摘要
目的探讨INK4位点反义非编码RNA(ANRIL)下调对CDKN2B-CDKN2A-ARF基因簇的调控及对人肺癌细胞生长的影响。方法通过RNA干扰下调人肺鳞状细胞癌细胞系H520中ANRIL的表达,实时荧光定量聚合酶链反应检测RNA干扰前后细胞中ANRIL mRNA的表达变化,绘制细胞生长曲线;实时荧光定量聚合酶链反应和蛋白质印迹法检测RNA干扰前后细胞中细胞同期蛋白依赖性激酶抑制剂2B(CDKN2B)、CDKN2A、ARF mRNA和蛋白的表达变化。结果 RNA干扰可显著下调ANRIL mRNA的表达(P<0.05),ANRIL下调能抑制人肺鳞状细胞癌细胞系H520的生长(P<0.05),引起CDKN2B、CDKN2A、ARF mRNA和蛋白的表达升高(P<0.05)。结论 RNA干扰AN-RIL能抑制人肺鳞状细胞癌细胞的生长,ANRIL可能通过调控CDKN2B-CDKN2A-ARF基因簇起作用。
Objective To investigate the regulation of CDKN2B-CDKN2A-ARF gene cluster through down-regulation the antisense non-coding RNA in the INK4 locus（ANRIL）,and its influence on the growth of lung cancer H520 cell line.Methods Expression of ANRIL was knocked down by RNA interference in lung squamous cell carcinoma H520 cell line.mRNA expression of ANRIL was demonstrated by real-time polymerase chain reaction（RT-PCR）,and cell growth curve was made.Protein and mRNA expression of cyclin-dependent kinase inhibitor 2B（CDKN2B）,CDKN2A,ARF gene cluster were demonstrated by Western blot and RT-PCR,respectively.Results RNA interference decreased ANRIL mRNA expression（P〈0.05）,and the decreased ANRIL expression in H520 cell line caused reduction of cell proliferation（P〈0.05）,and increased expression of CDKN2B,CDKN2A,ARF gene cluster（P〈0.05）.Conclusion The RNA interference with ANRIL can inhibit the cell growth of human lung squamous cell cancer,and the ANRIL may work through the regulation of CDKN2B-CDKN2A-ARF gene cluster.
出处
《新乡医学院学报》
CAS
2013年第4期259-261,共3页
Journal of Xinxiang Medical University
