α-苦瓜素通过激活JNK信号通路诱导口腔鳞癌细胞凋亡的机制研究  

Mechanism ofα-MMC in Oral Squamous Cell Carcinoma HSC-3 cells based on JNK signaling pathway

作　　者：冷平[1] 蒋佩文 杜文倩 蔡紫微 张清亮 汪林 吴泳洁 黄敏 李敏惠 LENG Ping;JIANG Pei-wen;DU Wen-qian;CAI Zi-wei;ZHANG Qin-liang;WANG Lin;WU Yong-jie;HUANG Min;LI Min-hui(College of Medical Technology,Chengdu University of Traditional Chinese Medicine,Chengdu 610075;Department of Laboratory Medicine,Jintang First People's Hospital,Chengdu 610400;School of Basic Medicine,Chengdu Medical College,Chengdu 610500,Sichuan,China;School of Pharmacy,Chengdu Medical College,Chengdu 610500,Sichuan,China;School of Bioscience and Technology,Chengdu Medical College,Chengdu 610500,Sichuan,China;Center of Scientific Research and Experiment,Chengdu Medical College,Chengdu 610500,Sichuan,China)

机构地区：[1]成都中医药大学医学技术学院,四川成都610075 [2]金堂县第一人民医院实验医学科,四川成都610400 [3]成都医学院基础医学院,四川成都610500 [4]成都医学院药学院,四川成都610500 [5]成都医学院生物科学与技术学院,四川成都610500 [6]成都医学院科研实验中心,四川成都610500

出　　处：《川北医学院学报》2022年第9期1101-1105,共5页Journal of North Sichuan Medical College

基　　金：四川省科技厅项目(2022NSFSC0690,2020YFS0321);四川省大学生创新创业项目(202113705009);四川省发育与再生重点实验室开放项目(SYS20-06);成都医学院研究生科研创新项目(YCX2020-16);成都医学院四川省养老保健协同创新中心项目(YLZBZ2007)。

摘　　要：目的:探讨α-苦瓜素(α-MMC)对口腔鳞状细胞癌HSC-3细胞的增殖抑制作用及通过激活细胞信号传导通路调控细胞凋亡的分子机制。方法:通过CCK-8法检测α-MMC对HSC-3细胞的增殖抑制作用;利用PI、Annexin V-FITC/PI、JC-1染色及流式细胞术测定α-MMC对HSC-3细胞周期、凋亡、线粒体膜电位的影响;通过蛋白免疫印迹技术判断α-MMC作用后,HSC-3细胞凋亡相关蛋白及信号通路蛋白的表达情况。结果:CCK-8分析结果显示,α-MMC以药物浓度和时间梯度依赖的方式抑制HSC-3细胞增殖,其24 h和48 h的IC50值分别为50.38μg/mL和20.17μg/mL;流式细胞术检测发现,α-MMC阻滞HSC-3细胞周期于G2/M期,降低细胞线粒体膜电位,诱导细胞凋亡;蛋白免疫印迹结果显示,α-MMC上调促凋亡蛋白Bim表达,下调抗凋亡蛋白Mcl-1的表达;激活JNK信号通路,即上调p-MKK4、p-JNK、p-c-Jun的表达水平。结论:α-MMC对口腔鳞状细胞癌HSC-3细胞具有增殖抑制作用,其机制可能是α-MMC激活JNK信号通路触发细胞周期阻滞,并通过线粒体途径诱导细胞凋亡。Objective:To explore the inhibitory effect ofα-MMC on the proliferation of Oral Squamous Cell Carcinoma HSC-3 cells and explain its molecular mechanism of regulating apoptosis by activating cell signaling pathways.Methods:CCK-8 method was used to explore the inhibitory effect ofα-MMC on the proliferation of HSC-3 cells.PI,Annexin V-FITC/PI,JC-1 fluorescence dye and flow cytometry were used to detect the effects ofα-MMC on the cell cycle proportion,mitochondrial membrane potential and apoptosis of HSC-3 cells.Western blot was used to detect the expression of apoptosis-related and signal pathway-related proteins in HSC-3 cells treated withα-MMC.Results:CCK-8 analysis showed thatα-MMC inhibited the proliferation of HSC-3 cells in a concentration-dependent and time-dependent manner,and its IC50 values at 24 h and 48 h were 50.38μg/mL and 20.17μg/mL,respectively.The results from flow cytometry showed thatα-MMC increased the proportion of G2/M,decreased the mitochondrial membrane potential,and induced apoptosis in HSC-3 cells.Western blot showed thatα-MMC could down-regulate the expression of Mcl-1,up-regulate the expression of Bim,p-MKK4,p-JNK and p-c-Jun.Conclusion:α-MMC can inhibit the proliferation of Oral Squamous Cell Carcinoma HSC-3 cells,and the possible mechanism is thatα-MMC activates JNK signaling pathway to induce apoptosis of HSC-3 cells by mitochondrial pathway and G2/M arrest.

关 键 词：α-苦瓜素 口腔鳞癌 JNK信号通路 线粒体途径 

分 类 号：R575[医药卫生—消化系统]