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目的 研究肺癌组织中p16和RB基因失活的比率和失活的机制 ,探讨其与肺癌生物学特性及临床、病理和基因分型诊断的关系。方法 采用免疫组化、双重原位杂交、PCR、PCR SSCP以及序列分析等方法 ,检测 10 6例肺癌患者的癌组织和正常肺组织以及 2 3例肺良性疾病标本抑癌基因p16和RB的改变 ,并做对比研究。结果 肺癌细胞p16、RB在蛋白和mRNA水平的表达明显低于正常肺组织和良性疾病肺组织 ,且与肺癌组织学类型、有无淋巴结转移以及临床病理分期密切相关。较早期肺癌 (Ⅰ、Ⅱ期 )病例即有较显著的抑癌基因p16和RB的失活 (32 .6 %和 2 8.3% ) ;非小细胞肺癌以p16基因失活为主 (5 0 .1% ) ,小细胞肺癌以RB基因失活为主 (88.2 % )。p16基因失活的主要机制有纯合缺失、甲基化和点突变。结论 抑癌基因p16和RB在肺癌发生发展中起重要作用 ;p16和RB基因的失活 ,有可能是肺癌发生的早期始动环节 ,对肺癌的早期诊断有重要意义 ;在此基础上 ,有可能建立肺癌基因分型诊断新模式。
Objective To study the ratio of inactivation of p16 and RB genes in lung cancer tissues and the mechanism of inactivation, and to explore the relationship between lung cancer biological characteristics and clinical, pathological and genotypic diagnosis. Methods Immunohistochemistry, double in situ hybridization, PCR, PCR, SSCP, and sequence analysis were used to detect the changes of tumor suppressor gene p16 and RB in 106 lung cancer patients and normal lung tissues and in 23 lung benign diseases. , and do a comparative study. Results The expressions of p16 and RB in lung cancer cells at the protein and mRNA levels were significantly lower than those in normal lung tissues and benign lung tissues, and were closely related to histological type, lymph node metastasis, and clinical pathological stage of lung cancer. In the early stage of lung cancer (stages I and II), there were more significant tumor suppressor gene p16 and RB inactivation (32.6% and 28.3%), and non-small cell lung cancer was mainly inactivated by p16 gene (50. 1%), small cell lung cancer with RB gene inactivation (88.2%). The main mechanisms of p16 gene inactivation are homozygous deletion, methylation and point mutation. Conclusion The tumor suppressor genes p16 and RB play an important role in the development of lung cancer. The inactivation of p16 and RB genes may be the early initiation of lung cancer. It is important for the early diagnosis of lung cancer. On this basis, there are It is possible to establish a new model for the diagnosis of lung cancer genotyping.