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为了探讨原癌基因ras和肿瘤抑制基因P53在肝癌中的变化,我们利用免疫组化技术检测了福尔马林固定、石蜡包埋肝癌组织内ras P21、P53蛋白的过量表达,利用聚合酶链式反应-单链构象多态性分析(PCR-SSCP)技术分析了ras和P53基因部分外显子的单链构象多态性。 在25例活检手术外检标本中,免疫组化检测结果5例P21阳性,8例P53阳性,其中有3例为ras P21、P53同时阳性。对P21、P53免疫组化阳性的标本利用PCR-SSCP分析检测ras 12、13和59、61密码子,P53第5、6、7、8外显子的基因突变,发现3例在ras 59、61密码子有异常电泳带,4例在P53第7外显子,2例在第5外显子、第6外显子、第8外显子各1例有异常泳动带,其中有1例在第7、8外显子上均发现异常电泳带。实验表明:1.P53基因突变多发生在分化差的肿瘤中。2.在肝癌的发生发展过程中,同时有原癌基因ras的激活和肿瘤抑制基因P53的灭活。
In order to investigate the changes of proto-oncogene ras and tumor suppressor gene P53 in hepatocellular carcinoma, we used immunohistochemistry to detect over-expression of ras P21 and P53 protein in formalin-fixed, paraffin-embedded hepatocellular carcinoma, using polymerase chain. Single-stranded conformational polymorphisms of partial exons of ras and P53 genes were analyzed by the reaction-single strand conformation polymorphism analysis (PCR-SSCP) technique. Among 25 specimens of biopsy specimens, immunohistochemical results showed that 5 cases were P21 positive and 8 cases were P53 positive, of which 3 cases were ras P21 and P53 positive simultaneously. For the samples with positive P21 and P53 immunohistochemistry, PCR-SSCP analysis was used to detect the ras 12, 13 and 59, 61 codons, and mutations in the 5th, 6th, 7th, and 8th exon of P53 were detected. Three cases were found in ras 59, There were abnormal electrophoresis bands in 61 codons, 4 exons in P53, 2 exons in exon 5, 6 exons, and 1 exon in 8 patients. Abnormal electrophoresis bands were found on exon 7 and exon 8 in each case. Experiments show that: 1. P53 mutations occur in poorly differentiated tumors. 2. During the development of liver cancer, both the activation of the proto-oncogene ras and the inactivation of the tumor suppressor gene P53 were simultaneously observed.