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目的:探讨p16基因和RASSF1A基因甲基化与肺癌发生发展的关系和应用于诊断的意义。方法:采用甲基化特异性PCR(Methylation Specific PCR,MSP)检测120例周边型非小细胞肺癌患者癌组织、痰液脱落细胞和120例非肺癌人群的痰液脱落细胞中p16基因和RASSF1A基因甲基化,分析它们与临床特征的关系以及非肺癌人群与肿瘤患者之间的差异。结果:(1)120例周边型非小细胞肺癌组织中,p16基因甲基化率46.7%(56例),RASSF1A基因甲基化率53.3%(64例)。P16和RASSF1A基因甲基化与吸烟程度、肿瘤大小和临床分期正相关(P<0.05)。(2)肺癌痰液脱落细胞中有28例p16基因出现甲基化(23.3%),20例RASSF1A基因出现甲基化(16.7%),其中32例至少存在一个基因的甲基化(26.7%);66例重度吸烟者中只有4例痰液脱落细胞出现p16基因甲基化(6%),4例出现RASSF1A基因甲基化(6%);54例非重度吸烟正常人中仅有2例出现p16基因甲基化(3.7%),RASSF1A基因无甲基化。(3)液基痰细胞病理学检查与痰脱落细胞p16和RASSF1a基因甲基化检测结合起来可有效提高诊断的灵敏度(P<0.05)。结论:烟草可能具有潜在的诱导抑癌基因p16和RASSF1A发生甲基化的作用;p16和RASSF1A基因甲基化可能参与肺癌的生长过程。痰脱落细胞p16和RASSF1a基因甲基化检测结合液基痰细胞病理学诊断,可提高非小细胞肺癌诊断的灵敏度。
Objective: To investigate the relationship between the methylation of p16 gene and RASSF1A gene and the occurrence and development of lung cancer and its significance for diagnosis. Methods: Methylation-specific PCR (MSP) was used to detect the p16 gene and RASSF1A gene in sputum exfoliated cells in 120 cases of non-small cell lung cancer patients with cancerous tissue, sputum exfoliated cells and 120 non-lung cancer patients Methylation, analyze their relationship with clinical features and the differences between non-lung cancer and cancer patients. Results: (1) The methylation rate of p16 gene was 46.7% (56 cases) and RASSF1A gene methylation rate was 53.3% (64 cases) in 120 cases of peripheral non-small cell lung cancer. Methylation of P16 and RASSF1A genes was positively correlated with the degree of smoking, tumor size and clinical stage (P <0.05). (2) There were 28 cases of p16 gene methylation (23.3%) and 20 cases of RASSF1A gene methylation (16.7%) in lung cancer sputum exfoliated cells, of which 32 cases had at least one gene methylation (26.7% ). Only 4 out of 66 severe smokers had methylation of p16 gene (6%) in sputa exfoliated cells and methylation of RASSF1A gene (6%) in 4 cases. Only 54 out of 54 non-severe smoking smokers Cases showed p16 gene methylation (3.7%), RASSF1A gene without methylation. (3) The combination of liquid-based sputum cytopathology examination and methylation detection of p16 and RASSF1a gene in sputum exfoliated cells can improve the diagnostic sensitivity (P <0.05). Conclusion: Tobacco may potentially induce the methylation of the tumor suppressor genes p16 and RASSF1A. Methylation of p16 and RASSF1A genes may be involved in the growth of lung cancer. Sputum exfoliated cells p16 and RASSF1a gene methylation detection combined with liquid-based sputum cytopathology diagnosis, can improve the sensitivity of non-small cell lung cancer diagnosis.