Journal of Gastroenterology

, Volume 54, Issue 5, pp 427–436 | Cite as

Independent and additive effects of PNPLA3 and TM6SF2 polymorphisms on the development of non-B, non-C hepatocellular carcinoma

  • Maneerat Raksayot
  • Natthaya Chuaypen
  • Apichaya Khlaiphuengsin
  • Nutcha Pinjaroen
  • Sombat Treeprasertsuk
  • Yong Poovorawan
  • Yasuhito Tanaka
  • Pisit TangkijvanichEmail author
Original Article—Liver, Pancreas, and Biliary Tract



This study was aimed at evaluating the association between single nucleotide polymorphisms (SNPs) in the PNPLA3, NCAN, TM6SF2 and MBOAT7 and hepatocellular carcinoma (HCC) development in Thai patients according to underlying etiologies of liver disease.


These SNPs were determined by allelic discrimination in blood samples of 105 healthy controls and 530 patients with HCC [270 with hepatitis B virus (HBV-HCC), 131 with hepatitis C virus (HCV-HCC), and 129 with non-B, non-C HCC (NBNC-HCC) matched for age and gender].


G allele of PNPLA3 rs738409 variant was significantly higher in NBNC-HCC (49%) compared to healthy controls (32%), HBV-HCC (32%) and HCV-HCC (31%) (P < 0.001). T allele of TM6SF2 rs58542926 was more prevalent in NBNC-HCC (24%) than in healthy controls (8%), HBV-HCC (10%) and HCV-HCC (12%) (P < 0.001). The distribution of NCAN (rs2228603) and MBOAT7 (rs641738) was not different between groups. In multivariate logistic regression analysis, PNPLA3 rs738409 (OR 2.06, 95% CI 1.24–3.43; P = 0.005) and TM6SF2 rs58542926 (OR 2.22, 95% CI 1.34–3.65; P = 0.002) were independently associated with NBNC-HCC compared to viral-related HCC (VR-HCC). The proportion of patients with NBNC-HCC increased significantly along with the increase of the number of risk alleles. There was no association between these SNPs and overall survival in patients with HCC.


These data showed that PNPLA3 and TM6SF2 polymorphisms were independently linked to NBNC-HCC but not HBV- or HCV-HCC in Thai populations. In addition, the risk genotypes might interact with each other through tumor development in patients with NBNC-HCC.


Polymorphisms PNPLA3 TM6SF2 Hepatocellular carcinoma Viral hepatitis 



This study was funded by the Grant for Chula Research Scholar (CU-GRS-60-06-30-03) and the Rachadapisek Sompot Fund for Postdoctoral Fellowship, Chulalongkorn University. This study was also supported by the Thailand Research Fund (RTA5980008), the Research Chair Grant from the National Science and Technology Development Agency (P-15-50004) and JSPS KAKENHI Grant Number JP15H05289. The work was also supported by the Center of Excellence in Hepatitis and Liver Cancer and the Center of Excellence in Clinical Virology, Faculty of Medicine, Chulalongkorn University.


