肺癌p14ARF和p16INK4a基因协同表达缺失及其意义

Simultaneous loss of p14ARF and p16INK4a expression in lung cancer cells and its significance

目的 :研究抑癌基因位点INK4a ARF在肺肿瘤细胞中的表达状况 ,揭示p14ARF和p16INK4a协同表达缺失与肺癌发生发展的相关性。方法 :用RT PCR和Westernblot对 6株肺癌细胞 (SPC A 1,Calu 1,H44 6 ,SH77,A5 49,H46 0 )的INK4a ARF基因位点在mRNA、蛋白质水平上进行检测 ,对PCR产物进行纯化和测序分析。结果 :6株肺癌细胞中 ,有 3株细胞 (H46 0 ,Calu 1,A5 49)的p16INK4a基因表达缺失 ,其中 ,Calu 1表达正常p14ARF的mRNA ,而在H46 0和H5 49这两株非小细胞肺癌中 ,p14ARF和p16INK4a发生协同表达缺失。结论 :p14ARF基因与p16INK4a一样 ,是肺肿瘤细胞中的失活靶点之一。揭示了p14ARF缺失导致p5

Purpose:To study the status of the tumor supperessor gene locus INK4a ARF in human lung cancer cells and assess its relationship with the carcinogenesis of lung cancer.Methods:With RT PCR and Western blot,we examined the INK4a ARF gene expression status of 6 lung cancer cell lines (SPC A 1,Calu 1,H446,SH77,A549,H460),on both the mRNA and the protein levels,and the PCR products were sequenced and analyzed.Results:p16INK4a mRNA was undetectable in 3 of these cell lines (H460,Calu 1,A549),while p14ARF was simultaneously missing in H460 and A549,two adenocarcinomas,but still expressed in Calu 1, a squamous cell carcinoma.The p14ARF protein of H460 and A549 were also negative with the Western blot analysis. Conclusions:Like p16INK4a,p14ARF gene is also one of the targets of inactivation in human lung cancer. The finding is a good evidence for the relationship between carcinogenesis and the loss of p14ARF p53 pathway which is considered as an important mechanism of self protection in cells.