miRNA153反义核酸上调肺癌细胞对辐射敏感性的体外实验  

Up-regulation of microRNA153 causes radio-sensitization of lung cancer cells in vitro experiments

作　　者：高蕊[1] 冯帆[2] 贾辉[2] 张帆[3] 王涛[4] 董国福[4] 马德宾[1] 马宏达[2] 韩雅玲[1] 刘蕾[1] 

机构地区：[1]沈阳军区总医院呼吸与重症医学科,辽宁沈阳110016 [2]沈阳军区总医院药剂科,辽宁沈阳110016 [3]解放军总医院肿瘤内科,北京100853 [4]军事医学科学院,北京100850

出　　处：《解放军医学院学报》2015年第10期1033-1038,共6页Academic Journal of Chinese PLA Medical School

基　　金："十二五"全军医学科研项目(BWS12J007)~~

摘　　要：目的探讨反义核酸沉默microRNA 153(miRNA153)对电离辐射杀伤肺癌细胞株的影响。方法利用脂质体转染miRNA153的反义核酸;使用60Co-γ射线照射肺癌细胞;使用CCK-8实验、软琼脂成集落实验(锚定非依赖性生长)和Trans-well实验检测mi RNA153反义核酸对电离辐射杀伤肺癌细胞的影响;RT-PCR和Western blot实验检测miRNA153的反义核酸对mi RNA153、其靶标蛋白PTEN以及细胞存活/凋亡耐受调控蛋白Survivin表达的影响。结果 CCK-8实验结果显示,中等剂量射线(4 Gy)照射能够杀伤肺癌细胞A549、H460、H1299和H358,下调mi RNA153的表达能够增强辐射对肺癌细胞的杀伤作用;软琼脂成集落和Transwell实验进一步证实mi RNA153的反义核酸能够上调辐射对A549细胞锚定非依赖性生长和侵袭的抑制作用。分子机制实验结果表明,mi RNA153的反义核酸能够显著降低mi RNA153的表达水平,提高肿瘤抑制因子PTEN并降低细胞存活因子Survivin的表达。结论降低miRNA153的表达能够上调辐射对肺癌细胞系的体外杀伤作用。Objective To declare whether silencing of mi RNA153 via its anti-sense nuclear（inhibitor） induces the radio-sensitization of lung cancer cells. Methods The anti-sense nuclear acid（inhibitor） of mi RNA153 was transfected into lung cancer cells, which were irradiated by 60Co-γ. The CCK-8 analysis, soft-agar and Transwell assays were performed to identify the effect of mi RNA153 silencing on radio-sensitization in lung cancer cells. RT-PCR and Western blot assays were used to detect the effect of anti-sense nuclear acid（inhibitor） of mi RNA153 on mi RNA153, its target protein PTEN and the expression of Survivin cell proliferation of survival regulators. Results CCK-8 analysis revealed that the irradiation of medium dose ray（4 Gy） could kill the lung cancer cells（A549, H460, H1299 and H358）, and the down-regulation of expression of mi RNA153 could enhance the radio-sensitization in lung cancer cells. The soft-agar and Transwell assays proved that the up-regulation of rays could inhibit the anchorage-independent growth and invasion of A549 cells. And the molecular mechanism experiment indicated that the anti-sense nuclear acid of mi RNA153 significantly disrupted the endogenous expression of mi RNA153 and Survivin, and in turn upregulated the expression of PTEN. Conclusion Silencing of mi R-153 significantly enhances the sensitivity of lung cancers.

关 键 词：miRNA153沉默 肺癌细胞 电离辐射 

分 类 号：R73-3[医药卫生—肿瘤]