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目的:探讨高迁移率族蛋白1(high-mobility group box 1,HMGB1)对膀胱癌T24株细胞的增殖、凋亡及恶性生物学行为的影响及其潜在作用机制。方法:收集2014年12月至2016年1月期间重庆医科大学附属第一医院泌尿外科病区手术切除的20例膀胱癌以及相应癌旁组织,免疫组化方法检测膀胱癌和癌旁组织HMGB1表达差异;应用RNAi处理T24细胞并分为空白对照组(Blank)、阴性对照组(NC)和干扰组(si HMGB1);CCK-8、流式细胞术、划痕实验和Transwell侵袭实验分别检测HMGB1敲低后对T24细胞增殖、凋亡、周期、迁移以及侵袭能力的影响;Western blotting检测不同膀胱癌细胞株BIU-87和T24细胞的HMGB1表达水平以及敲低HMGB1对T24细胞恶性生物学行为相关蛋白的影响。结果:与癌旁组织相比,HMGB1在膀胱癌组织处于高表达状态[(67.33±4.91)vs(12.00±3.79),P<0.05]。与NC组和Blank组相比,si HMGB1组细胞增殖受抑制(P<0.05);流式细胞术提示敲低HMGB1后细胞凋亡率增加,细胞周期阻滞在G0/G1期;划痕实验及Transwell侵袭实验显示,敲低HMGB1后细胞迁移(P<0.05)以及侵袭[穿膜细胞数:(16.33±1.45)vs(35.00±1.53)、(34.00±2.08)个,均P<0.05]能力减弱;Western blotting结果显示敲低后E-钙黏着蛋白表达上调(P<0.05),N-钙黏着蛋白、波形蛋白、基质金属蛋白酶(metrix metalloproteinase,MMP)-2、MMP-9、细胞周期蛋白D1、c-Myc、β-联蛋白表达下调(均P<0.05)。结论:HMGB1可通过促进膀胱癌细胞EMT进而增强其恶性生物学行为,其机制可能是通过β-联蛋白信号通路介导。
Objective: To investigate the effects of high mobility group box 1 (HMGB1) on the proliferation, apoptosis and malignant behavior of bladder cancer T24 cells and its potential mechanism. Methods: Twenty cases of bladder cancer and corresponding paracancerous tissues resected in Department of Urology, the First Affiliated Hospital of Chongqing Medical University from December 2014 to January 2016 were collected. The expression of HMGB1 in bladder cancer and paracancerous tissues was detected by immunohistochemistry T cells were treated with RNAi and divided into blank control group, negative control group (NC) and interference group (si HMGB1); CCK-8, flow cytometry, scratch test and Transwell invasion assay were used to detect the expression of HMGB1 The effect of knockdown on the proliferation, apoptosis, cycle, migration and invasion ability of T24 cells was detected by Western blotting. The expression of HMGB1 in different bladder cancer cell lines BIU-87 and T24 and the knockdown of HMGB1 were detected by Western blotting The impact of protein. Results: HMGB1 was highly expressed in bladder cancer tissues compared with adjacent tissues (67.33 ± 4.91 vs 12.00 ± 3.79, P <0.05). Compared with NC group and Blank group, the proliferation of si HMGB1 group was inhibited (P <0.05). Flow cytometry indicated that the apoptosis rate of HMGB1 knockdown group was increased and the cell cycle arrest was in G0 / G1 phase. Scratch experiment (P <0.05) and invasion [cell number per cell: (16.33 ± 1.45) vs (35.00 ± 1.53), (34.00 ± 2.08), P <0.05] (P <0.05). N-Cadherin, vimentin, metrix metalloproteinase (MMP) -2, MMP-9, cyclin D1, c-Myc and β-catenin were down-regulated (all P <0.05). CONCLUSION: HMGB1 can enhance its malignant biological behavior by promoting EMT of bladder cancer cells. Its mechanism may be mediated by β-catenin signaling pathway.