Rad51 siRNA delivered by HVJ envelope vector enhances the anti-cancer effect of cisplatin
Rad51 siRNA delivered by HVJ envelope vector enhances the anti-cancer effect of cisplatin
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DOI：
10.1002/jgm.753
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发表时间：
2005-08-01
影响因子：
3.5
通讯作者：
Kaneda, Y
中科院分区：
文献类型：
--
作者：
Ito, M;Yamamoto, S;Kaneda, Y
背景每一种癌症治疗似乎对癌症消退都是短暂有效的，但癌症会逐渐转变为对治疗产生耐药性。癌症也会发展出抵抗化疗的机制。短干扰RNA（siRNA）已被评估为通过特异性消化其mRNA来抑制靶蛋白的有吸引力的和有效的工具。使用siRNA的机制的抑制可能会提高癌症化疗的敏感性时，结合一个有效的delivery system.Methods为了提高化疗的抗癌效果，我们转移到各种人类癌细胞使用HVJ（日本血凝病毒，仙台病毒）包膜载体的存在或不存在的顺二氨二氯铂（II）（CDDP，顺铂）的siRNA对Rad 51。在膜联蛋白V标记后，通过改良的MTT测定、细胞计数或荧光激活细胞分选（FACS）分析来评估细胞生长的抑制。结果用HVJ包膜载体将合成的Rad 51 siRNA导入HeLa细胞后，第2天检测不到Rad 51的转录产物，转染后第4天Rad 51蛋白完全消失。当HeLa细胞在siRNA转移后与0.02 μ g/ml CDDP孵育3小时时，集落数减少到用乱序siRNA的集落数的约10%。对CDDP的敏感性在各种人类癌细胞中增强，但在正常人类成纤维细胞中不增强。当使用HVJ包膜载体将Rad 51 siRNA递送到肿瘤中时，Rad 51转录物水平降低至约25%。结论Rad 51 siRNA在体内外均能增强肿瘤细胞对顺铂的敏感性。我们的研究结果表明，CDDP和Rad 51 siRNA的组合将是一种有效的抗癌方案。版权所有（c）2005年约翰威利父子有限公司。
Background Every cancer therapy appears to be transiently effective for cancer regression, but cancers gradually transform to be resistant to the therapy. Cancers also develop machineries to resist chemotherapy. Short interfering RNA (siRNA) has been evaluated as an attractive and effective tool for suppressing a target protein by specifically digesting its mRNA. Suppression of the machineries using siRNA may enhance the sensitivity to chemotherapy in cancers when combined with an effective delivery system.Methods To enhance the anti-cancer effect of chemotherapy, we transferred siRNA against Rad51 into various human cancer cells using the HVJ (hemagglutinating virus of Japan, Sendai virus) envelope vector in the presence or absence of cis-diamminedichloroplatinum(II) (CDDP, cisplatin). The inhibition of cell growth was assessed by a modified MTT assay, counting cell number, or fluorescence-activated cell sorting (FACS) analysis after Annexin V labeling. The synthetic Rad51 siRNA was also introduced into subcutaneous tumor masses of HeLa cells in SCID mice with or without intraperitoneal injection of CDDP, and tumor growth was monitored.Results When synthetic Rad51 siRNA was delivered into HeLa cells using the HVJ envelope vector, no Rad51 transcripts were detected on day 2, and Rad51 protein completely disappeared for 4 days after siRNA transfer. When HeLa cells were incubated with 0.02 mu g/ml CDDP for 3 h after siRNA transfer, the number of colonies decreased to approximately 10% of that with scrambled siRNA. The sensitivity to CDDP was enhanced in various human cancer cells, but not in normal human fibroblasts. When Rad51 siRNA was delivered into tumors using the HVJ envelope vector, the Rad51 transcript level was reduced to approximately 25%. Rad51 siRNA combined with CDDP significantly inhibited tumor growth when compared to siRNA or CDDP alone.Conclusions Rad51 siRNA could enhance the sensitivity to CDDP in cancer cells both in vitro and in vivo. Our results suggest that the combination of CDDP and Rad51 siRNA will be an effective anti-cancer protocol. Copyright (c) 2005 John Wiley & Sons, Ltd.