热损伤对肝细胞肝癌细胞增殖及侵袭转移特性影响的实验研究

发布时间：2018-03-04 00:14

本文选题：热损伤　切入点：微波消融　出处：《介入放射学杂志》2017年05期 　论文类型：期刊论文

【摘要】：目的体外实验评价热损伤对肝细胞肝癌(HCC)细胞增殖、侵袭转移能力及上皮-间质细胞转化(EMT)等特性的影响,探索热消融与HCC复发转移之间的关系。方法通过体外加热构建Mc ARH7777 HCC细胞热损伤模型。CCK-8法检测热损伤对HCC细胞增殖能力的影响,流式细胞技术检测细胞周期情况。Transwell实验研究热损伤对HCC细胞侵袭能力的影响。荧光定量-聚合酶链反应(RTPCR)和蛋白印迹(Western blot)分析热损伤对HCC细胞侵袭及EMT相关分子标志物VEGF、MMP-9、Nm23、E-cadherin、vimentin的m RNA和蛋白表达的影响。结果 Mc A-RH7777 HCC细胞热处理条件为43.5℃水浴30 min。热处理后2~5 d细胞增殖能力显著高于对照组(P0.05)。热处理48 h及72 h后处于G1期细胞比例降低,S+G2期细胞比例增加,差异有统计学意义(P0.05)。与对照组比较,热处理后24 h后HCC细胞侵袭能力差异不明显,而热处理72 h后细胞侵袭能力显著增加(22.3±2.46对14.2±1.82,P0.001)。RT-PCR和蛋白印迹分析结果显示,热处理72 h后HCC细胞VEGF、MMP-9和vimentin表达水平显著增加,E-cadherin表达显著降低,差异有统计学意义(P0.05)。结论亚致死量热损伤诱导Mc ARH7777 HCC细胞发生EMT并增加其增殖和侵袭转移能力,表现出更高的恶性潜能。
[Abstract]:Objective to evaluate the in vitro thermal damage of hepatocellular carcinoma (HCC) cell proliferation, invasion and metastasis of epithelial mesenchymal transition (EMT) of the relationship between HCC and explore the thermal ablation of recurrence and metastasis. Methods constructed by in vitro detection method Mc ARH7777.CCK-8 thermal heating HCC cell damage influence on the thermal damage model HCC cell proliferation, flow cytometry to detect the cell cycle of.Transwell experimental study on the effect of thermal injury on the invasion ability of HCC cells. Fluorescence quantitative polymerase chain reaction (RTPCR) and Western blotting (Western blot) thermal damage analysis on HCC cell invasion and EMT related molecular markers VEGF, MMP-9, Nm23, E-cadherin. The expression of M RNA and vimentin protein. Results 2~5 D cell proliferation in 43.5 DEG C water bath 30 min. after heat treatment was significantly higher than the control group for the Mc A-RH7777 HCC cells (P0.05) heat treatment conditions of heat. 48 h and 72 h of cells in G1 phase decreased, the proportion of cells in S+G2 phase increased, the difference was statistically significant (P0.05). Compared with the control group, after heat treatment at 24 h after the invasion ability of HCC cells was not significantly different, and the heat treatment of the cell invasion ability increased significantly after 72 h (22.3 + 2.46 to 14.2 + 1.82, P0.001).RT-PCR and Western blot analysis showed that the heat treatment of HCC VEGF cells after 72 h, the expression level of MMP-9 and vimentin increased significantly, the expression of E-cadherin was decreased, the difference was statistically significant (P0.05). Conclusion sublethal heat injury induced by Mc ARH7777 HCC EMT cells and increased their proliferation and invasion and metastasis that showed higher malignant potential.

【作者单位】：复旦大学附属中山医院介入科、上海市影像医学研究所;
【基金】：国家自然科学基金面上项目(81171432) 上海市自然科学基金(15ZR1406700) 上海市卫生计生委计划项目(201440540)
【分类号】：R735.7

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