【摘要】：目的:探討高遷移率族蛋白1對(duì)膀胱癌細(xì)胞株的惡性生物學(xué)行為的作用以及其對(duì)膀胱癌化療耐藥的影響。方法:20例膀胱腫瘤以及相應(yīng)癌旁組織,免疫組化方法檢測癌和癌旁組織HMGB1表達(dá)差異;應(yīng)用RNAi處理膀胱癌細(xì)胞株T24和BIU-87并分組;CCK8、流式細(xì)胞術(shù)、劃痕實(shí)驗(yàn)和侵襲實(shí)驗(yàn)檢測HMGB1敲低后對(duì)T24細(xì)胞增殖、周期、遷移以及侵襲能力的影響;使用共聚焦顯微鏡和透射電鏡觀察LC3熒光斑點(diǎn);Western blot法檢測相關(guān)蛋白的表達(dá)。結(jié)果:HMGB1在癌組織中的表達(dá)顯著高于癌旁組織(P0.05);CCK8結(jié)果提示,相較于Blank組和NC組,HMGB1干擾組細(xì)胞增殖受抑制;流式細(xì)胞術(shù)提示敲低干擾組中G0/G1期細(xì)胞數(shù)量增加,且吉西他濱誘導(dǎo)的細(xì)胞凋亡增加;劃痕測試法和Transwell侵襲試驗(yàn)顯示敲低HMGB1后細(xì)胞遷移以及侵襲能力減弱;Western blot結(jié)果顯示敲低HMGB1后E-cadherin蛋白表達(dá)上調(diào),N-cadherin、vimentin、MMP-2、MMP-9、cyclinD1、c-Myc、β-catenin蛋白表達(dá)下調(diào),同時(shí)可進(jìn)一步增加吉西他濱誘導(dǎo)的caspase 3和PARP1的活化。結(jié)論:HMGB1可通過促進(jìn)膀胱癌細(xì)胞EMT進(jìn)而增強(qiáng)其惡性生物學(xué)行為,也可通過促進(jìn)自噬活化而參與化療耐藥的過程,使膀胱癌細(xì)胞對(duì)藥物敏感性降低。
[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.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R737.14

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相關(guān)期刊論文 前2條

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