氧化苦参碱对皮肤鳞状细胞癌Colo-16细胞株凋亡及自噬影响的研究  

Effects of oxymatrine on the apoptosis and autophagy of Colo-16 cells

作　　者：张青松[1] 陈旭[2] 黄丹[2] 顾恒[2] 

机构地区：[1]常熟市中医院皮肤科,江苏常熟215500 [2]中国医学科学院北京协和医学院皮肤病研究所理疗科,江苏南京210042

出　　处：《临床皮肤科杂志》2017年第12期830-834,共5页Journal of Clinical Dermatology

基　　金：苏州市科技局立项课题(SYSD2013017);常熟市科技局立项资助课题(CS201319)资助项目

摘　　要：目的:探讨氧化苦参碱对皮肤鳞状细胞癌(c SCC)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24 h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(m TOR)、m TOR调节相关蛋白(Raptor)及其m TORser2448、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m TOR、Raptor蛋白及其m TORser2448、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, 0.050 g/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 con- centrations（P〈0.05）. The expression levels of LC3-I, LC3-II and the inversion rate of LC3-I to LC3-II were no signifi- cant differences among negative control, solvent-treated and groups treated with oxymatrine at concentrations of 0.025 g/L, 0.05 g/L, 0.10 g/L, 0.5 g/L, 1 g/L, or 2 gL（F=0.86,0.63,0.33, all /9〉0.05）. Similarly, expression levels of mTOR,Raptor, phos- phor-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. Anti- tumor benefit of oxymatrine could be via up-regulating apoptosis, but not autophagy.

关 键 词：氧化苦参碱 皮肤鳞状细胞癌 凋亡 自噬 

分 类 号：R739.5[医药卫生—肿瘤]