb 28;13(8):1170-4. doi: 10.3748/wjg.v13.i8.1170.

Abstract

AIM

To investigate the inhibitory effect of a specific small survivin interfering RNA (siRNA) on cell proliferation and the expression of survivin in human gastric carcinoma cell line SGC-7901.

METHODS

To knockdown survivin expression, a small interfering RNA targeting against survivin was synthesized and transfected into SGC-7901 cells with lipofectamine 2000. The downregulation of survivin expression at both mRNA and protein levels were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis. Cell proliferation inhibition rates were determined by methyl thiazolyl tetrazolium (MTT) assay. The effect of survivin siRNA on cell cycle distribution and cell apoptosis was determined by flow cytometry (FCM).

RESULTS

RNA interference could efficiently suppress the survivin expression in SGC-7901 cells. At 48 h after transfection, the expression inhibition rate was 44.52% at mRNA level detected by RT-PCR and 40.17% at protein level by Western blot analysis. Downregulation of survivin resulted in significant inhibition of tumor cell growth in vitro. The cell proliferation inhibition rates at 24, 48 and 72 h after survivin siRNA and non-silencing siRNA transfection, were 34.06%, 47.61% and 40.36%, respectively. The apoptosis rate was 3.56% and the number of cells was increased in G(0)/G(1) phase from 38.2% to 88.6%, and decreased in S and G(2)/M phase at 48 h after transfection.

CONCLUSION

Downregulation of survivin results in significant inhibition of tumor growth in vitro. The inhibition of survivin expression can induce apoptosis of SGC-7901 cells. The use of survivin siRNA deserves further investigation as a novel approach to cancer therapy.

摘要

目的

研究特异性小干扰RNA(siRNA)对人胃癌细胞系SGC-7901细胞增殖及生存素(survivin)表达的抑制作用。

方法

合成针对survivin的小干扰RNA,用脂质体2000转染SGC-7901细胞以敲低survivin表达。通过逆转录聚合酶链反应(RT-PCR)和蛋白质印迹分析检测mRNA和蛋白质水平上survivin表达的下调情况。采用甲基噻唑基四氮唑(MTT)法测定细胞增殖抑制率。通过流式细胞术(FCM)确定survivin siRNA对细胞周期分布和细胞凋亡的影响。

结果

RNA干扰可有效抑制SGC-7901细胞中survivin的表达。转染后48小时,RT-PCR检测到mRNA水平的表达抑制率为44.52%,蛋白质印迹分析检测到蛋白质水平的表达抑制率为40.17%。survivin的下调导致体外肿瘤细胞生长受到显著抑制。survivin siRNA和非沉默siRNA转染后24、48和72小时的细胞增殖抑制率分别为34.06%、47.61%和 40.36%。转染后48小时,凋亡率为3.56%,G(0)/G(1)期细胞数量从38.2%增加到88. 6%,S期和G(2)/M期细胞数量减少。

结论

survivin的下调导致体外肿瘤生长受到显著抑制。survivin表达的抑制可诱导SGC-7901细胞凋亡。作为一种新的癌症治疗方法,survivin siRNA的应用值得进一步研究。

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