p-AMPK介导的不同亚型乳腺癌细胞自噬及其分子机制  

Autophagy in different subtypes of breast cancer cells mediated by p-AMPK and its molecular mechanisms

作　　者：杨鑫娇 胡汝瑶 熊喆 邹迪 蔡洁[1] 夏从龙 白忠彬[1] 赵红业[1] YANG Xin-jiao;HU Ru-yao;XIONG Zhe;ZOU Di;CAI Jie;XIA Cong-long;BAI Zhong-bin;ZHAO Hong-ye(Key Laboratory of Gene Editing and Xenotransplantation for Miniature Pigs in Yunnan Province,Yunnan Agricultural University,Kunming 650201,China;College of Pharmacy,Dali University,Dali Yunnan 671000,China)

机构地区：[1]云南农业大学云南省小型猪基因编辑与异种器官移植重点实验室,云南昆明650201 [2]大理大学药学院,云南大理671000

出　　处：《中国药理学通报》2025年第5期898-907,共10页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81872452)。

摘　　要：目的探究p-AMPK活性对不同亚型MDA-MB-231(三阴性乳腺癌细胞)和MCF-7(雌激素受体阳性细胞)发生自噬的影响及调节机制。方法用EBSS、Baf-A1和EBSS+Baf-A1处理MDA-MB-231细胞4 h,MCF-7细胞8 h后,自噬对细胞的增殖及凋亡影响,观察细胞线粒体,检测自噬标志物LC3B、P62、LAMP1、TOM20、AMPK、p-AMPK、ULK1和Beclin1/VPS34蛋白表达情况,抑制AMPK活性验证自噬通路。结果饥饿诱导后乳腺癌发生自噬,不同亚型乳腺癌细胞自噬进程不一致;抑制细胞增殖;MDA-MB-231细胞自噬发生后p-AMPK水平上升,ULK1水平降低,通过p-AMPK激活ULK1启动自噬;MCF-7细胞自噬发生后p-AMPK和ULK1水平均降低,说明其激活自噬可能不由p-AMPK介导。结论MDA-MB-231主要通过p-AMPK直接激活ULK1的方式启动自噬,不依赖MTOR途径;MCF-7细胞可能通过AMPK活性抑制激活MTOR或其他上游因子直接激活MTOR的方式启动自噬,根据不同亚型细胞自噬通路调节p-AMPK活性,能够更精准靶向治疗不同类型的乳腺癌。Aim To investigate the effect of p-AMPK activity on autophagy in different subtypes of MDA-MB-231(triple-negative breast cancer cells)and MCF-7(estrogen receptor-positive cells)and its regulatory mechanism.Methods MDA-MB-231 cells were treated with EBSS,Baf-A1,and EBSS+Baf-A1 for four hours,and MCF-7 cells for eight hours.The effects of autophagy on cell proliferation and apoptosis were observed,mitochondrial morphology was examined,and the expression of autophagy markers LC3B,P62,LAMP1,TOM20,AMPK,p-AMPK,ULK1,and Beclin1/VPS34 proteins was detected.The autophagy pathway was validated by inhibiting AMPK activity.Results Breast cancer cells underwent autophagy after starvation induction(EBSS),with inconsistent autophagy processes observed in different subtypes of breast cancer cells.Autophagy inhibited cell proliferation.In MDA-MB-231 cells,autophagy led to an increase in p-AMPK levels and a decrease in ULK1 levels,initiating autophagy through p-AMPK activation of ULK1.In MCF-7 cells,both p-AMPK and ULK1 levels decreased after autophagy,suggesting that autophagy might not be mediated by p-AMPK activation.Conclusions MDA-MB-231 cells primarily initiate autophagy by directly activating ULK1 by p-AMPK,independent of the MTOR pathway.In MCF-7 cells autophagy might be triggered by inhibiting MTOR through AMPK activity or directly activating MTOR through other upstream factors.Regulating p-AMPK activity based on the autophagy pathways in different cell subtypes could enable more precise targeting and treatment of different types of breast cancer.

关 键 词：p-AMPK 乳腺癌 饥饿诱导 雌激素受体 自噬 MTOR 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R392[医药卫生—基础医学] R392.11R737.9