赖氨藤黄酸对乳腺癌MCF-7细胞增殖和凋亡的影响及其作用机制  

Influence of gambogic acid lysinate in proliferation and apoptosis in breast cancer MCF-7 cells and its mechanism

作　　者：魏洁[1] 李开济[2] 魏静波[2] 赵毓芳[2] 闫丰[3] 吕翠平[1] 甄永占[2] 

机构地区：[1]卫生部北京医院卫生部北京老年医学研究所,北京100730 [2]河北联合大学基础医学院组织学与胚胎学教研室,河北唐山063000 [3]河北联合大学附属医院肿瘤科,河北唐山063000

出　　处：《吉林大学学报（医学版）》2015年第3期476-480,I0004,共6页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(81001439);河北省科技厅自然科学基金资助课题(H2012401030);河北联合大学大学生创新性实验计划资助课题(X2014078)

摘　　要：目的:研究赖氨藤黄酸(GAL)对乳腺癌MCF-7细胞增殖和凋亡的影响并探讨其作用机制,为GAL抗乳腺癌的临床研究和应用提供理论依据。方法:选取处于对数生长期的MCF-7细胞,并随机分为空白对照组、不同剂量(0.25、0.50、1.00、2.00、4.00和8.00μmol·L-1)GAL组和不同剂量(0.25、0.50、1.00、2.00、4.00和8.00μmol·L-1)藤黄酸组,采用MTT法检测各组MCF-7细胞48h增殖活性。选取对数生长期的MCF-7细胞,2μmol·L-1 GAL作用MCF-7细胞不同时间(0、6、12、24、36、48和60h),采用MTT法检测作用不同时间后各组MCF-7细胞增殖活性。流式细胞术和Hoechst 33258染色检测各组MCF-7细胞24h时凋亡情况,采用Western blotting法检测各组MCF-7细胞24h时凋亡相关蛋白的表达水平。结果:细胞培养24h后,不同剂量GAL组MCF-7细胞存活率低于空白对照组(P<0.05),随剂量增加和作用时间延长细胞存活率下降(P<0.05)。流式细胞术和Hoechst 33258染色,与空白对照组比较,不同剂量GAL组MCF-7细胞凋亡率明显升高(P<0.05),且呈剂量依赖性。Western blotting检测,与空白对照组比较,不同剂量GAL组细胞沉默信息调节因子1(SIRT1)蛋白表达水平明显降低(P<0.05),而caspase-3切割片段蛋白的表达水平明显高于对照组(P<0.05)。结论:GAL通过抑制SIRT1蛋白表达水平和上调caspase-3切割片段蛋白的表达水平诱导乳腺癌MCF-7细胞凋亡。Objective To investigate the effect of gambogic acid lysinate（GAL）on the proliferation and apoptosis of human breast cancer MCF-7cells and its mechanism,and to provide theoretical foundation for clinical trial and use of GAL against breast cancer.Methods The human breast cancer MCF-7cells in logarithm growth phase were selected and randomly divided into blank control group,different doses（0.25,0.50,1.00,2.00,4.00,8.00μmol·L^-1）of GAL groups,and different doses（0.25,0.50,1.00,2.00,4.00,8.00μmol·L^-1）of gambogic acid groups.After the MCF-7cells were treated with GAL or gambogic acid for 48 h,MTT assay was used to detect the proliferation activities of MCF-7cells.After the MCF-7cells in logarithm growth phase were treated with 2μmol·L^-1 GAL for different time（0,6,12,24,36,48,60h）,MTT assay was also used to detect the proliferation activities of MCF-7cells in various groups.The flow cytometry method and Hoechst 33258 staining were used to analyze the apoptosis of MCF-7cells in various groups.The expression levels of apoptosisassociated proteins were detected by Western blotting method.Results After 48 hcell culture,the cell viabilities in GAL groups were lower than those in control group（P〈0.05）and the cell viability was inhibited in a dosedependent manner（P〈0.05）.After the MCF-7cells were treated by 2μmol·L^-1 GAL for different time,the cell proliferation was inhibited in a time-dependent manner（P〈0.05）.The flow cytometry method and Hoechst33258 staining results showed that the apoptotic rates of MCF-7cells in GAL groups were higher than those in control group（P〈0.05）.The expressions levels of SIRT1 in GAL groups were significantly lower than that in control group（P〈0.05）,while the expression amounts of cleaved caspase-3were higher than that in control group（P〈0.05）.Conclusion GAL can induce the apoptosis of breast cancer MCF-7cells by inhibiting the expression level of SIRT1 and up-regulating the expression amount of cleaved caspase-3.

关 键 词：赖氨藤黄酸 乳腺肿瘤 细胞凋亡 沉默信息调节因子1 

分 类 号：R737.9[医药卫生—肿瘤]