| Hepatocellular carcinoma(HCC) is one of the most common carcinomas in our country. With virus-induced hepatocirrhosis and chronic hepatitis well known as pre-carcinoma disease, HBV has been verified to have closest correlation with HCC. It is often considered that the abnormality of various genetic and epigenetic inheritance mechanisms contributes to the incidence of HCC, but since epigenetic has no influence on the sequence of genes, it's believed that the epigenetic inheritance changes such as DNA methylation can be reversed. Therefore, researchers expect to prevent and treat carcinoma by regulating epigenetic inheritance modification in the future. Furthermore, the promoter methylation of carcinoma-related gene occurred frequently in the development of HCC. 14-3-3 sigma gene is the only known subtype with antitumor activity among all 14-3-3 proteins, which plays a role of negative-regulation in the process of cell cycle and apoptosis, thereby possessing the properties of pathogenetic mechanisms like cell-cycle control, DNA damage reparation, inhibition of apoptosis, promoting cell differentiation and aging and so forth. On the other hand, secreted frizzled-related protein (SFRP) as a negative-regulating protein in Wnt-signaling pathway, SFRP also contributes to the aspects of carcinoma forming as well as apoptosis regulation.Objective:To research the methylation status of 14-3-3 sigma gene,SFRP1 and SFRP2 gene in HBV-related HCC and the relationship between the methylation status of the three genes and the development of HCC.Methods:Using methylation-specific PCR(MSP) to detect methylation status of 14-3-3 sigma gene,SFRP1 and SFRP2 gene of excised 45 specimens of carcinoma and adjacent non-tumorous liver tissues from HCC patients intra-operative, and also excised 6 normal liver tissues from cholecystolithiasis or hemangiomas patients.Then the relationship between the methylation status of every gene and the clinical characteristics was analyzed.Results:After 45 samples amplification by MSP, 14-3-3 sigma gene methylation was detected in 41 HCC tissues and 33 adjacent non-tumorous liver tissues, accounted for 91.1% and 73.3% , SFRP1 gene methylation was detected in 28 HCC tissues and 16 adjacent non-tumorous liver tissues, accounted for 62.2% and 35.6% , and SFRP2 gene methylation was detected in 23 HCC tissues and 13 adjacent non-tumorous liver tissues, accounted for 51.1% and 28.9%.But in 6 samples of normal liver tissues, nonmethylation status were detected of the three genes. There was hypermethylation of the three genes in HCC tissues and adjacent non-tumorous liver tissues. And the statistical correlation of three gnes methylation between HCC and adjacent non-tumorous liver tissues was found(P<0.05). The samples which of the methylation status of three genes in HCC tissues is more than adjacent non-tumorous liver tissues(P<0.05),the difference have a significant statistics. Compared with normal liver tissues, HCC tissues has higher rate of methylation both in 14-3-3σand SFRP1(P<0.05, respectively). However, there is no statistical difference between the methylation rate of SFRP2 gene from HCC tissues and normal liver tissues. Methylation of 14-3-3 sigma gene ,SFRP1 and SFRP2 gene have no correlation with gender, age, serum markers, types of meta-tissue, transferring and patholevel in 45 samples HBV-related HCCtissues (P>0.05). Conclusion: The methylation of 14-3-3 sigma, SFRP1 and SFRP2 is frequently detected in the early stage of HBV-related HCC tissues, which indicates the potential value as a biomarker in predicting HCC. At that time, to detect the methylation of multiple genes can not only provide a new pathway for the diagnosis of HCC, but also may provide a strong theoretical basis for gene therapy of carcinoma in terms of methylation level alteration.
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