摘要
客观:对 survivin 构造小介入 RNA (siRNA ) 的表示向量并且在 survivin 表示和人的胰腺的癌症房间线 PC-2 和乳癌房间线 MCF-7 的增长上观察它的效果。方法:对 survivin 和 transfected 构造了 siRNA 的表示向量它进用 lipofectamine 的 PC-2 和 MCF-7 房间 2000。survivin 的表示被 semi-quantitive RT-PCR 和免疫组织化学检测,并且它 PC-2 和 MCF-7 房间的增长上的效果被 MTT 试金检测。结果:顺序特定的 siRNA 的介绍能高效地在二根癌症房间线在 mRNA 和蛋白质层次压制 survivin 表示。在 PC-2 房间,表示抑制率在 mRNA 水平是 81.25% , 74.24% 在蛋白质铺平。在 MCF-7 房间,表示抑制率在 mRNA 水平是 64.91% , 79.72% 在蛋白质铺平。PC-2 和 MCF-7 房间的增长也被压制,并且在房间以后的 24 和 48 个小时被重新播种 PC-2 房间的增长抑制率是 28.00% 和 33.38% ,并且 MCF-7 房间的分别地是 31.58% 和 33.02% 。结论:对 survivin 的 siRNA 的表示向量能高效地并且明确地在 PC-2 和 MCF-7 房间堵住 survivin 表示。在 survivin 的规定下面,表示能压制 PC-2 和 MCF-7 房间的增长。Survivin RNAi 可以在人的癌症的基因治疗有潜在的价值。
Objective To construct an expression vector of small interfering RNA (siRNA) against survivin and observe its effects on survivin expression and proliferation of human pancreatic cancer cell line PC-2 and breast cancer cell line MCF-7.
Methods Constructed an expression vector of siRNA against survivin and transfected it into PC-2 and MCF-7 cells using lipofectamine^TM 2000. The expression of survivin was detected by semi-quanfifive RT-PCR and immunohistochemistry, and its effects on proliferation of PC-2 and MCF-7 cells were detected by MTT assay.
Results The introduction of sequence-specific siRNA could efficiently suppress survivin expression at both mRNA and protein levels in the two cancer cell lines. In PC-2 cells, the expression inhibition rates were 81.25% at mRNA level and 74.24% at protein level In MCF-7 cells, the expression inhibition rates were 64.91% at mRNA level and 79. 72% at protein level The proliferation of PC-2 and MCF-7 cells was also suppressed, and24 and 48 hours after the cells were reseeded, the proliferation inhibition rates of PC-2 cells were 28. 00% and 33. 38%, and that of MCF-7 cells were 31.58% and 33.02%, respectively.
Conclusions The expression vector of siRNA against survivin can block survivin expression in PC-2 and MCF-7 cells efficiently and specifically. Down regulation of survivin expression can suppress proliferation of PC-2 and MCF-7 cells. Survivin RNAi may have potential value in gene therapy of human cancers.
基金
Supported by the Key Science and Technology Research Project ofShaanxi Province [2003K10-G35,2004K13-G11(1)].
关键词
胰腺癌
乳腺癌
肿瘤细胞
细胞增殖
RNA interference
small interfering RNA
survivin
pancreatic cancer
breast cancer