Wip1基因沉默对结肠癌细胞化疗敏感性的影响

Effect of Wip1 gene silencing on chemotherapy sensitivity of human colon cancer cells

目的:观察靶向Wip1基因的特异性siRNA序列对结肠癌细胞Wip1基因的抑制效应,探讨Wip1基因沉默对结肠癌细胞化疗敏感性的影响。方法:将Wip1-811 siRNA转染入Wip1高表达的RKO结肠癌细胞株,采用real-time PCR法检测Wip1 mRNA的表达,采用Western blotting方法检测Wip1蛋白表达,采用MTS方法检测结肠癌细胞活力,流式细胞术检测细胞凋亡及细胞周期。结果:Wip1-811 siRNA明显抑制Wip1的表达。抑制Wip1基因表达后,转染组RKO结肠癌细胞对抗肿瘤药物的敏感性增加,5-氟尿嘧啶处理组RKO结肠癌细胞活力由(89.4±6.6)%降至(74.7±3.9)%(P<0.05),奥沙利铂处理组细胞活力由(77.9±2.4)%降至(66.7±2.9)%(P<0.05)。转染Wip1-811 siRNA后,5-氟尿嘧啶处理组细胞凋亡比例由(7.7±0.5)%升至(12.3±3.2)%(P<0.05);奥沙利铂处理组细胞凋亡比例由(14.7±2.1)%升至(34.0±2.1)%(P<0.05)。结论:沉默Wip1基因表达能增强结肠癌细胞的化疗敏感性。

AIM：To observe the inhibitory effect of siRNA targeting to Wip1 gene on the Wip1 gene expression in the colon cancer cells and to investigate the influence of Wip1 gene silencing on the chemotherapy sensitivity of colon cancer cells.METHODS：Wip1-811 siRNA targeting to Wip1 gene was transfected into RKO colon cancer cells with high expression of Wip1 gene.The mRNA expression of Wip1 was measured by real-time PCR.The protein level of Wip1 was detected by Western blotting.The viability of RKO colon cancer cells was measured by MTS assay.The cell apoptosis and cell cycle were analyzed by flow cytometry.RESULTS： Wip1-811 siRNA efficiently inhibited the expression of Wip1 at mRNA and protein levels.The enhanced chemotherapy sensitivity of RKO colon cancer cells was observed after inhibition of Wip1 gene expression.The viability of RKO colon cancer cells was decreased from （89.4 ±6.6）%to （74.7 ±3.9）%af-ter treated with 5-fluorouracil （P〈0.05） and decreased from （77.9 ±2.4）%to （66.7 ±2.9）%after treated with oxali-platin （ P〈0.05 ） .The cell apoptotic rate was increased from （ 7.7 ±0.5 ）% to （ 12.3 ±3.2 ）% and from （ 14.7 ± 2.1）% to （34.0 ±2.1）% when RKO colon cancer cells were treated with 5-fluorouracil and oxaliplatin, respectively （P〈0.05）.CONCLUSION：Wip1 gene silencing enhances chemotherapy sensitivity of colon cancer cells.