长链非编码RNA HOXA末端转录本反义RNA在膀胱癌中的表达及其对膀胱癌细胞增殖、迁移与侵袭的影响  

Expression of long-stranded noncoding RNA HOXA transcript at the distal tip in bladder cancer and its effect on proliferation, migration and invasion of bladder cancer cells

作　　者：汤利 郭永连[1] 陈琳[1] 李国灏[1] 张伟[1] Tang Li;Guo Yonglian;Chen Lin;Li Guohao;Zhang Wei(Department of Urology, the Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, China)

机构地区：[1]华中科技大学同济医学院附属武汉市中心医院泌尿外科,武汉430014

出　　处：《中华实验外科杂志》2019年第4期682-684,共3页Chinese Journal of Experimental Surgery

基　　金：湖北省卫生与计划生育委员会基金(WJ2017M184).

摘　　要：目的探讨长链非编码RNA HOXA末端转录本反义RNA(HOTTIP)在膀胱癌细胞中的表达,及其对膀胱癌细胞增殖、迁移与侵袭的影响及其作用机制。方法采用反转录聚合酶链反应(RT-PCR)检测HOTTIP在癌旁组织和膀胱肿瘤组织、正常上皮细胞和膀胱癌细胞间的表达差异;细胞计数试剂盒(CCK-8)观察膀胱癌细胞的增殖情况;流式细胞术检测膀胱癌细胞的细胞周期进程;运用Transwell和划痕愈合实验观察细胞侵袭和迁移能力;Western blot检测细胞中基质金属蛋白酶(MMPs)及其抑制剂(TIMPs)蛋白表达水平。结果HOTTIP在膀胱肿瘤组织中的表达(1.943±0.251)明显高于癌旁组织中的表达(1.002±0.091),差异有统计学意义(P<0.05);人源正常上皮细胞(SV-HUC-1)中HOTTIP的表达明显低于5637和T24中的表达(1.003±0.092比2.241±0.306和1.692±0.203),差异有统计学意义(P<0.05)。HOTTIP干扰组(si-HOTTIP)和阴性转染组(si-NC)干预5637和T24细胞48 h后,5637(0.881±0.182比1.824±0.193)和T24(0.623±0.094比1.332±0.153)细胞增殖能力显著降低(P<0.05),细胞周期均被阻滞在G0~G1期,细胞的迁移距离及穿膜率均降低,MMP-2和MMP-9蛋白表达水平均降低,TIMP1和TIMP2蛋白表达水平增加。结论长链非编码RNA HOTTIP在膀胱肿瘤组织和细胞中下调,通过降低MMPs和升高TIMPs促进膀胱癌细胞增殖、迁移和侵袭。Objective To investigate the expression of long-stranded non-coding RNA HOXA transcript at the distal tip (HOTTIP)(lncRNA HOPTTIP) in bladder cancer cells (BCC) and its effect on proliferation, migration and invasion of BCC and potential mechanisms. Methods Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the expression differences of HOTTIP between normal tissue and bladder cancer tissues, normal cell and bladder cancer cell. Cell counting kit-8 (CCK-8) was used to observe the proliferation levels of BCC. Cell cycle progression of BCC was detected by flow cytometry. Cell invasion and metastasis were observed by transwell and wound healing experiments. Western blotting assay was used to detect the protein expressions of matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinase (TIMPs). Results The expression level of HOTTIP in BCC (1.943±0.251) was significantly higher than that of paracancerous tissue (1.002±0.091), and the difference was statistically significant (P<0.05);The expression of HOTTIP in human normal epithelial cells (SV-HUC-1) was significantly lower than that of 5637 and T24 (1.003±0.092 vs. 2.241±0.306 and 1.692±0.203), with significant difference (P<0.05). After si-HOTTIP and si-NC intervention 48h on 5637 and T24 cells, the proliferation of 5637 (0.881±0.182 vs. 1.824±0.193) and T24 (0.623±0.094 vs. 1.332±0.153) cells decreased significantly (P<0.05), the cell cycle were stuck into G0/G1 phase, rate of invasion and migration were reduced, MMP-2 and MMP-9 protein expression levels decreased, TIMP1 and TIMP2 protein expression levels increased. Conclusion Long-stranded non-coding RNA (LncRNA) HOTTIP is down-regulated in bladder cancer tissues and cells, and promotes the proliferation, migration and invasion of BCC by decreasing MMPs and increasing TIMPs.

关 键 词：长链非编码RNAHOXA末端转录本反义RNA 膀胱癌 增殖 迁移 侵袭 

分 类 号：R737.14[医药卫生—肿瘤]