CDC25C表达下调对黑色素瘤细胞B16自噬和迁移能力的影响  

Effects of down-regulation of CDC25C expression on autophagy and migration ability of melanoma B16 cell line

作　　者：郑方园 潘桂珍 钟婷 李婷玉 陈宇露 李京原 陈琪琦 陈红 王桂闽 陆小楠 莫发荣[1] Zheng Fangyuan;Pan Guizhen;Zhong Ting;Li Tingyu;Chen Yulu;Li Jingyuan;Cheng Qiqi;Chen Hong;Wang Guimin;Lu Xiaonan;Mo Farong(School of Basic Medical Sciences,Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学基础医学院,南宁530021

出　　处：《广西医科大学学报》2022年第2期221-226,共6页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81160264);广西自然科学基金资助项目(No.2018GXNSFAA281071);广西医科大学大学生创新创业计划资助项目(No.201910598106,No.201910598252)。

摘　　要：目的:利用慢病毒质粒构建细胞分裂周期蛋白25(CDC25)表达下调的黑色素瘤B16细胞,探讨CDC25C表达下调对黑色素瘤细胞自噬和迁移能力的影响。方法:利用TCGA可视化在线工具GEPIA数据库检测CDC25C在黑色素瘤中的表达;通过慢病毒质粒转染构建CDC25C表达下调的黑色素瘤B16细胞株,并以RT-qPCR技术确认其表达量;以RT-qPCR和Western blotting方法检测CDC25C表达下调对自噬相关分子p62、LC3及beclin1的影响;划痕修复实验观察CDC25C表达下调对B16细胞迁移能力的影响。结果:生物信息学分析发现CDC25C在黑色素瘤中高表达(P<0.05);成功构建CDC25C表达下调的B16细胞株;与对照组相比,CDC25C表达下调的B16细胞中自噬相关分子p62、LC3、beclin1 mRNA和蛋白表达量均增高(均P<0.05),细胞增殖迁移率降低(P<0.001)。结论:CDC25C表达下调能激活黑色素瘤细胞自噬过程,并抑制细胞的迁移能力。Objective:To construct melanoma B16 cell line with down-regulated CDC25C expression by using lentiviral plasmids,and to investigate the effects of down-regulated CDC25C expression on autophagy and migration ability of melanoma cells.Methods:CDC25C expression in B16 cells was detected by Gene Expression Profiling Interactive Analysis(GEPIA)database,a visual online tool of the Cancer Genome Atlas(TCGA).The melanoma B16 cell line with down-regulated CDC25C expression was constructed by lentiviral plasmid transfection,and its expression level was confirmed by RT-qPCR method.The effects of down-regulation of CDC25C expression on autophagy-related molecules p62,LC3 and beclin1 were detected by RT-qPCR and western blotting method.the effects of down-regulation of CDC25C on the migration ability of B16 cells were evaluated by Wound healing assay.Results:Bioinformatics analysis showed that CDC25C was highly expressed in melanoma(P<0.05).A stable B16 cell line with down-regulated CDC25C was successfully constructed.Compared with the control group,mRNA and protein expression levels of autophagy-related molecules p62,LC3,and beclin1 in B16 cells with down-regulated CDC25C were increased(P<0.05),and the cell migration ability was obviously decreased(P<0.001).Conclusion:Downregulation of CDC25C expression can activate melanoma cell autopha-gy,and inhibit its cell migration ability.

关 键 词：CDC25C 黑色素瘤 自噬 迁移 

分 类 号：R739.5[医药卫生—肿瘤]