摘要
目的研究miR-223对Lewis肺癌细胞(LLC)周期调控作用及其分子机制。方法将miR-223真核表达载体转染至LLC细胞中,应用流式细胞仪检测其细胞周期分布变化,实时定量PCR、western blot检测对CDK2表达和ERK信号通路的影响。结果流式细胞周期检测显示miR-223过表达导致G0/G1细胞由53.8%减少到45.3%,G2细胞由4.29%增加到13.2%;实时定量PCR及western blot显示,miR-223过表达抑制CDK2 mRNA及蛋白表达,并下调ERK信号通路。结论 miR-223通过作用CDK2及ERK通路参与Lewis肺癌细胞周期调控。
Objective To explore the functional impact of miR-223 on cell cycle regulation of Lewis lung carcinoma（LLC） and to research into its molecular mechanism.Methods Eukaryotic expression vector of miR-223 was transfected in LLC cells.Flowcytometry was applied to detect the changes in the cell cycle of LLC and realtime-RT-PCR and western blot were used to investigate the effects of miR-223 on the expression of CDK2 and ERK signaling.Results The results of flowcytometry showed that the proportion of G0/G1 phase cell in miR-223 expressed group was reduced from 53.8% to 45.3% while the proportion of G2 phase cell in miR-223 was increased from 4.29% to 13.2%.The results from realtime-RT-PCR and western blot indicated that the expressions of CDK2 mRNA in miR-223 expressed group were reduced and ERK signaling was significantly inhibited.Conclusion miR-223 was extensively involved in cell cycle regulation of LLC by targeting CDK2 expression and ERK signaling.
出处
《局解手术学杂志》
2011年第3期231-233,236,共4页
Journal of Regional Anatomy and Operative Surgery
基金
国家高技术研究发展计划"863"项目(2007AA02Z129)
国家自然科学基金项目(30901790)
重庆市自然科学基金项目(2008BB5117)
