干扰HMGB1表达对膀胱癌细胞株恶性行为及其化疗敏感性的影响
发布时间：2019-04-21 12:53
【摘要】：目的:探讨高迁移率族蛋白1对膀胱癌细胞株的恶性生物学行为的作用以及其对膀胱癌化疗耐药的影响。方法:20例膀胱肿瘤以及相应癌旁组织,免疫组化方法检测癌和癌旁组织HMGB1表达差异;应用RNAi处理膀胱癌细胞株T24和BIU-87并分组;CCK8、流式细胞术、划痕实验和侵袭实验检测HMGB1敲低后对T24细胞增殖、周期、迁移以及侵袭能力的影响;使用共聚焦显微镜和透射电镜观察LC3荧光斑点;Western blot法检测相关蛋白的表达。结果:HMGB1在癌组织中的表达显著高于癌旁组织(P0.05);CCK8结果提示,相较于Blank组和NC组,HMGB1干扰组细胞增殖受抑制;流式细胞术提示敲低干扰组中G0/G1期细胞数量增加,且吉西他滨诱导的细胞凋亡增加;划痕测试法和Transwell侵袭试验显示敲低HMGB1后细胞迁移以及侵袭能力减弱;Western blot结果显示敲低HMGB1后E-cadherin蛋白表达上调,N-cadherin、vimentin、MMP-2、MMP-9、cyclinD1、c-Myc、β-catenin蛋白表达下调,同时可进一步增加吉西他滨诱导的caspase 3和PARP1的活化。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,也可通过促进自噬活化而参与化疗耐药的过程,使膀胱癌细胞对药物敏感性降低。
[Abstract]:Aim: to investigate the effect of high mobility group protein 1 on malignant biological behavior of bladder cancer cell line and its effect on chemotherapy resistance of bladder cancer cell line. Methods: the expression of HMGB1 was detected by immunohistochemical method in 20 cases of bladder tumors and corresponding paracancerous tissues, and the bladder cancer cell lines T24 and BIU-87 were treated with RNAi and divided into groups. CCK8, flow cytometry, scratch test and invasion assay were used to detect the effects of HMGB1 knockdown on proliferation, cycle, migration and invasion ability of T24 cells, and LC3 fluorescence spots were observed by confocal microscopy and transmission electron microscopy. The expression of related proteins was detected by Western blot. Results: the expression of HMGB1 in cancer tissues was significantly higher than that in non-cancerous tissues (P0.05), and the results of CCK8 showed that compared with Blank group and NC group, the proliferation of cells in HMGB1 interference group was inhibited. Flow cytometry showed that the number of G0/G1-phase cells increased and the apoptosis induced by gemcitabine increased in the knock-down group, and the migration and invasion ability of the cells decreased after HMGB1 knockdown by scratch assay and Transwell invasion test. The results of Western blot showed that the expression of E-cadherin protein was up-regulated after knock-down of HMGB1, and the expression of E-cadherin protein was down-regulated, and the expression of caspase-3 and PARP1 induced by gemcitabine was further increased by down-regulation of mRNA expression of MMP-2, MMP-9, cyclin D1, c-myc and 尾-catenin. Conclusion: HMGB1 can enhance the malignant biological behavior of bladder cancer cell line EMT by promoting the activation of autophagy and participate in the process of chemotherapy resistance, thus reducing the drug sensitivity of bladder cancer cell line.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.14
本文编号：2462219
[Abstract]:Aim: to investigate the effect of high mobility group protein 1 on malignant biological behavior of bladder cancer cell line and its effect on chemotherapy resistance of bladder cancer cell line. Methods: the expression of HMGB1 was detected by immunohistochemical method in 20 cases of bladder tumors and corresponding paracancerous tissues, and the bladder cancer cell lines T24 and BIU-87 were treated with RNAi and divided into groups. CCK8, flow cytometry, scratch test and invasion assay were used to detect the effects of HMGB1 knockdown on proliferation, cycle, migration and invasion ability of T24 cells, and LC3 fluorescence spots were observed by confocal microscopy and transmission electron microscopy. The expression of related proteins was detected by Western blot. Results: the expression of HMGB1 in cancer tissues was significantly higher than that in non-cancerous tissues (P0.05), and the results of CCK8 showed that compared with Blank group and NC group, the proliferation of cells in HMGB1 interference group was inhibited. Flow cytometry showed that the number of G0/G1-phase cells increased and the apoptosis induced by gemcitabine increased in the knock-down group, and the migration and invasion ability of the cells decreased after HMGB1 knockdown by scratch assay and Transwell invasion test. The results of Western blot showed that the expression of E-cadherin protein was up-regulated after knock-down of HMGB1, and the expression of E-cadherin protein was down-regulated, and the expression of caspase-3 and PARP1 induced by gemcitabine was further increased by down-regulation of mRNA expression of MMP-2, MMP-9, cyclin D1, c-myc and 尾-catenin. Conclusion: HMGB1 can enhance the malignant biological behavior of bladder cancer cell line EMT by promoting the activation of autophagy and participate in the process of chemotherapy resistance, thus reducing the drug sensitivity of bladder cancer cell line.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.14
【参考文献】
相关期刊论文 前2条
1 Zhe Liu;Ruixia Du;Jin Long;Kejian Guo;Chunlin Ge;Shulong Bi;Yuanhong Xu;;microRNA-218 promotes gemcitabine sensitivity in human pancreatic cancer cells by regulating HMGB1 expression[J];Chinese Journal of Cancer Research;2015年03期
2 Jing Zhang;Cang Liu;Ruiguang Hou;;Knockdown of HMGB1 improves apoptosis and suppresses proliferation and invasion of glioma cells[J];Chinese Journal of Cancer Research;2014年06期
，本文编号：2462219
本文链接：https://www.wllwen.com/yixuelunwen/mjlw/2462219.html
最近更新
教材专著