  1. 1.
    Forner A, Llovet JM, Bruix J. Hepatocellular carcinoma. Lancet. 2012;379:1245–55.CrossRefGoogle Scholar
  2. 2.
    El-Serag HB. Epidemiology of viral hepatitis and hepatocellular carcinoma. Gastroenterology. 2012;142(1264–73):e1.Google Scholar
  3. 3.
    Tangkijvanich P, Hirsch P, Theamboonlers A, et al. Association of hepatitis viruses with hepatocellular carcinoma in Thailand. J Gastroenterol. 1999;34:227–33.CrossRefGoogle Scholar
  4. 4.
    Day CP. From fat to inflammation. Gastroenterology. 2006;130:207–10.CrossRefGoogle Scholar
  5. 5.
    Fan JG, Kim SU, Wong VW. New trends on obesity and NAFLD in Asia. J Hepatol. 2017;67:862–73.CrossRefGoogle Scholar
  6. 6.
    Nahon P, Zucman-Rossi J. Single nucleotide polymorphisms and risk of hepatocellular carcinoma in cirrhosis. J Hepatol. 2012;57:663–74.CrossRefGoogle Scholar
  7. 7.
    Romeo S, Kozlitina J, Xing C, et al. Genetic variation in PNPLA3 confers susceptibility to nonalcoholic fatty liver disease. Nat Genet. 2008;40:1461–5.CrossRefGoogle Scholar
  8. 8.
    Anstee QM, Seth D, Day CP. Genetic factors that affect risk of alcoholic and nonalcoholic fatty liver disease. Gastroenterology. 2016;150(1728–44):e7.Google Scholar
  9. 9.
    Speliotes EK, Yerges-Armstrong LM, Wu J, et al. Genome-wide association analysis identifies variants associated with nonalcoholic fatty liver disease that have distinct effects on metabolic traits. PLoS Genet. 2011;7:e1001324.CrossRefGoogle Scholar
  10. 10.
    Kozlitina J, Smagris E, Stender S, et al. Exome-wide association study identifies a TM6SF2 variant that confers susceptibility to nonalcoholic fatty liver disease. Nat Genet. 2014;46:352–6.CrossRefGoogle Scholar
  11. 11.
    Buch S, Stickel F, Trepo E, et al. A genome-wide association study confirms PNPLA3 and identifies TM6SF2 and MBOAT7 as risk loci for alcohol-related cirrhosis. Nat Genet. 2015;47:1443–8.CrossRefGoogle Scholar
  12. 12.
    Mancina RM, Dongiovanni P, Petta S, et al. The MBOAT7-TMC4 variant rs641738 increases risk of nonalcoholic fatty liver disease in individuals of european descent. Gastroenterology. 2016;150(1219–30):e6.Google Scholar
  13. 13.
    Bruix J, Sherman M. Management of hepatocellular carcinoma: an update. Hepatology. 2011;53:1020–2.CrossRefGoogle Scholar
  14. 14.
    Llovet JM, Di Bisceglie AM, Bruix J, et al. Design and endpoints of clinical trials in hepatocellular carcinoma. J Natl Cancer Inst. 2008;100:698–711.CrossRefGoogle Scholar
  15. 15.
    Sopipong W, Tangkijvanich P, Payungporn S, et al. The KIF1B (rs17401966) single nucleotide polymorphism is not associated with the development of HBV-related hepatocellular carcinoma in Thai patients. Asian Pac J Cancer Prev. 2013;14:2865–9.CrossRefGoogle Scholar
  16. 16.
    Nault JC. Pathogenesis of hepatocellular carcinoma according to aetiology. Best Pract Res Clin Gastroenterol. 2014;28:937–47.CrossRefGoogle Scholar
  17. 17.
    Jenkins CM, Mancuso DJ, Yan W, et al. Identification, cloning, expression, and purification of three novel human calcium-independent phospholipase A2 family members possessing triacylglycerol lipase and acylglycerol transacylase activities. J Biol Chem. 2004;279:48968–75.CrossRefGoogle Scholar
  18. 18.
    Pingitore P, Pirazzi C, Mancina RM, et al. Recombinant PNPLA3 protein shows triglyceride hydrolase activity and its I148 M mutation results in loss of function. Biochim Biophys Acta. 2014;1841:574–80.CrossRefGoogle Scholar
  19. 19.
    Bruschi FV, Claudel T, Tardelli M, et al. The PNPLA3 I148 M variant modulates the fibrogenic phenotype of human hepatic stellate cells. Hepatology. 2017;65:1875–90.CrossRefGoogle Scholar
