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目的通过测定3例非小细胞肺癌(NSCLC)细胞株中7个抑癌基因启动子甲基化与mRNA转录之间的关系,探讨启动子甲基化在NSCLC发生中的作用。方法应用甲基化特异性的多聚酶链反应(MSP)和RT-PCR分别检测甲基化状态和mRNA的转录水平。结果肺癌细胞株A549发生甲基化的基因包括p16INK4a、RASSF1α、CDH1、MGMT和CDH13,而DAPK和RAR-β为去甲基化状态;SH-77发生甲基化的基因包括p16INK4a、RASSF1α、CDH1和MGMT,而DAPK、CDH13和RAR-β为去甲基化状态;SPC-A1发生甲基化的基因包括p16INK4a、CDH1、CDH13和RAR-β,而RASSF1α、MGMT和DAPK为去甲基化状态。RASSF1α、MGMT和RAR-β等3个基因发生甲基化的细胞株其mRNA转录失活,而去甲基化状态的细胞株则存在转录活性。结论NSCLC细胞株中抑癌基因经常发生启动子CpG岛的甲基化,并且甲基化可能与抑癌基因mRNA转录失活相关。
Objective To investigate the relationship between promoter methylation and mRNA transcription of 7 tumor suppressor genes in 3 non-small cell lung cancer (NSCLC) cell lines and to explore the role of promoter methylation in the genesis of NSCLC. Methods Methylation-specific polymerase chain reaction (MSP) and RT-PCR were used to detect methylation status and mRNA transcription level respectively. Results The methylation of lung cancer cell line A549 included p16INK4a, RASSF1α, CDH1, MGMT and CDH13, while DAPK and RAR-β were demethylated. The genes involved in SH-77 methylation included p16INK4a, RASSF1α, CDH1 And MGMT, while DAPK, CDH13 and RAR-β are demethylated. The genes methylated by SPC-A1 include p16INK4a, CDH1, CDH13 and RAR-β, while RASSF1α, MGMT and DAPK are demethylated . RASSF1α, MGMT and RAR-β and other 3 genes methylated cell line mRNA inactivation, while the demethylation state of the cell line there is transcriptional activity. Conclusion CpG island methylation of tumor suppressor gene often occurs in NSCLC cell lines, and methylation may be related to the transcriptional inactivation of tumor suppressor gene.