人肝细胞癌c-myc基因扩增、MTS1/p16基因变异和HBV感染的研究

Correlation of c-myc gene amplification, MTS1/p16 gene alternation, and HBV infection in human hepatocellular carcinoma

目的 探讨原癌基因c-myc扩增、抑癌基因MTS1/p16变异以及HBV感染在人肝细胞癌（HCC）发生发展中的作用。 方法 应用差异PCR结合非变性聚丙烯酰胺凝胶电泳银染及激光扫描（d-PCR-PAGE-Laser）技术检测c-myc基因扩增，应用PCR结合单链构象多态（SSCP）银染法分析MTS1/p16基因变异，PCR检测HBV DNA。 结果 (1)29例配对肝细胞癌、癌旁组织中c-myc基因扩增阳性率分别为44.83%（13/29）和51.72%（15/29），两者差异无显著性，P＞0.05；但两者均显著高于肝硬化组织c-myc基因扩增的阳性率8.33%（1/12），P＜0.05。(2)只有3例（10.34%，3/29）肝细胞癌中发现MTS1/p16基因纯合性缺失，未发现MTS1/p16基因突变。(3)正常肝脏、肝硬化和肝细胞癌组织中HBV DNA的阳性率分别为14.29%（2/14）、66.67%（8/12）和96.55%（28/29），三者间差异具有高度显著性，P＜0.001，并且HBV DNA的阳性率随肝脏病情的加重而增高（b＝0.3986, P＜0.001）。(4)29例肝细胞癌中c-myc基因扩增和HBV DNA存在与否无关（P＜0.01）。 结论 (1)c-myc基因扩增和HBV感染与HCC的发生、发展密切相关，HCC中c-myc基因扩增和HBV感染之间无内在相关性。(2)HCC中MTS1/p16基因纯合性缺失和突变的发生频率较低。

Objective To observe the roles of c-myc gene amplification, MTS1/p16 gene alternation, and HBV infection in the pathogenesis and progression of hepatocellular carcinoma (HCC). Methods A d-PCR-PAGE-laser scanning technique was used to define amplifications of c-myc oncogene. Alternations of MTS1/p16 gene exon1 and exon2 were analysed by PCR and single-strand conformation polymorphism (SSCP) silver stain method. HBV-DNA was assayed by PCR. Results (1)The positive rates of c-myc gene amplification of HCCs and their paired non-cancerous liver tissues were 44.83% (13/29) and 51.72% (15/29). The discrepancies between them were not significant (P>0.05), but they were both significantly higher than that of cirrhotic liver tissues (8.33%, 1/12, P<0.05). (2)A total of 3 homozygous deletions and no mutations of MTS1/p16 gene exon1 and exon2 in HCCs were found in the subset of HCCs. (3)The discrepancies of the positive rates of HBV-DNA among normal liver (14.29%, 2/14), cirrhotic liver (66.67%, 8/12) and HCCs (96.55%, 28/29) were significant (P<0.001). Moreover, the HBV-DNA positive rates increased according to the development of liver lesions (b=0.3986, P<0.001). Conclusions (1)C-myc gene amplification and HBV infection are closely related to the development and progression in a subset of HCCs. However, c-myc gene amplification does not correlate with the HBV infection in HCCs. (2)The homozygous deletions and mutations of MTS1/p16 gene are infrequently encountered in the subset of HCCs. [