注射用核糖核酸Ⅱ对人横纹肌肉瘤细胞增殖和凋亡的调控作用  

Regulation of ribonucleic acid Ⅱ onproliferation and apoptosis of rhabdomyosarcomas RD cells

作　　者：单宇 白雪莲 杨淑贤[1] 韩嘉媛[1] 赵青舟 包晓威 李立勇[1] 曹丽[1] SHAN Yu;BAI Xue-lian;YANG Shu-xian;HAN Jia-yuan;ZHAO Qing-zhou;BAO Xiao-wei;LI Li-yong;CAO Li(Research Center for Pharmacology and Toxicology,Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100193,China;Research Center of Life Sciences and Environment Sciences,Harbin University of Commerce,Harbin 150076,China)

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所药理毒理中心,北京100193 [2]哈尔滨商业大学生命科学与环境科学研究中心,黑龙江哈尔滨150076

出　　处：《中国药理学通报》2018年第11期1505-1511,共7页Chinese Pharmacological Bulletin

基　　金：国家科技部"重大新药创制"科技重大专项(No 2011ZX0920110113)

摘　　要：目的研究注射用核糖核酸Ⅱ调控人恶性胚胎横纹肌瘤RD细胞增殖和凋亡作用的分子机制。方法注射用核糖核酸Ⅱ与RD细胞作用24 h后,MTT法检测细胞存活率,确定给药浓度;Hoechst 33258、AO/EB染色法检测细胞凋亡;JC-1法检测线粒体膜电位;流式细胞术检测细胞周期分布;Western blot检测细胞增殖与凋亡相关蛋白表达。结果注射用核糖核酸Ⅱ可呈浓度依赖性降低RD细胞存活率,诱导细胞凋亡,并明显降低线粒体膜电位。流式细胞术结果表明,RD细胞复制被阻滞在S期。Western blot结果表明,给药后周期阻滞蛋白p21表达上调,CDK2、CDK4下调,p-Akt、p-JNK下调;凋亡相关因子Bcl-2表达下调,Bax上调,caspase-8、caspase-3含量降低。结论注射用核糖核酸Ⅱ通过激活p21,下调G_1/S期关键蛋白CDK2、CDK4,将RD细胞周期阻滞在S期,抑制Akt、JNK的磷酸化,从而抑制RD细胞增殖;通过下调Bcl-2,上调Bax,破坏线粒体完整性,激活caspase-8、caspase-3,诱导RD细胞凋亡。Aim To explore the mechanism of ribonucleic acid II regulating proliferation and apoptosis of embryonal rhabdomyosarcomas RD cells.Methods The RD cells were treated with ribonucleic acid II and cell viability was detected by MTT assay.After ribonucleic acid II treatment for 24 hours,Hoechst 33258 staining and AO/EB staining were adopted for cell apoptosis observation.The mitochondrion membrane potential was measured by JC-1 assay.Cell cycle distribution was detected by flow cytometry.Western blot was chosen to analyze the expressions of related regulators.Results After treatment with ribonucleic acidⅡ,cell viability decreased obviously in a concentration-dependent manner,RD cell apoptosis increased,and the mitochondrion membrane potential decreased markedly.Flow cytometry results manifested ribonucleic acid II blocked cell cycle in phase S.Western blot results showed the decrease of expression of p-Akt,p-JNK.The increase of cell cycle arrest-related protein p21 was accompanied by the reductions of CDK2,CDK4.Apoptosis-related protein Bcl-2 was down-regulated,Bax was up-regulated,and caspase-8 and caspase-3 decreased.Conclusions Ribonucleic acidⅡactivates p21,down-regulates the expression of CDK2 and CDK4,the key factors of phase G 1/S.The phosphorylation levels of Akt and JNK decreased.All in all,it could inhibit the proliferation of RD cells and weaken tumor-formation ability.As well,ribonucleic acidⅡregulates the expressions of Bcl-2 and Bax,destroying the integrity of the mitochondria.Afterwards,caspase-8 and caspase-3 outflow and are activated,promoting the apoptosis of RD cells.

关 键 词：注射用核糖核酸Ⅱ 人横纹肌肉瘤 RD细胞 细胞周期 细胞凋亡 JNK 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]