OBJECTIVE:This study aimed to investigate the effect of circular RNA itchy E3 ubiquitin protein ligase on cell proliferation and apoptosis and to explore its target micro-RNAs in prostate cancer cells. METHODS:Circular RNA itchy E3 ubiquitin protein ligase expression in human prostate cancer cells and normal prostate epithelial cells was determined by real time-quantitative polymerase chain reaction assay. Circular RNA itchy E3 ubiquitin protein ligase overexpression plasmids (circular RNA itchy E3 ubiquitin protein ligase(+) group and control overexpression plasmids group were transfected with PC-3 cells. Rescue experiment was performed by transfection of circular RNA itchy E3 ubiquitin protein ligase overexpression and micro-197 overexpression plasmids (circular RNA itchy E3 ubiquitin protein ligase overexpression plasmids/micro RNA (+) group) into PC-3 cells. Cell Counting Kit-8 and annexin V/propidium iodide assays were conducted to evaluate cell proliferation and apoptosis, respectively. Western blot was performed to determine the expressions of apoptotic-related markers. RESULTS:Circular RNA itchy E3 ubiquitin protein ligase expression was decreased in DU 145, 22RV1, VCaP, and PC-3 cells compared to RWPE cells. In PC-3 cells, cell proliferation rate was reduced in circular RNA itchy E3 ubiquitin protein ligase overexpression plasmids group compared to control overexpression plasmids group at 48 hours and 72 hours. Cell apoptosis rate was elevated in circular RNA itchy E3 ubiquitin protein ligase overexpression plasmids group compared to control overexpression plasmids group at 48 hours, and Western blot showed the similar results. Micro RNA-197 but not micro RNA-31 or micro RNA-432 was the target micro-RNA of circular RNA itchy E3 ubiquitin protein ligase. In rescue experiments, cell proliferation rate was elevated, but apoptosis rate was reduced in circular RNA itchy E3 ubiquitin protein ligase overexpression plasmids/micro RNA (+) group compared to circular RNA itchy E3 ubiquitin protein ligase overexpression plasmids group, indicating that circular RNA itchy E3 ubiquitin protein ligase upregulation inhibited cell proliferation but promoted apoptosis through downregulating micro RNA-197. CONCLUSION:Circular RNA itchy E3 ubiquitin protein ligase upregulation suppresses cell proliferation but promotes apoptosis through targeting micro RNA-197 in prostate cancer. Our study may provide a new insight for the treatment of prostate cancer.

译文

目的:本研究旨在研究环状RNA痒E3泛素蛋白连接酶对前列腺癌细胞增殖和凋亡的作用,并探讨其靶微RNA的表达。
方法:采用实时定量聚合酶链反应法测定人前列腺癌细胞和正常前列腺上皮细胞中的环状RNA发痒的E3泛素蛋白连接酶的表达。环状RNA痒E3泛素蛋白连接酶过表达质粒(环状RNA痒E3泛素蛋白连接酶()组和对照过表达质粒组分别用PC-3细胞转染,通过转染环状RNA痒E3泛素蛋白连接酶过表达和微量表达来进行抢救实验-197过度表达质粒(环状RNA痒E3泛素蛋白连接酶过度表达质粒/ micro RNA()组)进入PC-3细胞,进行Cell Counting Kit-8和Annexin V /碘化丙啶检测,分别评估细胞增殖和凋亡。进行蛋白质印迹以确定凋亡相关标志物的表达。
结果:与RWPE细胞相比,DU 145、22RV1,VCaP和PC-3细胞中环状RNA发痒的E3泛素蛋白连接酶表达降低。在PC-3细胞中,与对照组的过表达质粒组相比,在48小时和72小时时,环状RNA发痒的E3泛素蛋白连接酶过表达质粒组的细胞增殖速率降低。与对照过表达质粒组相比,在48小时时,环状RNA瘙痒E3泛素蛋白连接酶过表达质粒组的细胞凋亡率升高,并且Western印迹显示了相似的结果。微小RNA-197但不是微小RNA-31或微小RNA-432是环状RNA发痒的E3泛素蛋白连接酶的目标微小RNA。在急救实验中,与环状RNA瘙痒E3泛素蛋白连接酶过表达质粒组相比,环状RNA瘙痒E3泛素蛋白连接酶过表达质粒组的细胞增殖速率提高,但凋亡率降低,表明环状RNA瘙痒E3泛素蛋白连接酶过表达质粒组泛素蛋白连接酶上调抑制细胞增殖,但通过下调micro RNA-197促进细胞凋亡。
结论:环状RNA发痒的E3泛素蛋白连接酶上调通过靶向微小RNA-197靶向前列腺癌抑制细胞增殖,但促进细胞凋亡。我们的研究可能为前列腺癌的治疗提供新的见解。

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