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N-ras gene mutation and hepatitis B virus infection in hepatocellular carcinomas in Guangxi, China*
http://www.100md.com 《世界华人消化杂志》 1998年第6期
hepatocellularcarcinoma|N-rasgene|HBV|AFB1,N-rasgenemutationandhepatitisBvirusinfectioninhepatocellularcarcinomasinGuangxi,China*,关键词:
     LIU Qi-Fu1 , LUO Dan1 , SU Jian-Jia1 , C Gove2 and R Williams2 世界华人消化杂志 1998 0 0 6


    关键词:hepatocellular carcinoma; N-ras gene; HBV; AFB1 期刊 sjhrxhzz 0 Original Articles fur -->


    

Abstract

    AIM To observe the roles of N-ras gene mutation and hepatitis B virus (HBV) infection in the carcinogenesis of hepatocellular carcinoma (HCC) in Guangxi, China.

    METHODS The polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and immunohistochemistry were used to detect N-ras gene mutation and HBV infection in 29 cases of HCC.

    RESULTS The aberration rates at codon 2-37 of N-ras were 79.31% in HCCs and 80.77% in adjacent non-tumorous liver tissues. More than 2 point mutations of N-ras gene were observed in 22 (75.86%) cases. HBsAg and HBxAg positive rates were 86.2% and 79.3%. There was a parallel tendency between HBV marker detections and the mutation rate of N-ras gene.

    CONCLUSION HBV infection and N-ras gene mutation may be involved in the carcinogenesis and development of HCC in Guangxi. Since the aflatoxin B1 contamination is one of risk factors for HCC in this area, it may contribute to the mutation of N-ras gene in carcinogenesis of HCC.INTRODUCTION

    Hepatocellular carcinoma (HCC) is one of common malignant tumors in People′s Republic of China. Guangxi is a high incidence area of HCC. Many factors are involved in hepatocarcinogenesis. Many studies revealed that hepatitis B virus (HBV) infection might be a risk factor for hepatocellular carcinogenesis. One theory for hepatocarcinogenesis is that the oncogene(s) may be transactivated by hepatitis B x antigen (HBxAg)[1] . It is found recently that activation of N-ras gene may be the molecular basis for the carcinogenesis and development of HCC[2,3] . There have been reports about overexpression of N-ras oncogene in human HCC[4] , but a few dealt with the roles of N-ras gene mutation and HBV infection, and their relationship with HCC. We analyzed the N-ras gene mutation and HBV infection in HCC by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and immunohistochemistry in 29 cases of human HCC.

    MATERIALS AND METHODS

    Clinical samples

    Surgically resected specimens of HCC (29 cases) were collected in Guangxi Cancer Institute during the period of 1987-1992. Twenty-eight cases of them contained HCC adjacent liver tissues. All samples were fixed with 10% formalin, embedded in paraffin and stained with haematoxylin and eosin (HE).

    Immunohistochemistry

    Immunostaining was performed by a streptavidin-biotin immunoperoxidase method. Hepatitis B surface antigen (HBsAg) and x antigen (HBxAg) were detected with monoclonal anti-HBsAg antibody, anti-HBxAg antibody and Strept ABC kit (DAKO A/S Denmark).

    DNA extraction

    Genomic DNA was prepared by the proteinase K-Phenol-Chloroform extraction method.

    PCR-SSCP

    Oligomers that flank codon 2-37 of N-ras genes were synthesized as primers by the Department of Molecular Medicine, King′s College Hospital, UK. One of them was 5′-end labeled with r-32 P ATP by T4 polynucleotide kinase reaction. The primer sets were as follows:

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