Transactivation of the human MDR1 gene by hepatitis B virus X gene product.
OBJECTIVE
Persistent hepatitis B virus (HBV) infection may cause hepatocellular carcinoma. Patients with hepatocellular carcinoma are characterized by nonresponsiveness to chemotherapeutic agents. While many studies have been devoted to understanding the hepatocarcinogenesis mechanism of HBV, the possible relationship between HBV and the drug sensitivity phenotype of cancer cells has rarely been addressed. The hepatitis B virus X gene encodes a transcription transactivator which has been suggested to be a potential factor in viral hepatocarcinogenesis. The role of HBV pX in mediating the drug resistance phenotype of hepatoma cell lines was examined in this study.


METHODS
Standard transfection and chloramphenicol acetyltransferase assay were utilized to examine the effect of HBV pX transactivator on a reporter gene under the control of the human multidrug resistance (MDR) 1 upstream regulatory elements. Selected Hep G2 clones with or without HBV pX expression were tested for their sensitivity towards various anti-cancer agents by utilization of MTT assay.


RESULTS
The expression of HBV pX in both Hep G2 (p53+) and Hep 3B (p53-) cells resulted in transactivation of the reporter gene under control of the human MDR1 upstream regulatory elements. Northern blot analysis indicated that expression of the endogenous MDR1 gene was also elevated in Hep G2 clones with HBV pX expression. Decreased drug sensitivity towards adriamycin, vinblastine, and VP-16 was observed in Hep G2 clones with HBV pX expression.


CONCLUSIONS
HBV pX can transactivate the MDR1 gene. Drug sensitivity was altered in Hep G2 cells with HBV pX expression.
Doong
S L
SL
Graduate Institute of Microbiology and Cancer Research Center, National Taiwan University, College of Medicine, Taipei, ROC. SLDOONG@HA.MC.NTU.EDU.TW
Lin
M H
MH
Tsai
M M
MM
Li
T R
TR
Chuang
S E
SE
Cheng
A L
AL
D003001:Cloning, Molecular; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D019450:Genes, MDR; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D010641:Phenotype; D013778:Tetrazolium Salts; D014157:Transcription Factors / Q000235:genetics*; D015533:Transcriptional Activation
D013778:Tetrazolium Salts; D014157:Transcription Factors