半乳糖配体介导Cmyc反义锁核酸对肝癌细胞增殖和凋亡的影响.docVIP

半乳糖配体介导Cmyc反义锁核酸对肝癌细胞增殖和凋亡的影响 　　【摘要】目的探?针对增殖相关基因（Cmyc）第二外显子翻译起始区的反义锁核酸对肝癌细胞增殖和凋亡的影响。 　　方法设计合成能特异性封闭Cmyc基因mRNA第二外显子翻译起始区的反义寡核苷酸、硫代寡核苷酸和锁核酸，分别以阳离子半乳糖配体介导转染HepG2细胞，采用RTPCR法检测细胞内Cmyc的mRNA表达变化；Western blot法检测细胞内Cmyc的蛋白表达变化；流式细胞技术检测细胞凋亡情况；MTT法检测反义锁核酸的细胞毒性作用。 　　结果转染第5 d后，反义锁核酸组Cmyc mRNA相对表达量为（0.335±0.016），明显低于对照组（1.014±0.022），差异有统计学意义（P=0.015）；Cmyc蛋白相对表达量为（0.448±0.037），也明显低于对照组（1.00±0.00），差异有统计学意义（P=0.008）；细胞凋亡比例为（32±6）%，显著高于对照组（0），差异有统计学意义（P=0.032）。 　　结论针对Cmyc第二外显子翻译起始区的反义锁核酸能有效抑制肝癌细胞增殖和促进细胞凋亡。 　　【关键词】肝细胞癌；Cmyc基因；反义锁核酸；外显子2 　　中图分类号：R961文献标识码：ADOI：10.3969/j.issn2017.01.001 　　Effects of Cmyc locked nucleic acid mediated by galactose ligand on proliferation and apoptosis of hepatoma carcinoma cell 　　[HJ1*2][HJ] 　　XIAO Shurong，DENG Yibin▲，XU Guidan，HUANG Zansong 　　（Center for Medical Laboratory Science， Affiliated Hospital of Youjiang Medical University for Nationalities， Baise 533000， China） 　　[HJ2][HJ] 　　【Abstract】ObjectiveTo observe effect of proliferationassociated gene Myc locked nucleic acid（LNA） on the second exon of translation initiation region on proliferation and apoptosis of hepatoma carcinoma cell. 　　MethodsAntisense oligonucleotides， phosphorothioate oligonucleotides and nucleic acids on the second exon of translation initiation region which could specifically block Cmyc mRNA were designed and synthesized. And transfection HepG2 cells were mediated by cationic galactose ligand， mRNA expression in Cmyc was detected by RTPCR， expression of Cmyc protein in cells was detected by Western blot method， cell apoptosis was detected by flow cytometry， and cytotoxicity of locked nucleic acid was detected by MTT， respectively. 　　Results5 days after transfection， relative expression of Cmyc mRNA levels in LNA group was （0.335±0.016）， significantly lower than that （1.014±0.022） of control group， difference was statistically significant（P=0015）. Relative expression quantity of Cmyc protein of the LNA group was （0.448±0.037）， also significantly lower tha