摘要
目的:探讨敲低烷基化修复同源体3 (ALKBH3)对膀胱癌细胞生长、迁移与肿瘤血管生成的影响,并阐明其相关机制。方法:分别采用免疫组织化学和蛋白质印迹法检测膀胱上皮细胞癌和癌旁组织以及正常膀胱上皮细胞(SV-HRUC-1)和膀胱癌细胞(BIU87和T24)中ALKBH3蛋白表达水平。将BIU87和T24细胞分为sh-Con组(转染sh-Con)和sh-ALKBH3组(转染sh-ALKBH3)。采用CCK-8法、集落形成实验和Transwell小室实验分别检测2组细胞活性、集落数、迁移数和侵袭数。采用Transwell小室实验和小管形成实验分别检测2组细胞培养基诱导的人脐静脉内皮细胞(HUVEC)迁移数和小管形成率。构建异体移植瘤模型,采用游标卡尺定时检测2组瘤体体积,采用免疫组织化学检测瘤体组织中CD31蛋白表达水平。采用蛋白质印迹法检测体内和体外实验中2组细胞中血管内皮生长因子A (VEGF-A)蛋白表达水平。结果:与癌旁组织或SV-HRUC-1细胞比较,膀胱上皮细胞癌组织或膀胱癌细胞(BIU87和T24)中ALKBH3蛋白表达水平均明显升高(P<0.01)。体外实验,与sh-Con组比较,sh-ALKBH3组BIU87和T24细胞活性、迁移数和侵袭数以及其诱导的HUVEC迁移数和小管形成率均明显降低(P<0.01)。体内实验,与sh-Con组比较,sh-ALKBH3组瘤体体积减小(P<0.01),且瘤体组织中CD31蛋白表达水平明显降低(P<0.01)。与sh-Con组比较,在体内外实验中sh-ALKBH3组VEGF-A蛋白表达水平均明显降低(P<0.01)。结论:敲低ALKBH3能抑制膀胱癌细胞的生长、迁移、侵袭和肿瘤血管生成,其机制可能与下调VEGF-A表达有关。
Objective:To investigate the effect of alkylation repair homolog 3(ALKBH3)knockdown on the growth,migration and tumor angiogenesis of bladder cancer cells,and to elucidate its related mechanisms.Methods:The expression levels of ALKBH3 protein in the urothelial cell carcinoma and paracarcinoma tissues,normal bladder epithelial cells(SV-HRUC-1),and bladder cancer cells(BIU87 and T24)were detected by immunohistochemistry and Western blotting method,respectively.The BIU87 and T24 cells were divided into sh-Con group(transfected with sh-Con)and sh-ALKBH3 group(transfected with sh-ALKBH3).The cell viabilities and the number of colony,migration and invasion in two groups were detected by CCK-8 method,colony formation experiment and Transwell chamber assay,respectively.The number of migration cells and the tubule formation rate of human umbilical vein endothelial cells(HUVEC)induced by cell culture medium in two groups were detected by Transwell chamber and tubule formation assays,respectively.After the xenograft tumor model was constructed,the tumor volumes in two groups were measured regularly with vernier caliper,and the expression levels of CD31 protein in the tumor tissue in two groups were detected by immunohistochemistry.The expression levels of vascular endothelial growth factor-A(VEGF-A)protein in in vivo and in vitro experiments in two groups were detected by Western blotting method.Results:Compared with para-carcinoma tissue or SVHRUC-1 cells,the expression levels of ALKBH3 proteins in urothelial cell carcinoma tissue and bladder cancer cells(BIU87 and T24)were significantly increased(P<0.01).The in vitro experiment results showed that compared with sh-Con group,the cell activities,the number of migration and invasion BIU87 and T24 cells,as well as the number of induced migration HUVEC and the tubule formation rate of HUVEC in sh-ALKBH3 group were significantly decreased(P<0.01).The vivo experiment results showed that compared with sh-Con group,the tumor volume in sh-ALKBH3 group was significantly reduced,and
作者
赵琪
何长海
王志
王雪峰
刘晓飞
ZHAO Qi;HE Changhai;WANG Zhi;WANG Xuefeng;LIU Xiaofei(Department of Urinary Surgery,Nanyang First People’s Hospital,Nanyang 473010,Henan Province,China)
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2021年第2期397-406,共10页
Journal of Jilin University：Medicine Edition
基金
河南省卫计委科研基金项目(201602014)。
