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长基因间非编码RNA 00689靶向miR⁃876⁃5p调控膀胱癌T24细胞增殖和凋亡的机制 认领

Effect of LINC00689 on proliferation and apoptosis of bladder cancer T24 cells through targeting miR⁃876⁃5p
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摘要 目的探讨长基因间非编码RNA 00689(LINC00689)对膀胱癌细胞T24增殖、凋亡的影响和可能机制。方法实时荧光定量PCR(RT⁃qPCR)分析膀胱癌组织、癌旁组织、T24细胞以及正常膀胱上皮细胞SV⁃HUC⁃1中LINC00689和miR⁃876⁃5p表达。双荧光素酶报告基因和RT⁃qPCR验证LINC00689对miR⁃876⁃5p的调控作用。将T24细胞分为si⁃NC组、si⁃LINC00689组、miR⁃NC组、miR⁃876⁃5p组、si⁃LINC00689+anti⁃miR⁃NC组和si⁃LINC00689+anti⁃miR⁃876⁃5p组。运用细胞计数试剂盒(CCK⁃8)、流式细胞术分析细胞活力和凋亡,蛋白质免疫印记(Western blot)分析细胞周期素D1(CyclinD1)、p21、B细胞淋巴瘤2(Bcl⁃2)和Bcl相关X蛋白(Bax)表达。结果膀胱癌组织、T24细胞中LINC00689表达升高,miR⁃876⁃5p表达降低(P<0.05)。LINC00689负调控miR⁃876⁃5p表达。与si⁃NC组比较,si⁃LINC00689组细胞活力、CyclinD1和Bcl⁃2蛋白表达降低,凋亡率、p21和Bax蛋白升高(P<0.05)。与miR⁃NC组比较,miR⁃876⁃5p组细胞活力、CyclinD1和Bcl⁃2蛋白表达降低,凋亡率、p21和Bax蛋白升高(P<0.05)。与si⁃LINC00689+anti⁃miR⁃NC组比较,si⁃LINC00689+anti⁃miR⁃876⁃5p组细胞活力、CyclinD1和Bcl⁃2蛋白表达升高,凋亡率、p21和Bax蛋白降低(P<0.05)。结论抑制LINC00689通过负调控miR⁃876⁃5p能够降低膀胱癌T24细胞的增殖能力,并促进细胞凋亡。 Objective To investigate the effect of long intergenic non⁃coding RNA 00689(LINC00689)on the proliferation and apoptosis of bladder cancer cell T24 and its possible mechanism.Methods Real⁃time fluorescence quantitative PCR(RT⁃qPCR)was used to analyze the expression of LINC00689 and miR⁃876⁃5p in bladder cancer tissues,adjacent tissues,T24 cells and normal bladder epithelial cells SV⁃HUC⁃1.Dual luciferase reporter gene and RT⁃qPCR were applied to verify the regulation effect of LINC00689 on miR⁃876⁃5p.T24 cells were divided into si⁃NC group,si⁃LINC00689 group,miR⁃NC group,miR⁃876⁃5p group,si⁃LINC00689+anti⁃miR⁃NC group and si⁃LINC00689+anti⁃miR⁃876⁃5p group.Cell counting kit(CCK⁃8)and flow cytometry were used to analyze cell viability and apoptosis and western blot was used to analyze expression of cyclin D1(CyclinD1),p21,B cell lymphoma 2(Bcl⁃2)and Bcl⁃related X protein(Bax).Results The expression of LINC00689 was increased in bladder cancer tissues and T24 cells,but that of miR⁃876⁃5p was decreased(P<0.05).LINC00689 negatively regulated miR⁃876⁃5p expression.Compared with those in si⁃NC group,cell viability,expression of CyclinD1 but Bcl⁃2 proteins were reduced,and apoptosis rate,expression of p21 and Bax proteins were increased in si⁃LINC00689 group(P<0.05).Compared with those in miR⁃NC group,cell viability,expression of CyclinD1 and Bcl⁃2 pro⁃teins were reduced,but apoptosis rate,expression of p21 and Bax proteins were increased in miR⁃876⁃5p group(P<0.05).Compared with those in si⁃LINC00689+anti⁃miR⁃NC group,cell viability,expression of CyclinD1 and Bcl⁃2 proteins were increased,but apoptosis rate,expression of p21 and Bax proteins were reduced in si⁃LINC00689+anti⁃miR⁃876⁃5p group(P<0.05).Conclusion Inhibiting LINC00689 can reduce the proliferation of bladder cancer T24 cells and promote apoptosis by negatively regulating miR⁃876⁃5p.
作者 杨文博 许振丹 耿帅 YANG Wenbo;XU Zhendan;GENG Shuai(Department of Urinary Surgery,Xinxiang Central Hospital,Xinxiang 453000,China)
出处 《实用医学杂志》 CAS 北大核心 2021年第2期195-200,共6页 The Journal of Practical Medicine
基金 2018年度河南省医学科技攻关计划项目(编号:201820938)。
关键词 LINC00689 膀胱癌 细胞增殖 凋亡 miR⁃876⁃5p LINC00689 bladder cancer cell proliferation apoptosis miR⁃876⁃5p
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