过表达Beclin1可促进黑色素瘤A375细胞的自噬并抑制其上皮间质转化  

Overexpression of Beclin1 enhances autophagy and inhibits epithelial mesenchymal transition in melanoma A375 cells

作　　者：韩昭[1] 高艳玲 张建忠[1] 张伶[1] 徐艳艳 李小静[1] HAN Zhao;GAO Yanling;ZHANG Jianzhong;ZHANG Ling;XU Yanyan;LI Xiaojing(Dermatology Department,Affiliated Hospital of Hebei Engineering University;Emergency Department,Handan Central Hospital Emergency,Handan 056002,China)

机构地区：[1]河北工程大学附属医院皮肤科 [2]邯郸市中心医院急诊科,河北邯郸056002

出　　处：《皮肤性病诊疗学杂志》2022年第3期206-211,共6页Journal of Diagnosis and Therapy on Dermato-venereology

基　　金：邯郸市科学技术研究与发展计划项目(19422083034ZC)。

摘　　要：目的探讨Beclin1的过表达对黑色素瘤A375细胞自噬及EMT过程的影响。方法采用脂质体转染法将人恶性黑色素瘤细胞A375分为3组,分别为空白组(只转染试剂,无质粒)、NC组(转染空质粒)、转染组(转染携带Beclin1质粒),3组细胞分别接种于3组裸鼠皮下,观察成瘤后21 d各组移植瘤的生长情况。采用RT-PCR和Western blotting分别检测各组肿瘤组织中Beclin1、LC3Ⅱ、Vimentin、N-cadherin和E-cadheirn mRNA及蛋白表达水平。结果3组接种A375细胞的裸鼠成瘤率100%,转染组肿瘤生长速度低于空白组和NC组,差异具有统计学意义(P<0.05);RT-PCR结果显示,空白组与NC组比较,各基因mRNA的表达无明显差异(P>0.05);相对于空白组与NC组,转染组Beclin1、LC3Ⅱ和E-cadherin的mRNA表达均升高,Vimentin和N-cadherin的mRNA表达均降低,差异具有统计学意义(均P<0.05)。Western blotting结果显示,空白组与NC组比较,各基因蛋白的表达无明显差异(P>0.05);相对于空白组与NC组,转染组Beclin1、LC3Ⅱ和E-cadherin蛋白表达均明显升高,Vimentin和N-cadherin蛋白表达均明显降低,差异均具有统计学意义(均P<0.05)。结论Beclin1的过表达可促进黑色素瘤细胞发生自噬,并抑制其EMT过程。Objective To investigate the effect of Beclin1 overexpression on autophagy and EMT in melanoma A375 cells.Methods Human malignant melanoma A375 cells were divided into three groups by liposome transfection,i.e.,blank group(no plasmid),NC group(transfection empty plasmid),and transfection group(transfection with Beclin1).These three groups were inoculated subcutaneously in three groups of nude mice for 21 days after tumor formation.The expression levels of mRNA and proteins,including Beclin1,LC3Ⅱ,Vimentin,N-cadherin and E-cadheirn,were detected by RT-PCR and Western blotting,respectively.Results Although tumor developed in all three groups,the tumor growth rate was significantly lower in the transfection group than in either blank or NC groups(P<0.05).Expression levels of both mRNA and proteins were similar between blank group and the NC group(P>0.05).Compared with the blank and NC groups,the transfection group displayed significantly higher expression levels of both mRNA and proteins for Beclin1,LC3Ⅱand E-cadherin(P<0.05),and lower expression levels of mRNA and proteins for Vimentin and N-cadherin(P<0.05).Conclusion Beclin1 overexpression enhances autophagy and inhibits EMT in melanoma cells.

关 键 词：BECLIN1 过表达 自噬 黑色素瘤 上皮间质转化 

分 类 号：R73[医药卫生—肿瘤]