siRNA干扰HMGB1表达对膀胱癌T24株细胞恶性生物学行为的影响及其可能的机制

发布时间：2018-05-08 06:27

本文选题：高迁移率族蛋白 + 膀胱癌细胞　；参考：《中国肿瘤生物治疗杂志》2017年03期

【摘要】：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),P0.05]。与NC组和Blank组相比,si HMGB1组细胞增殖受抑制(P0.05);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(P0.05)以及侵袭[穿膜细胞数:(16.33±1.45)vs(35.00±1.53)、(34.00±2.08)个,均P0.05]能力减弱;Western blotting结果显示敲低后E-钙黏着蛋白表达上调(P0.05),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P0.05)。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。
[Abstract]:Objective: to investigate the effects of high mobility group protein 1(high-mobility group box 1 HMGB1 on the proliferation, apoptosis and malignant biological behavior of bladder cancer cell line T24. Methods: from December 2014 to January 2016, 20 cases of bladder cancer and its adjacent tissues were collected from the urology ward of the first affiliated Hospital of Chongqing Medical University. The expression of HMGB1 in bladder cancer and adjacent tissues was detected by immunohistochemical method. T24 cells were treated with RNAi and divided into blank control group, negative control group and interference group. Flow cytometry, scratch test and Transwell invasion assay were used to detect the proliferation, apoptosis and cycle of T24 cells after HMGB1 knockout. The expression of HMGB1 in different bladder cancer cell lines BIU-87 and T24 cells was detected by Western blotting and the effect of knock down HMGB1 on the expression of proteins associated with malignant biological behavior of T24 cells was evaluated by Western blotting. Results: the expression of HMGB1 in bladder cancer tissues was higher than that in paracancerous tissues [67.33 卤4.91)vs(12.00 卤3.79 P 0.05]. Compared with NC group and Blank group, cell proliferation was inhibited in HMGB1 group (P 0.05), apoptosis rate was increased and cell cycle was blocked in G0/G1 phase after knockdown with low HMGB1 by flow cytometry. The ability of cell migration (P0.05) and invasion (16.33 卤1.45)vs(35.00 卤1.53 卤34.00 卤2.08) of cell migration after knockdown (P0.05) was decreased. The results of Western blotting showed that the expression of E-cadherin up-regulated the expression of P0.05 N-cadherin and vimentin after knockout. The expression of matrix metalloproteinase (MMP), MMP- 2, MMP-9, and cyclin D1, c-Myc, 尾-binding protein were down-regulated (all P 0.05). ConclusionHMGB1 can enhance the malignant biological behavior of bladder cancer cells by promoting EMT, and its mechanism may be mediated by 尾 -integrin signaling pathway.
【作者单位】：重庆医科大学附属第一医院泌尿外科;重庆医科大学附属第一医院重庆市分子肿瘤及表观遗传学重点实验室;
【基金】：国家自然科学基金资助项目(No.81372758)~~
【分类号】：R737.14

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