3-甲基腺苷对芹菜素诱导乳腺癌T47D细胞系自噬和凋亡的影响  

Effect of 3-Methyladenine on Apigenin-Induced Autophagy and Apoptosis in the Breast Cancer T47D Cell Line

作　　者：刘博文[1] 张斌[1] 张伟然[1] 张月[1] 冯炜红[1] 李媛媛[1] 曹旭晨[1] 

机构地区：[1]天津医科大学附属肿瘤医院乳腺一科乳腺癌防治教育部重点实验室天津市肿瘤防治重点实验室,天津市300060

出　　处：《中国肿瘤临床》2011年第21期1318-1321,共4页Chinese Journal of Clinical Oncology

基　　金：国家自然科学基金(编号:81001186);天津市自然科学基金(编号:10JCYBJC14100)资助~~

摘　　要：目的:研究自噬抑制剂3-甲基腺苷(3-methylaclenine,3-MA)对芹菜素诱导乳腺癌T47D细胞系自噬和凋亡的影响。方法:常规培养人乳腺癌T47D细胞,分为对照组、3-MA组、芹菜素组、3-MA+芹菜素组。MTT法检测各组的细胞增殖抑制率;GFP-LC3质粒转染观察各组细胞自噬情况;Hochest/Mito-Red/YO-PRO-1染色法观察各组细胞凋亡形态;Annexinv/PI双染法流式细胞仪检测各组细胞凋亡率;western hlot法检测LC3及PARP的变化。结果:MTT示3-MA+芹菜素组的增殖抑制率明显高于其他各组(P<0.05);GFP-LC3质粒转染结果显示:对照组,3-MA组及3-MA+芹菜素组自噬不明显,而芹菜素组与其他各组相比自噬明显增多(P<0.05);3-MA+芹菜素组的凋亡细胞增多,各组凋亡率分别为(12.73±0.05)%,(18.46±0.03)%,(23.27±0.07)%,(34.14±0.05)%,与对照组相比有统计学意义(P<0.05);Western blot结果显示,芹菜素组LC3-Ⅱ增高,而3-MA组及3-MA+芹菜素组的LC3-Ⅱ显著减少,3-MA+芹菜素组PARP的剪切带相比其他各组明显增加。结论:自噬抑制剂3-MA抑制细胞自噬后,能够明显增强芹菜素对乳腺癌T47D细胞系的凋亡诱导效应。Objective： To study the effect of 3-methyladenine （ 3-MA ） on apigenin-induced autophagy and apoptosis in the breast cancer T47D cell line. Methods： T47D cells cultured in vitro were divided into control, 3-MA, apigenin, and 3-MA＋apigenin groups. MTT assay was used to detect the inhibition rate of each group. GFP-LC3 plasmid transfection was used to observe the autopha- gy of each group by fluorescence microscopy. Hoechst/Mito-Red/YO-PRO-1 staining was used to observe the cell apoptosis of each group using fluorescence microscopy. AnnexinV/PI staining was used to detect the apoptotic rate by flow cytometry. Western blot analy- sis was performed to detect changes in LC3 and PARP. Results： MTT assay showed that the inhibitory rate of the 3-MA＋apigenin group was significantly higher than those of the other groups （ P 〈 0.05 ）. GFP-LC3 plasmid transfection showed that autophagy was not obvious in the control, 3-MA, and 3-MA＋apigenin groups but was significantly increased in the apigenin group （ P 〈 0.05 ）. Apoptosis increased in the 3-MA＋apigenin group, and the apoptotic rates of the four groups were （ 12.73 ± 0.05 ） %, （ 18.46 ±0.03 ） %, （23.27 ±0.07 ） %, and （ 34.14 ±0.05 ） %, respectively （ P 〈 0.05 ）. Western blot analysis showed that LC3-II levels increased in the apigenin group but significantly decreased in the 3-MA and 3-MA＋apigenin groups. Cleaved PARP significantly increased in the 3-MA＋apigenin group compared with the other groups. Conclusion： The inhibition of autophagy by 3-MA could significantly increase apigenin-induced apoptosis in the breast cancer T47D cell line.

关 键 词：芹菜素 3-甲基腺苷 乳腺癌细胞 凋亡 

分 类 号：R737.9[医药卫生—肿瘤]