MAPK4表达对黑色素瘤细胞增殖和凋亡的影响  

Effect of MAPK4 expression on proliferation and apoptosis of melanoma cells

作　　者：赵晟辰 邓朝伟 刘乐凡 郑琪 张珂浛 赵凌宇 吴锋[4] ZHAO Shengchen;DENG Chaowei;LIU Lefan;ZHENG Qi;ZHANG Kehan;ZHAO Lingyu;WU Feng(First Clinical Medi-cal College of Xi'an Jiaotong University Health Science Center,Xi'an 710061,China;Department of Cell Biology and Genetics,School of Basic Medical Sciences,Xi'an Jiaotong University Health Science Center;Department of Medical Oncology,Shaanxi Provin-cial Tumor Hospital;Center of Teaching and Experiment for Medical Post Graduates,School of Medicine,Xi'an Jiaotong University)

机构地区：[1]西安交通大学医学部第一临床医学院,西安710061 [2]西安交通大学医学部基础医学院细胞生物学与遗传学系 [3]陕西省肿瘤医院肿瘤内科 [4]西安交通大学医学部研究生教学实验中心

出　　处：《山西医科大学学报》2025年第1期8-14,共7页Journal of Shanxi Medical University

基　　金：陕西省重点研发计划项目(2023-YBSF-178);国家级大学生创新训练计划项目(202310698152)。

摘　　要：目的 探讨丝裂原活化蛋白激酶4(MAPK4)在黑色素瘤中的表达及影响细胞增殖、细胞周期和凋亡的分子机制。方法 采用qRT-PCR和免疫组织化学法检测人黑色素瘤组织和毗邻癌旁组织中MAPK4表达变化。A2058细胞分为阴性对照组、siRNA-1组、siRNA-2组,分别转染NC-siRNA、MAPK4 siRNA1和MAPK4 siRNA2。于转染12,36,60,84 h通过MTT法观察各组细胞增殖活力的变化,采用细胞克隆实验检测细胞增殖。应用流式细胞技术分析细胞周期和细胞凋亡的变化。应用Western blotting检测各组MAPK4、AKT的蛋白表达。结果 与毗邻癌旁组织比较,黑色素瘤组织中MAPK4 mRNA和蛋白表达均显著增加(P<0.01)。与NC-siRNA组相比,MAPK4 siRNA-1组和MAPK4 siRNA-2组A2058细胞增殖明显降低(P<0.01),G0/G1期细胞显著增加(P<0.01),S期细胞显著减少(P<0.01),凋亡细胞数量显著增加(P<0.01),MAPK4和AKT蛋白的表达显著下调(P<0.01)。结论 MAPK4在黑色素瘤组织中表达上调,沉默MAPK4可能通过调节AKT信号通路抑制黑色素瘤细胞增殖、周期转换,促进细胞凋亡。Objective To explore the expression of mitogen-activated protein kinase 4(MAPK4)in melanoma and its effect and mo-lecular mechanisms on cell proliferation,cell cycle and apoptosis.Methods qRT-PCR and immunohistochemistry were employed to detect MAPK4 expression in adjacent non-tumor tissues and melanoma tissues.A2058 cells were divided into negative control group,siRNA-1 group and siRNA-2 group,and transfected with NC-siRNA,MAPK4 siRNA1 and MAPK4 siRNA2,respectively.The cell proliferation activity was assessed using MTT method at 12,36,60,84 h after transfection,and the cell proliferation was detected by cell cloning assay.Cell cycle and apoptosis were analyzed by flow cytometry.The expressions of MAPK4 and AKT proteins were de-tected by Western blotting.Results Both MAPK4 mRNA and protein expressions were significantly increased in melanoma tissues compared to adjacent non-tumor tissues(P<0.01).Compared with NC-siRNA group,the cell proliferation was significantly reduced in MAPK4 siRNA-1 group and MAPK4 siRNA-2 group(P<0.01),the percentage of G0/G1 phase cells was significantly increased,the percentage of S phase cells was decreased(P<0.01),the early and late apoptotic rates were increased(P<0.01),and MAPK4 and AKT protein expressions were downregulated(P<0.01).Conclusion MAPK4 expression is upregulated in melanoma tissues,and silencing MAPK4 may inhibit melanoma cell proliferation and cell cycle transition,and promote apoptosis through modulating AKT signaling pathway.

关 键 词：MAPK4 黑色素瘤 细胞增殖 细胞周期 细胞凋亡 AKT信号通路 

分 类 号：R739.5[医药卫生—肿瘤]