敲低m6A甲基转移酶WTAP对皮肤鳞癌细胞增殖、迁移、侵袭能力的影响  

Effects of knocking down m6A methyltransferase WTAP on proliferation,migration,and invasion of cutaneous squamous-cell carcinoma cells

作　　者：黄志顺 霍然 HUANG Zhishun;HUO Ran(Department of Plastic Surgery,Shandong Provincial Hospital Affiliated to Shandong First Medical University,Jinan 250021,China)

机构地区：[1]山东第一医科大学附属省立医院整形美容外科,济南250021 [2]山东第一医科大学医学科技创新中心 [3]山东大学附属省立医院整形美容外科

出　　处：《山东医药》2023年第17期33-37,共5页Shandong Medical Journal

摘　　要：目的通过敲低N6-甲基腺嘌呤(m6A)甲基转移酶Wilms肿瘤蛋白1相关蛋白(WTAP)在皮肤鳞癌(cSCC)细胞中的表达,探讨WTAP对cSCC细胞增殖,迁移和侵袭能力的影响。方法选取人cSCC细胞A431、SCL-1及正常人表皮角质形成细胞HaCaT,Western blotting法及qRT-PCR法分别检测各细胞中WTAP蛋白及mRNA表达。选取WTAP蛋白及mRNA表达最高的A431细胞转染WTAP敲低慢病毒shWTAP-1、shWTAP-2、shWTAP-3及阴性对照慢病毒shNC,使用qRT-PCR与Western blotting验证转染效率。选取敲低差异最显著的shWTAP-3转染细胞作为WTAP敲低组,shNC转染细胞作为对照组。采用比色法检测细胞m6A表达,CCK-8法及克隆形成实验检测细胞增殖能力,Transwell实验检测细胞侵袭能力,细胞划痕实验检测细胞迁移能力。结果WTAP敲低组m6A表达低于对照组,各时点细胞OD值均低于对照组,克隆形成数少于对照组,穿膜细胞数、细胞迁移率低于对照组(P均<0.05)。结论敲低WTAP可抑制cSCC细胞的增殖、侵袭和迁移,该作用可能通过下调m6A表达来实现。Objective By knocking down the expression of N6-methyladenine(m6A)methyltransferase Wilms'tumor 1-associated protein(WTAP)in cutaneous squamous-cell carcinoma(cSCC)cells,the effects of WTAP on the proliferation,migration,and invasion of cSCC cells were investigated.Methods Human cSCC cells A431 and SCL-1 and normal human epidermal keratinocytes HaCaT were selected.Western blotting and qRT-PCR were used to detect the protein and mRNA expression levels of WTAP in each cell.A431 cells with the highest expression of WTAP protein and mRNA were transfected with WTAP knockdown lentivirus shWTAP-1,shWTAP-2,shWTAP-3 and negative control lentivirus shNC,and the transfection efficiency was verified by qRT-PCR and Western blotting.The shWTAP-3 transfected cells with the most significant knockdown difference were selected as the WTAP knockdown group,and the shNC transfected cells were selected as the control group.The expression of m6A was detected by colorimetric assay,the proliferation ability of cells was detected by CCK-8 assay and colony formation assay,the invasion ability of cells was detected by Transwell assay,and the migration ability of cells was detected by cell scratch assay.Results The expression of m6A in the WTAP knockdown group was lower than that in the control group,OD value at each time point was lower than that in the control group,the number of colony formation was smaller than that in the control group,and the number of transmembrane cells and cell migration rate were lower than those in the control group(all P<0.05).Conclusions Knocking down WTAP can inhibit the proliferation,invasion and migration of cSCC cells,which may be achieved by down-regulating the expression of m6A.

关 键 词：Wilms肿瘤蛋白1相关蛋白 N6-甲基腺嘌呤 细胞增殖 细胞侵袭 细胞迁移 皮肤鳞状细胞癌 

分 类 号：R739.5[医药卫生—肿瘤]