澳洲茄碱通过mTOR信号通路抑制膀胱癌细胞增殖

Solasonine inhibits the proliferation of bladder cancer cells through mTOR signaling pathway

目的研究澳洲茄碱对膀胱癌细胞增殖、细胞凋亡、细胞周期的影响及其作用机制。方法0、5、10、20、40μmol/L澳洲茄碱处理膀胱癌J82和T24细胞24 h、48 h、72 h后,应用CCK-8检测细胞生存率。0、10、30μmol/L澳洲茄碱处理J82细胞24 h后,用流式细胞术检测细胞凋亡和细胞周期。0、10、30μmol/L澳洲茄碱处理J82细胞24 h后,应用Western blot法检测mTOR、p-mTOR、p70S6K、p-p70S6K蛋白的表达。0、10、20、30μmol/L澳洲茄碱处理J82细胞24 h后,RT-qPCR法检测mTOR、p70S6K mRNA表达情况。结果澳洲茄碱显著抑制膀胱癌J82细胞、T24细胞增殖,相同作用时间、不同浓度组细胞生存率总体差异均有统计学意义(P<0.01)。澳洲茄碱诱导J82细胞凋亡,0、10、30μmol/L澳洲茄碱组凋亡率分别为(0.93±0.26)%、(8.33±2.16)%、(29.53±7.63)%。澳洲茄碱引起J82细胞发生S期阻滞,0、10、30μmol/L澳洲茄碱组S期细胞占比分别是(34.51±3.61)%、(39.34±3.21)%、(44.50±4.29)%。澳洲茄碱下调J82细胞p-mTOR蛋白表达(F=85.18,P<0.01)和p-p70S6K蛋白表达(F=74.96,P<0.01)。澳洲茄碱下调J82细胞mTOR mRNA表达(F=31.49,P<0.01)和p70S6K mRNA表达(F=47.08,P<0.01)。结论澳洲茄碱通过mTOR信号通路抑制膀胱癌细胞增殖、促进细胞凋亡、阻滞细胞周期。

Objective To study the effect of solasonine on the proliferation,apoptosis and cell cycle of bladder cancer cells and its mechanism.Methods After the treatment of bladder cancer cells J82 and T24 with 0,5,10,20 and 40μmol/L solasonine for 24 h,48 h and 72 h,CCK-8 was used to detect cell proliferation.After the treatment of J82 cells with 0,10 and 30μmol/L solasonine for 24 h,FACS was performed to detect cell cycle and apoptosis.After the treatment of J82 cells with 0,10,20 and 30μmol/L solasonine for 24 h,Western blot was undergone to detect the expression of mTOR,p-mTOR,p70S6K and p-p70S6K protein.After the treatment of J82 cells with 0,10 and 30μmol/L solasonine for 24 h,RT-qPCR was used to detect the expression of mTOR and p70S6K mRNA.Results Solasonine significantly inhibited the proliferation of bladder cancer cell J82 and T24 cells(P<0.01).Solasonine induced apoptosis of J82 cells,the apoptosis rates of 0,10 and 30μmol/L solasonine groups were(0.93±0.26)%,(8.33±2.16)%and(29.53±7.63)%,respectively.Solasonine arrested J82 cells at S stage,the proportions of S stage of 0,10 and 30μmol/L solasonine groups were(34.51±3.61)%,(39.34±3.21)%and(44.50±4.29)%,respectively.Solasonine downregulated the protein expression of p-mTOR of J82 cells(F=85.18,P<0.01).Solasonine downregulated the protein expression of p-p70S6K of J82 cells(F=74.96,P<0.01).Solasonine downregulated the mRNA expression of mTOR of J82 cells(F=31.49,P<0.01).Solasonine downregulated the mRNA expression of p70S6K of J82 cells(F=47.08,P<0.01).Conclusion Solasonine can inhibit the proliferation of bladder cancer cells,induce cell apoptosis and arrest cell cycle through mTOR signaling pathway.