靶向抑制livin基因后结肠癌细胞对伊立替康敏感性的变化

Sensitivity changes of colon cancer cells to irinotecan after targeted restriction of livin gene expression

目的观察靶向抑制livin基因后结肠癌细胞HCT116对伊立替康敏感性的变化。方法采用脂质体包裹方法将载体介导的靶向livin基因的短发夹干扰RNA(a short hairpin RNA,shRNA)转入结肠癌细胞HCT116,48h后蛋白印迹方法检测RNA干扰的生物学效应,应用伊立替康0.01、0.1、1、10μg/mL梯度浓度作用于结肠癌细胞,24 h后MTT方法检测抑制livin基因表达后结肠癌细胞对伊立替康的敏感性变化,分光光度法检测结肠癌细胞内凋亡相关因子caspase-3活性的改变。结果靶向livin的shRNA有效地抑制了结肠癌细胞HCT116内livin基因的表达,抑制livin基因表达后结肠癌细胞对伊立替康的敏感性得到明显提高,细胞内的caspase-3活性显著增强。结论靶向抑制livin基因表达后能够激活细胞凋亡信号通路,增强伊立替康的抗结肠癌作用。

Objective. To explore sensitivity changes of colon cancer cells HCT116 to irinotecan after targeted restriction of livin gene expression. Methods A short hairpin RNA （shRNA） of livin gene in vector was transfected into colon cancer cells HCT116, the interference effect was evaluated by Western blot, HCT116 cells were exposed to irinotecan in 0. 01,0.1,1,10 μg/mL concentration, respectively, and sensitivity to irinotecan was evaluated by MTT, caspase-3 activity also was examined by colorimetric assay. Results Livin gene expression was effectively inhibited by targeted shRNA, sensitivity to irinotecan was increased after inhibition of livin expression, caspase-3 was also activated in tumor cells. Conclusion Apoptosis pathway can be activated by targeted restriction of livin expression, anti-tumor effect of irinotecan on colon cancer cells can be also increased.