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Promoter hypermethylation of p14 ARF, RB, and INK4 gene family in hepatocellular carcinoma with hepatitis B virus infection

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Tumor Biology

Abstract

Both hepatitis B virus (HBV) and gene methylation play important roles in hepatocarcinogenesis. However, their association between HBV infection and gene methylation is not fully understood. Cell cycle control involving RB1 gene-related cell inhibitors is one of the main regulatory pathways were reported to be altered in hepatocellular carcinoma (HCC). The purpose of this research is to assess the methylation status of p14 ARF and INK4 gene family (p14 ARF, p15 INK4B, p16 INK4A, and p18 INK4C) in HCC with HBV infection and HCC without it, and discuss possible role of HBV-induced hypermethylation in the mechanism of hepatocarcinogenesis. Methylation status of RB, p14 ARF, and INK4 gene family in 64 case of HCC with HBV infection and 24 cases without it were detected by methylation-specific polymerase chain reaction, and HBV-DNA of the plasma were detected by quantitative real-time polymerase chain reaction. p14 ARF, p15 INK4B, p16 INK4A, and RB hypermethylation were observed in 30 (34.1 %), 50 (56.8 %), 62 (70.5 %), and 24(27.3 %) of 88 hepatocellular carcinomas, respectively. Methylation frequencies of them between HCC with HBV infection and HCC without it were 43.8 % versus 8.3 % (p14 ARF), 68.9 % versus 25 % (p15 INK4B), 90.6 % versus 16.7 % ( p16 INK4A), and 28.1 % versus 25 % (RB), respectively. In HBV-associated HCC, the numbers of methylated genes were also more than HCC without virus infection, more than two methylated genes were seen in 48 of 64 (75 %) cases; more than three methylated genes were found in 32 of 64 (50 %); correspondently, no one case has more than two genes methylated. p18 INK4C methylation product was not found in cancerous or non-cancerous tissues of 88 HCC. HBV infection is associated with p14 ARF, p15 INK4B, p16 INK4A, and RB gene methylation (P = 0.048, 0.035, 0.02); HBV-DNA replication is associated with p14 ARF, p15 INK4B, p16 INK4A, and RB gene methylation (P = 0.048, 0.035, 0.02); high rate of p14 ARF, p15 INK4B, and p16 INK4A in HCC with HBV infection suggests that HBV-induced hypermethylation may be one of the mechanisms of HBV involved in hepatocellular carcinogenesis.

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Acknowledgments

This work funded by Research Project of Department of Education of Hubei Province (Z20082401).

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Authors and Affiliations

  1. Department of Laboratory Medicine, Taihe Hospital Affiliated to Hubei University of Medicine, Shiyan, 442000, People’s Republic of China

    Ji-Cai Zhang, Bo Gao, Zong-Tao Yu, Jun Lu & Fei Xie

  2. Department of Gastroenterology, Taihe Hospital Affiliated to Hubei University of Medicine, Shiyan, 442000, People’s Republic of China

    Xiao-Bo Liu

  3. Guiyang Medical College, Guiyang, 550004, People’s Republic of China

    Hai-Jun Luo

  4. GCP Office, Taihe Hospital Affiliated to Hubei University of Medicine, No. 32, Reminnan Road, Shiyan, 442000, People’s Republic of China

    Hai-Ping Li

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  1. Ji-Cai Zhang
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Correspondence to Hai-Ping Li.

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Zhang, JC., Gao, B., Yu, ZT. et al. Promoter hypermethylation of p14 ARF, RB, and INK4 gene family in hepatocellular carcinoma with hepatitis B virus infection. Tumor Biol. 35, 2795–2802 (2014). https://doi.org/10.1007/s13277-013-1372-0

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  • DOI: https://doi.org/10.1007/s13277-013-1372-0

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