干扰HOXA9基因表达对BTT739细胞增殖、迁移和侵袭的影响  

Effects of HOXA9 silencing on the proliferation and invasion of mouse bladder cancer cell line BTT739 cells

作　　者：冉崇君 张鹏[1] 陈毅 郭江 种袁佳 张克勤 梁培禾[1] RAN Chongjun;ZHANG Peng;CHEN Yi;GUO Jiang;CHONG Yuanjia;ZHANG Keqin;LIANG Peihe(Department of Urology,Second Affiliated Hospital of Chongqing Medical University,Chongqing 400000,China)

机构地区：[1]重庆医科大学附属第二医院泌尿外科,400000

出　　处：《免疫学杂志》2023年第5期388-395,共8页Immunological Journal

基　　金：2017年重庆市第二批科技计划(cstc2017jcyjAX0435);2019年重庆市留学人员回国创业创新支持计划(CX2019146)。

摘　　要：目的探讨同源盒基因HOXA9表达对小鼠膀胱尿路上皮癌细胞系BTT739细胞增殖、迁移和侵袭的影响。方法免疫组化检测膀胱尿路上皮癌以及癌旁正常组织临床标本中HOXA9基因的表达情况;采用sh-RNA技术干扰BTT739细胞中HOXA9基因的表达后,进行克隆形成、CCK8、流式细胞术、Transwell和划痕等实验检测BTT739细胞增殖、凋亡、迁移以及侵袭等生物学特性的变化,并通过Western blot检测上皮间质转化(epithelial to mesenchymal transition,EMT)特征因子E-cadherin、Vi⁃mentin和Snail的表达。结果膀胱尿路上皮癌组织中HOXA9基因的表达高于癌旁正常组织。sh-RNA敲低BTT739细胞中的HOXA9基因后,细胞增殖、迁移和侵袭能力显著下降,而细胞早期凋亡百分率增高,G1、G2期比例下降,S期升高;此外,EMT特征因子E-cadherin表达增强,Vimentin和Snail的表达下降。结论HOXA9基因具有促进小鼠膀胱尿路上皮癌细胞系BTT739细胞增殖、迁移和侵袭能力的作用,是膀胱尿路上皮癌潜在的免疫治疗靶点,其机制可能涉及EMT机制。This study was designed to investigate the effects of HOXA9 expression on the proliferation,migration and invasion of mouse bladder cancer cell BTT739.Before the cell experiment,we detected the expression of HOXA9 gene in the samples of bladder urothelial cancer and adjacent normal tissues from surgical operation by immunohistochemistry.Then in later experiments,the expression of HOXA9 gene in BTT739 cells was knockdown by sh-RNA and the proliferation,apoptosis,migration and invasion were detected by clone formation,flow cytometry,CCK8,Transwell and scratch test,respectively.The expression of epithelial to mesenchymal transition(EMT)characteristic factors,including E-cadherin,Vimentin and Snail,were measured by Western blotting.The results showed that the expression of HOXA9 gene in bladder cancer tissues was much higher than that in the adjacent normal tissues.Both the mRNA and protein expression of HOXA9 gene in BTT739 cells were successfully knocked down,which led to a significant reduction in cells proliferation,migration and invasion,a significant increase in the early apoptosis percentage and the proportion of S phase,while a significant decrease in the proportion of G1 and G2 phase.Meanwhile,EMT characteristic factor E-cadherin expression was enhanced while the expression of Vimentin and Snail were decreased.In conclusion,HOXA9 gene may be an oncogene with the function of promoting BTT739 cells proliferation,migration and invasion,and the mechanism of this process involves EMT.Therefore,HOXA9 is expect to become a potential target for immunotherapy of bladder cancer.

关 键 词：HOXA9 膀胱尿路上皮癌 BTT739细胞 凋亡 免疫治疗 

分 类 号：R737.14[医药卫生—肿瘤]