标题：lncRNA LUCAT1对膀胱癌细胞增殖、凋亡的影响及其机制研究

      作者：王辉 1，田伟 2，杨建兵 1，李占琦 1，高卫军 1    陕西省核工业二一五医院泌尿外科 1、病理科 2，陕西 咸阳 712000

      卷次： 2020年31卷22期

      【摘要】 目的 探讨 lncRNA LUCAT1对膀胱癌细胞的增殖和凋亡的影响以及潜在的作用机制。方法 培养正常膀胱上皮细胞SV-HUC-1和膀胱癌细胞 J82、5637、BIU-87，将膀胱癌细胞 5637随机分为 control组、si-NC组，si-LUCAT1组、pcDNA-NC组、pcDNA-LUCAT1组、si-LUCAT1+anti-miR-NC组、si-LUCAT1+anti-miR-493-5p组。qRT-PCR法检测miR-493-5p和LUCAT1表达水平；Western Blot检测蛋白表达；MTT法检测细胞活性；流式细胞术检测细胞凋亡；双荧光素酶报告实验验证LUCAT1和miR-493-5p的靶向关系。结果 与膀胱上皮细胞SV-HUC-1相比，膀胱癌细胞 J82、5637、BIU-87中LUCAT1的表达水平明显升高，miR-493-5p表达水平明显降低，差异均有统计学意义(P<0.05)；与 si-NC组相比，si-LUCAT1组细胞活性降低，凋亡率升高，CyclinD1表达水平降低，Caspase-3表达水平升高，p-JAK2和p-STAT蛋白的表达水平降低，差异均有统计学意义(P<0.05)；miR-493-5p mimics与LUCAT1野生型报告质粒共转染的细胞荧光素酶活性降低，差异有统计学意义(P<0.05)；而miR-493-5p mimics与LUCAT1突变型报告质粒共转染的细胞荧光素酶活性无显著变化，差异无统计学意义(P>0.05)；与si-LUCAT1+anti-miR-NC组相比，si-LUCAT1+anti-miR-493-5p组细胞活性升高，细胞凋亡率降低，Cyclin D1表达水平升高，Caspase-3表达水平降低，p-JAK2和p-STAT蛋白表达水平升高，差异均有统计学意义(P<0.05)。结论 lncRNALUCAT1可抑制膀胱癌细胞增殖，促进其凋亡，其机制可能与miR-493-5p及 JAK/STAT信号通路有关。
      【关键词】 lncRNALUCAT1；miR-493-5p；JAK/STAT信号通路；膀胱癌；增殖；凋亡；作用机制
      【中图分类号】 R737.14 【文献标识码】 A 【文章编号】 1003—6350（2020）22—2857—05

Effect of lncRNA LUCAT1 on the proliferation and apoptosis of bladder cancer cells and its mechanism.
WANGHui 1, TIAN Wei 2, YANG Jian-bin 1, LI Zhan-qi 1, GAO Wei-jun 1. Department of Urology Surgery 1, Department of Pathology 2,No 215 Hospital of Shaanxi Nuclear Industry, Xianyang 712000, Shaanxi, CHINA
【Abstract】 Objective To investigate the effect of lncRNA LUCAT1 on the proliferation and apoptosis of blad-der cancer cells and its potential mechanism. Methods Culture normal bladder epithelial cells SV-HUC-1 and bladdercancer cells J82, 5637, BIU-87, and bladder cancer cells 5637 were randomly divided into the control group, si-NCgroup, si-LUCAT1 group, pcDNA-NC group, pcDNA-LUCAT1 group, si-LUCAT1+anti-miR-NC group, si-LUCAT1+anti-miR-493-5p group. qRT-PCR method was used to detect the expression of miR-493-5p and LUCAT1; Western blotwas used to detect protein expression; MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) methodwas used to detect cell viability; flow cytometry was used to detect apoptosis; dual luciferase report experiment was usedto verify the targeting relationship between LUCAT1 and miR-493-5p. Results Compared with bladder epithelial cellsSV-HUC-1, the expression level of LUCAT1 in bladder cancer cells J82, 5637, and BIU-87 was significantly increased,and the expression level of miR-493-5p was significantly reduced, with statistically significant differences (all P<0.05).Compared with the si-NC group, the cell activity of the si-LUCAT1 group was decreased, the apoptosis rate was in-creased, the expression level of CyclinD1 was decreased, the expression level of Caspase-3 was increased, and the ex-pression levels of p-JAK2 and p-STAT protein were decreased, with statistically significant differences (all P<0.05); theluciferase activity of cells co-transfected with miR-493-5p mimics and LUCAT1 wild-type reporter plasmid was de-creased, and the difference was statistically significant (P<0.05); the luciferase activity of cells co-transfected withmiR-493-5p mimics and LUCAT1 mutant reporter plasmid did not change significantly, and the difference was not sta-tistically significant (P>0.05); compared with the si-LUCAT1+anti-miR-NC group, in the si-LUCAT1+anti-miR-493-5pgroup, cell viability was increased, cell apoptosis rate was decreased, cyclin D1 expression level was increased, cas-pase-3 expression level was decreased, and p-JAK2 and p-STAT protein expression levels were increased, with statisti-cally significant differences (all P<0.05). Conclusion lncRNA LUCAT1 can inhibit the proliferation and promote theapoptosis of bladder cancer cells, and its mechanism may be related to miR-493-5p and JAK/STAT signaling pathway.
      【Key words】 LncRNA LUCAT1; miR-493-5p; JAK/STAT signaling pathway; Bladder cancer; Proliferation;Apoptosis; Mechanism