| 牛朝霞,彭蕤蕤,陈洁,等.靶向survivin基因的小干扰RNA对食管癌细胞Eca-109 化疗敏感性的影响[J].安徽医药,2016,20(4):683-686. |
| 靶向survivin基因的小干扰RNA对食管癌细胞Eca-109 化疗敏感性的影响 |
| Effects of siRNA targeting survivin gene on chemosensitivityof esophageal carcinoma cell Eca-109 |
| 投稿时间：2016-01-15 |
| DOI： |
| 中文关键词: 小干扰RNA survivin基因 Eca-109细胞 化疗敏感性 |
| 英文关键词: small interference RNA survivin gene Eca-109 cell chemosensitivity |
| 基金项目:河南省教育厅科学技术研究重点项目(No 14B310002)靶向survivin基因的小干扰RNA对食管癌细胞Eca-109 化疗敏感性的影响牛朝霞,彭蕤蕤,陈洁,秦紫芳,裴瑞 (河南医学高等专科学校病理生理学教研室,河南 郑州 451191)〗靶向survivin基因的小干扰RNA对食管癌细胞Eca-109 化疗敏感性的影响牛朝霞,彭蕤蕤,陈洁,秦紫芳 |
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| 中文摘要: |
| 目的 利用RNAi (RNA interference, RNAi) 技术抑制食管癌Eca-109细胞survivin基因表达,观察survivin基因沉默后对Eca-109细胞化疗敏感性的影响。方法 构建靶向survivin基因特定序列的小干扰RNA(small interference RNA, siRNA)真核表达载体并转染Eca-109细胞,筛选得到稳定转染的survivin干扰组细胞Eca-109 /si-survivin,同时设转染无关干扰质粒的Eca-109细胞为阴性对照组,未转染的Eca-109细胞为空白对照组；通过Western Blot法检测干扰前后survivin基因沉默抑制效果；随后将各组细胞与不同浓度的氟尿嘧啶(5-Fu)作用,MTT法检测各组细胞对5-Fu的半数抑制浓度(IC50),流式细胞仪分析各组细胞的凋亡率。结果 干扰组Eca-109 /si-survivin细胞survivin蛋白表达水平(18.75±3.12)较阴性对照组Eca-109 /si-control(44.17±3.15)及空白对照组Eca-109 (46.20±2.62)明显下调(P<0.05)；联合应用5-Fu的Eca-109 /si-survivin干扰组细胞IC50为(18.75±1.53)mg·L-1,显著低于空白对照组(39.65±1.75)mg·L-1和阴性对照组(38.95±1.34)mg·L-1,差异具有统计学意义(P<0.05)；联合应用5-Fu的Eca-109 /si-survivin干扰组细胞凋亡率为(37.35±1.52)%,明显高于空白对照组(11.26±1.21)%和阴性对照组(12.34±1.32)%,差异具有显著性(P<0. 05)。 结论 靶向survivin的siRNA能特异性沉默survivin基因表达,抑制细胞增殖,诱导细胞凋亡,并增强人食管癌Eca-109细胞对5-Fu的化疗敏感性。 |
| 英文摘要: |
| Objective To inhibit survivin gene expression by RNA interference technology and investigate the effects on chemosensitivity of esophageal carcinoma cell Eca-109. Methods siRNA eukaryotic expressing vector targeting specific sequence of survivin gene was constructed to transfect Eca-109 cells which were screened to select the survivin interference group Eca-109 /si-survivin with stable transfection.Meanwhile, the Eca-109 cells transfected with non-sense sequence were used as the negative control group and those untransfected were used as the blank control group.The expression level of survivin protein was detected by western blot to observe the interference effect. Then the cells of each group were treated with different concentrations of fluorouracil (5-Fu) , IC50 of 5-Fu in each group was detected by MTT and cell apoptosis rate was analyzed by flow cytometry. Results The survivin protein level (18.75±3.12) of the interference group Eca-109 /si-survivin was effectively lower than that of the negative control group Eca-109 /si-control(44.17±3.15)and the blank control group Eca-109 (46.20±2.62) (P<0.05); IC50 of 5-Fu in the interference group Eca-109 /si-survivin was(18.75±1.53)mg·L-1 , which was markedly lower than that of the blank control group(39.65±1.75)mg·L-1 and the negative control group(38.95±1.34)mg·L-1 with a statistically significant difference (P<0.05). The cell apoptosis rate of 5-Fu in the interference group Eca-109 /si-survivin was(37.35±1.52)%, which was evidently higher than that of the blank control group(11.26±1.21)% and the negative control group(12.34±1.32)% with a statistically significant difference (P<0.05). ConclusionssiRNA targeting survivin can specifically silence the expression of survivin gene, inhibit cell proliferation, induce cell apoptosis, and thus enhance the chemosensitivity of human esophageal cancer Eca-109 cells to 5-Fu. |
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