摘要
目的 了解抑癌基因PTEN/MMAC1/TEP1(以下简称PTEN)在肺癌中的缺失和失活。方法 选用 2 4例具有正常对照的肺癌新鲜标本 ,18例小细胞肺癌石蜡切片 ,用聚合酶链反应及杂合性丢失分析法检测PTEN的杂合性丢失 ;并采用原位杂交法、免疫组织化学染色法、Westernblot法观察肺癌标本中PTENmRNA和蛋白水平的表达。对 1株肺腺癌和 3株小细胞肺癌细胞系进行PTEN的Southern、Northern和Westernblot分析。结果 有正常配对的肺癌新鲜标本 45 8% ( 11/ 2 4)有PTEN位点的杂合性丢失 ;mRNA和蛋白表达缺失有 2 5 % ( 6 / 2 4) ,原位杂交与免疫组织化学染色和 (或 )Westernblot结果具有一致性。免疫组织化学染色显示PTEN蛋白表达缺失在小细胞肺癌中占 44 % ( 8/ 18)。 4株细胞系中 1株小细胞肺癌有DNA的纯合性丢失以及RNA和蛋白的阴性表达。结论 肺癌中存在PTENDNA水平的杂合性丢失和RNA。
Objective The recently identified PTEN/MMAC1/TEP1 gene is the first tumor suppressor encoding a phosphatase,which mutated in multiple tumors. Our research aims to detect loss of heterozygosity and inactivation of PTEN in lung cancer. Methods 24 lung cancer fresh samples paired with normal tissue, 18 archival paraffinembedded sections of small cell lung cancer (SCLC) were included. For 24 fresh samples, PCRLOH was used to detect Loss of heterozygosity (LOH) in or near PTEN loci. In situ hybridization, Western blot and or immunohistochemistry were proceeded for mRNA and protein expression of PTEN in lung cancer samples. Southern, Northern and Western blot were done to exam PTEN abnormality in one NSCLC and three SCLC cell lines. Results Among these 24 fresh samples PTEN LOH was detected in 11 out of 24 cases(45.8%) and 6 cases had no PTEN mRNA and protein expression. Results in situ hybridization were in conformity with that of immunohistochemistry/Western blot in our research. No protein expression was detected in 8/18(44%) archival paraffin cases of SCLC. Southern,Northern and Western blot confirmed homozygous deletion in one SCLC cell line. Conclusion Lung cancers had LOH and inactivation of PTEN.
出处
《中华病理学杂志》
CAS
CSCD
北大核心
2000年第2期85-88,共4页
Chinese Journal of Pathology
