[关键词]
[摘要]
目的 探讨龙葵碱对人胶质瘤U251细胞增殖及凋亡的影响。方法 使用不同质量浓度的龙葵碱处理U251细胞,采用CCK-8法、划痕法、Transwell法、Hochest法、流式细胞术、免疫印迹法分别检测细胞增殖率、细胞迁移与侵袭、细胞凋亡及凋亡相关蛋白的表达。结果 龙葵碱对U251细胞的48 h半数抑制浓度(IC50)为20.05 μg/μL,其质量浓度在5~35 μg/μL时,能够抑制细胞增殖(P< 0.05);与对照组相比,龙葵碱组抑制细胞迁移和侵袭的能力随药物质量浓度增高而增强(P< 0.05),同时细胞也出现明显凋亡特征,并均呈浓度依赖性;免疫印迹法显示龙葵碱可使凋亡相关蛋白Bax表达增加,Bcl-2表达降低,Bcl-2/Bax值降低。结论 龙葵碱在有效剂量(5~35 μg/μL)对U251细胞增殖生长具有抑制作用,并呈浓度依赖性;龙葵碱可抑制U251细胞的侵袭与迁移,可以影响凋亡相关蛋白Bax和Bcl-2的表达,使Bcl-2/Bax值降低,从而抑制U251细胞的增殖生长。
[Key word]
[Abstract]
Objective To investigate the effect of solanine on growth and proliferation of U251 cells. Methods U251 cells were cultured with different concentration of solanine together. CCK-8 method, wound healing assay, Transwell method, Hochest assay, flow cytometry screening and Western blotting were used to detect inhibitory rate, migration and invasive rates, apoptosis rate and expression level of cell apoptosis-related proteins. Results CCK-8 assay showed the median inhibition concentration (IC50) of 48 h was 20.05 μg/μL. It was effective that solanine in the concentration range of 5—35 μg/μL could inhibit the growth and proliferation of U251 cells, and played a role in a dose-dependent manner (P <0.05). Compared with control group, experimental groups can not only inhibit U251 cell migration and invasion in a dose-dependent manner (P <0.05), but also appear some significant apoptosis characteristics with a concentration dependence. Western blotting assayed that the expression of Bax protein was upregulated, whereas Bcl-2 protein expression was downregulated in experimental groups compared with control group and these changes were dose-dependent. Conclusion Solanine can inhibit growth and proliferation of U251 cells in effective concentration and in a dose-dependent manner. Solanine can inhibit the invasion and migration of U251 cells. Solanine can induce the apoptosis of U251 cells, and inhibit the proliferation and growth of U251 cells via regulating the expression of Bax and Bcl-2 proteins and affecting Bcl-2/Bax ratio.
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[基金项目]
国家自然科学基金资助项目(81502161);重庆市科委基础与前沿项目(cstc2014jcyjA10028);重庆医科大学基础医学院项目(2013-5)