Development of anti-cancer strategy to increase sensitivity of cancer cells to chemotherapy using siRNA combined with HVJ-E vector

利用siRNA联合HVJ-E载体开发提高癌细胞对化疗敏感性的抗癌策略

基本信息

批准号：
15300163
负责人：
KANEDA Yasufumi
金额：
$ 8.51万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

项目摘要

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 target protein by specifically digesting its mRNA. Suppression of the machineries using siRNA may enhance the sensitivity to chemotherapy in cancers when combined with effective delivery system. To enhance the anti-cancer effect of chemotherapy, we tested the ability of short interfering RNA(siRNA) to inhibit chemotherapy-resistant gene expression. We found that Rad51 was enhanced with cis-diamminedichloroplatinum (II) (CDDP ; cisplatin) in cancer cells. Overexpression of Rad51 caused cancer cells to become resistant to CDDP. When synthetic Rad 51 siRNA was delivered to HeLa cells using HVJ (hemagglutinating virus of Japan, Sendai virus) envelope vector, the delivery efficiency of siRNA was approximately 100%. No Rad51 transcripts were detected on day 2, and Rad51 protein completely disappeared for 4 days after siRNA transfer. The combination of Rad51 siRNA and CDDP caused increased DNA damage, which was detected by pulsed-field gel electrophoresis. When HeLa cells were incubated with 0.02 μg/ml CDDP for 3 hours after siRNA transfer, the number of colonies decreased to approximately 10% of that with scrambled siRNA. When Rad51 siRNA was delivered into tumors using HVJ envelope vector, the delivery efficiency was approximately 50%, and the Rad51 transcript level was reduced by approximately 60%. Rad51 siRNA combined with CDDP significantly inhibited the tumor growth when compared to siRNA or CDDP alone.Our results suggest that the combination of CDDP and Rad51 siRNA will be an effective anti-cancer protocol.

每种癌症疗法似乎都能暂时有效地消退癌症，但癌症也会逐渐产生抵抗化疗的机制。短干扰 RNA (siRNA) 已被评估为一种有吸引力且有效的抑制靶蛋白的工具。通过特异性消化其mRNA，当与有效的递送系统结合时，使用siRNA抑制该机制可能会增强癌症对化疗的敏感性。为了增强化疗的抗癌效果，我们测试了短干扰的能力。 RNA(siRNA) 抑制化疗耐药基因的表达。我们发现，顺式二氯铂 (II)（CDDP；顺铂）在癌细胞中过度表达 Rad51，导致癌细胞对 CDDP 产生耐药性。使用HVJ（日本血凝病毒，仙台病毒）包膜载体将siRNA递送至HeLa细胞，siRNA的递送效率约为100%。 Rad51 转录物在 siRNA 转移后 4 天检测到 Rad51 转录物，当 HeLa 细胞与 0.02 μg 一起孵育时，Rad51 siRNA 和 CDDP 的组合导致 DNA 损伤增加。 /ml CDDP siRNA 转移后 3 小时，集落数量减少至乱序 siRNA 时的集落数量约 10%。使用HVJ包膜载体，递送效率约为50％，并且与单独的siRNA或CDDP相比，Rad51 siRNA与CDDP的组合显着抑制了肿瘤生长。我们的结果表明，Rad51 siRNA与CDDP的组合显着抑制了肿瘤生长。 CDDP和Rad51 siRNA将是一种有效的抗癌方案。