microRNA-136基于Wnt信号通路在黑素瘤EMT中的作用  

Role of MicroRNA-136 in EMT of Melanoma Based on Wnt Signaling Pathway

作　　者：韩昭[1] 王久江[1] 张伶[1] 张建忠[1] 牛亮 刘钊 徐艳艳 刘志军[1] 李小静[1] HAN Zhao;WANG Jiujiang;ZHANG Ling;ZHANG Jianzhong;NIU Liang;LIU Zhao;XU Yanyan;LIU Zhijun;LI Xiaojing(Department of Dermatology,Affiliated Hospital of Hebei University of Engineering,Handan 056000,China)

机构地区：[1]河北工程大学附属医院皮肤科,河北邯郸056000

出　　处：《中国皮肤性病学杂志》2020年第2期128-133,共6页The Chinese Journal of Dermatovenereology

摘　　要：目的探讨microRNA-136(miR-136)基于Wnt信号通路在黑素瘤上皮-间充质细胞转化(EMT)中的调控作用。方法常规培养正常小鼠A9细胞(阴性对照组)和小鼠黑素瘤B16细胞,采用脂质体转染法将B16细胞分为空白组(只转染试剂,无转染序列)、NC组(转染miR-136 nc)、miR-136模拟组(转染miR-136模拟物)、miR-136抑制组(转染miR-136抑制剂)。通过qRT-PCR法检测各组细胞中miR-136的表达,qRT-PCR及Western blot实验分别检测各组细胞Wnt3a、β-catenin、Ecadherin及N-cadherin mRNA及蛋白表达的差距。结果与阴性对照组相比,其他各组miR-136表达均显著降低,空白组与NC组比较差异无统计学意义(P>0.05)。相对于空白组与NC组,miR-136模拟组miR-136表达显著增加(P<0.05);miR-136抑制组miR-136的表达明显降低(P<0.05)。与对照组相比,其他各组E-cadherin mRNA及蛋白表达显著降低(P均<0.05)。β-catenin、Wnt3a、N-cadherin的mRNA及蛋白表达显著增加(P均<0.05)。空白组与NC组比较差异无统计学意义(P>0.05)。与空白组和NC组相比,miR-136模拟组E-cadherin的mRNA及蛋白表达显著增加(P<0.05);β-catenin、Wnt3a和N-cadherin的mRNA及蛋白表达均显著降低(P均<0.05)。miR-136抑制组E-cadherin的mRNA及蛋白表达显著降低(P均<0.05),而β-catenin、wnt3a和N-cadherin的mRNA及蛋白表达显著升高(P均<0.05),组间差距均具有统计学意义。结论黑素瘤细胞中miR-136水平较正常细胞降低,过度表达的miR-136通过下调Wnt信号传导途径,抑制黑素瘤细胞的EMT。Objective To explore the regulatory role of microrna-136(miR-136)in melanoma EMT based on the Wnt signaling pathway.Methods(negative control group)and B16 mouse melanoma cells were cultured routinely.B16 cells were divided into the blank group(no transfection sequence),the NC group(transfected with miR-136 NC),the miR-136 simulation group(transfected with miR-136 mimics)and the miR-136 inhibition group(transfected with miR-136 inhibitors)by liposome transfection.The expression of miR-136 in each group was detected by qRT-PCR.The differences in mRNA and protein expression of Wnt3 a,β-catenin,Ecadherin and N-cadherin in each group were detected by qRT-PCR and western-blot Compared with the negative control group,the expression of miR-136 in other groups was significantly reduced,and there was no significant difference between the blank group and the NC group(P>0.05).Compared with the blank group and the NC group,the expression of miR-136 in the miR-136 simulation group was significantly increased(P<0.05).The expression of miR-136 was significantly decreased in the miR-136 inhibition group(P<0.05).Compared with the control group,the mRNA and protein expressions of E-cadherin in other groups were significantly reduced(all P<0.05).The mRNA and protein expressions ofβ-catenin,Wnt3 a,and N-cadherin were significantly increased(all P<0.05).There was no significant difference between the blank group and the NC group(P>0.05).Compared with the blank group and NC group,mRNA and protein expressions of Ecadherin in miR-136 simulation group were significantly increased(all P<0.05),whereas those ofβ-catenin,Wnt3 a and N-cadherin were significantly decreased(all P<0.05).The mRNA and protein expressions of E-cadherin in the miR-136 inhibitor group were significantly decreased(all P<0.05),while those ofβ-catenin,Wnt3 a and N-cadherin were significantly increased(all P<0.05).The differences between groups were statistically significant.Conclusion The level of miR-136 in melanoma cells is lower than that in normal cells.Over-expr

关 键 词：microRNA-136 黑素瘤 WNT信号通路 EMT 

分 类 号：R739.5[医药卫生—肿瘤]