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目的:探讨氧化苦参碱对皮肤鳞状细胞癌(cSCC)Colo-16细胞株凋亡及自噬的影响.方法:用不同浓度的氧化苦参碱处理Colo-16细胞株24h后,用流式细胞仪测定不同浓度氧化苦参碱组Colo-16细胞株凋亡率,用免疫印迹(western blot)法测定不同浓度组细胞自噬相关蛋白LC3-Ⅰ、LC3-Ⅱ的表达及LC3-Ⅰ→LC3-Ⅱ转化率.同时用western blot法检测哺乳动物雷帕霉素靶蛋白(mTOR)、mTOR调节相关蛋白(Raptor)及其mTORser2448、ser2481位点磷酸化产物和Raptor ser792位点磷酸化产物的表达.结果:1.000 g/L、2.000 g/L、4.000 g/L氧化苦参碱浓度组凋亡率明显高于阴性对照组,差异有统计学意义(F=355.12,P＜0.05),不同浓度组之间凋亡率差异也有统计学意义(P＜0.05).western blot法发现,阴性对照组、溶剂对照组及氧化苦参碱分别为0.025 g/L、0.050 g/L、0.100 g/L、0.500 g/L、1.000 g/L、2.000 g/L各组之间LC3-Ⅰ、LC3-Ⅱ的表达及LC3-Ⅰ→LC3-Ⅱ转化率比较,差异无统计学意义(F值分别为0.86,0.63,0.33,P＞0.05),各组别之间mTOR、Raptor蛋白及其mTORser2448、ser2481位点磷酸化产物和Raptor ser792位点磷酸化产物的表达,差异均无统计学意义(F值分别为1.33、1.42、1.12、0.95及0.47,P＞0.05).结论:氧化苦参碱可上调Colo-16细胞株的凋亡,且与其浓度正相关.氧化苦参碱对细胞自噬的水平无影响,对自噬相关调控蛋白的表达也无影响.因此,氧化苦参碱可能通过上调Colo-16细胞株的凋亡而非自噬来实现其抗肿瘤效应.“,”Objective:To evaluate effects of oxymatrine on apoptosis and autophagy of Colo-16 cells,a cutaneous squamous cell carcinoma cell line.Methods:The Colo-16 cells were treated with oxymatrine at different concentrations(0.025 g/L,0.050g/L,0.100 g/L,0.500 g/L,1.000 g/L,2.000 g/L,and 4.000 g/L group)for 24 h.Flow cytometry was performed to evaluate the apoptosis rates of Colo-16 cells.Western blot was used to assess expression levels of LC3-Ⅰ,LC3-Ⅱ,mTOR,Raptor,phos-phor-mTOR (Ser2448),phosphor-mTOR (Ser2481),phosphor-Raptor (Ser792),and the inversion rate of LC3-Ⅰ to LC3-Ⅱ.Results:In comparison to negative control,apoptosis rates increased significantly following the treatment with 1 g/L,2 g/L or 4 g/L of oxymatrine(F=355.12,P＜0.05).And apoptosis rates varied significantly among groups treated with these three concentrations(P＜0.05).The expression levels of LC3-Ⅰ,LC3-Ⅱ and the inversion rate of LC3-Ⅰ to LC3-Ⅱ were no significant differences among negative control,solvent-treated and groups treated with oxymatrine at concentrations of 0.025 g/L,0.05 g/L,0.10g/L,0.5 g/L,1 g/L,or 2g/L(F=0.86,0.63,0.33,all P＞0.05).Similarly,expression levels of mTOR,Raptor,phos1phor-mTOR(Ser2448),phosphor-mTOR(Ser2481),phosphor-Raptor(Ser792) were no significant differences among these groups(F=1.33,1.42,1.12,0.95,0.47,all P＞0.05).Conclusions:Oxymatrine can up-regulate the apoptosis of Colo-16 cells at dose-dependent manner.Oxymatrine has no effect on autophagy of Colo-16 cells and autophagy associated proteins.Antitumor benefit of oxymatrine could be via up-regulating apoptosis,but not autophagy.