摘要
目的观察葡萄糖调节蛋白94(GRP94)在膀胱肿瘤组织和细胞中的表达情况,并下调膀胱肿瘤细胞内GRP94表达,检测其对肿瘤细胞凋亡及迁移能力的影响,探讨其可能机制。方法选取35例膀胱尿路上皮癌组织和35例癌旁组织,应用免疫组化方法检测GRP94在两种组织中的表达,同时应用Western blot法检测GRP94在正常膀胱上皮细胞株和膀胱尿路上皮癌细胞株中的表达;设计合成可靶向结合GRP94序列的小干扰RNA(siGRP94),将其导入膀胱尿路上皮癌SW780细胞株,随后应用Western blot法检测SW780细胞内GRP94的表达情况;并分别应用流式细胞术检测凋亡率,Transwell法检测迁移能力;同时应用Western blot法检测细胞内caspase-9、Bcl-2、Bax和vimentin表达情况。结果膀胱尿路上皮癌组织中GRP94表达的平均光密度值明显高于癌旁组织(P<0.01),膀胱尿路上皮癌细胞株中GRP94表达明显高于正常膀胱上皮细胞株(P<0.01);siGRP94转染后48 h,SW780细胞内GRP94表达较空白组和阴性对照组明显下调(P<0.01),细胞凋亡率明显上升(P<0.01),同时迁移能力明显下降(P<0.01);凋亡调控蛋白中,caspase-9前体表达下调同时剪切体表达上调(P<0.01),Bcl-2表达下调同时Bax表达上调(P<0.01);细胞迁移相关蛋白波形蛋白表达明显下调(P<0.01)。结论GRP94在膀胱尿路上皮癌中呈高表达,下调其表达水平可诱导膀胱肿瘤细胞发生凋亡并抑制细胞迁移能力,GRP94可能具有成为膀胱肿瘤治疗靶基因的潜力。
Objective To observe the expression of glucose-regulated protein 94(GRP94)in bladder tumor tissues and cells,study the effect of GRP94 expression on cancer cell apoptosis and migration ability by down-regulating its expression in bladder tumor cells,and explore its possible mechanism.Methods Thirty-five cases of bladder urothelial carcinoma and 35 adjacent tissues were selected.The expressions of GRP94 in these 2 types of tissues we reexamined by immunohistochemistry.Meanwhile,Western blot was used to test the expression of GRP94 in normal bladder epithelial cell line and bladder urothelial carcinoma cell lines.Small interfering RNA targeting GRP94 sequence(siGRP94)was designed and synthesized,and then transfected into the bladder urothelial carcinoma SW780 cell line.The expression of GRP94 in SW780 cells was detected by Western blot.The apoptosis rate and the cell migration ability were detected by flow cytometry and Transwell chamber,respectively.Furthermore,the expressions of caspase-9,Bcl-2,Bax and vimentin were detected by Western blot.Results The mean optical density of GRP94 expression in bladder urothelial carcinoma was significantly higher than that in adjacent tissues(P<0.01),and the expression of GRP94 in bladder urothelial carcinoma cell line was obviously higher than that in normal bladder epithelial cell line(P<0.01).The expression of GRP94 in SW780 cells exhibited a significantly decrease compared to the blank group and the negative control group(P<0.01),and the apoptosis rate was significantly increased(P<0.01)with a significant decrease in the migration ability(P<0.01)at 48 hours after transfection of siGRP94.As for apoptosis-regulated proteins,the expression of caspase-9 precursor was down-regulated and the expression of splicing was up-regulated(P<0.01).The expression of Bcl-2 was down-regulated and the expression of Bax was up-regulated(P<0.01).Besides,significant down-regulation expression of cell migration associated protein vimentin was displayed(P<0.01).Conclusions GRP94 is over-expressed
作者
卢童
徐康
LU Tong;XU Kang(Department of Urology, First People's Hospital of Tianmen City, Tianmen 431700, China)
出处
《现代泌尿生殖肿瘤杂志》
2021年第6期353-358,共6页
Journal of Contemporary Urologic and Reproductive Oncology
关键词
膀胱肿瘤
葡萄糖调节蛋白94
凋亡
迁移
Bladder tumor
Glucose-regulated protein 94
Apoptosis
Migration
