联合靶向mTOR和Gli1/2信号通路的药物对肾癌细胞生长的抑制效应

目的:探讨联合靶向mTOR和Gli1/2信号通路的药物对肾癌细胞生长的抑制效应。方法:采用Real time-PCR的方法检测SHH/Gli信号通路的核心效应成分Gli1和Gli2在20对正常肾脏和肾癌组织中的表达情况；采用Western blot技术验证4对正常肾脏和配对的肿瘤中Gli1和Gli2蛋白的表达水平。单独或联合使用不同浓度的mTOR抑制剂与Gli1/2抑制剂于肾癌细胞后，采用MTT的方法检测细胞的体外增殖能力，然后采用Western blot技术检测联合应用低剂量mTOR抑制剂与Gli1/2抑制剂的肾癌细胞内细胞周期、凋亡相关蛋白的变化情况。结果:Gli1与Gli2在肾癌组织中的表达高于对照的正常肾脏组织。低剂量（5 μmol/L）Gli1/2抑制剂Gant61并不能显著抑制肾癌细胞的生长，高剂量（10 μmol/L）的Gant61能达到有效的抑制效应；但5 μmol/L Gant61与低剂量mTOR抑制剂Rapamycin联用时，肾癌细胞的生长显著被抑制，且诱导细胞凋亡，阻滞细胞周期。结论:联合靶向mTOR和Gli1/2信号通路显著抑制肾癌细胞生长，为潜在的治疗策略。

The suppressive effect of the combination treatment of targeting mTOR and Gli1/2 signaling on cell growth in renal cell carcinoma cells

Objective:To explore the suppressive effect of combinatory targeted therapies of mTOR and Gli1/2 signaling on RCC cells.Methods:Real time-PCR and Western blot were used to detect the expression of critical component of SHH/Gli pathway，Gli1 and Gli2 in 20 paired human RCC and normal tissues.RCC cells were treated with single or combinatory mTOR inhibitor and Gli1/2 inhibitor，and MTT assays were used to detect cell proliferation.Western blot and then was used to detect cell cycle an apoptosis marker levels in cells treated.Results:The expression of Gli1 and Gli2 were increased in RCC tissues compared to normal tissues.High dose（10 μmol/L） but not low dose（5 μmol/L） of Gli1/2 inhibitor Gant61 could significantly inhibit RCC growth.When low dose of Gant61 was combinatory used with mTOR inhibitor Rapamycin，cell growth was significantly suppressed，the cell cycle in cells was blocked and apoptosis was induced. Conclusion:Simultaneously targeting mTOR and Gli1/2 signaling would be potential treatment for RCC patients clinically.