  20. 20.
    Smagris E, BasuRay S, Li J, et al. Pnpla3I148 M knockin mice accumulate PNPLA3 on lipid droplets and develop hepatic steatosis. Hepatology. 2015;61:108–18.CrossRefGoogle Scholar
  21. 21.
    Trepo E, Romeo S, Zucman-Rossi J, et al. PNPLA3 gene in liver diseases. J Hepatol. 2016;65:399–412.CrossRefGoogle Scholar
  22. 22.
    Singal AG, Manjunath H, Yopp AC, et al. The effect of PNPLA3 on fibrosis progression and development of hepatocellular carcinoma: a meta-analysis. Am J Gastroenterol. 2014;109:325–34.CrossRefGoogle Scholar
  23. 23.
    Trepo E, Nahon P, Bontempi G, et al. Association between the PNPLA3 (rs738409 C > G) variant and hepatocellular carcinoma: evidence from a meta-analysis of individual participant data. Hepatology. 2014;59:2170–7.CrossRefGoogle Scholar
  24. 24.
    Seko Y, Sumida Y, Tanaka S, et al. Development of hepatocellular carcinoma in Japanese patients with biopsy-proven non-alcoholic fatty liver disease: association between PNPLA3 genotype and hepatocarcinogenesis/fibrosis progression. Hepatol Res. 2017;47:1083–92.CrossRefGoogle Scholar
  25. 25.
    Ueyama M, Nishida N, Korenaga M, et al. The impact of PNPLA3 and JAZF1 on hepatocellular carcinoma in non-viral hepatitis patients with type 2 diabetes mellitus. J Gastroenterol. 2016;51:370–9.CrossRefGoogle Scholar
  26. 26.
    Brouwer WP, van der Meer AJ, Boonstra A, et al. The impact of PNPLA3 (rs738409 C > G) polymorphisms on liver histology and long-term clinical outcome in chronic hepatitis B patients. Liver Int. 2015;35:438–47.CrossRefGoogle Scholar
  27. 27.
    Chan AW, Wong GL, Chan HY, et al. Concurrent fatty liver increases risk of hepatocellular carcinoma among patients with chronic hepatitis B. J Gastroenterol Hepatol. 2017;32:667–76.CrossRefGoogle Scholar
  28. 28.
    Gao X, Liu W, Yang L, et al. Association between PNPLA3 gene polymorphisms and risk of hepatitis B virus-related hepatocellular carcinoma in Han population in China: a case-control study. Scand J Gastroenterol. 2017;52:1120–7.CrossRefGoogle Scholar
  29. 29.
    Mahdessian H, Taxiarchis A, Popov S, et al. TM6SF2 is a regulator of liver fat metabolism influencing triglyceride secretion and hepatic lipid droplet content. Proc Natl Acad Sci USA. 2014;111:8913–8.CrossRefGoogle Scholar
  30. 30.
    Pirola CJ, Sookoian S. The dual and opposite role of the TM6SF2-rs58542926 variant in protecting against cardiovascular disease and conferring risk for nonalcoholic fatty liver: a meta-analysis. Hepatology. 2015;62:1742–56.CrossRefGoogle Scholar
  31. 31.
    Falleti E, Cussigh A, Cmet S, et al. PNPLA3 rs738409 and TM6SF2 rs58542926 variants increase the risk of hepatocellular carcinoma in alcoholic cirrhosis. Dig Liver Dis. 2016;48:69–75.CrossRefGoogle Scholar
  32. 32.
    Liu YL, Reeves HL, Burt AD, et al. TM6SF2 rs58542926 influences hepatic fibrosis progression in patients with non-alcoholic fatty liver disease. Nat Commun. 2014;5:4309.CrossRefGoogle Scholar
  33. 33.
    Koo BK, Joo SK, Kim D, et al. Additive effects of PNPLA3 and TM6SF2 on the histological severity of non-alcoholic fatty liver disease. J Gastroenterol Hepatol. 2017;6:1277–85.Google Scholar
  34. 34.
    Wang X, Liu Z, Wang K, et al. Additive effects of the risk alleles of PNPLA3 and TM6SF2 on non-alcoholic fatty liver disease (NAFLD) in a Chinese Population. Front Genet. 2016;7:140.Google Scholar
  35. 35.
    Goossens N, Hoshida Y. Is hepatocellular cancer the same disease in alcoholic and nonalcoholic fatty liver diseases? Gastroenterology. 2016;150:1710–7.CrossRefGoogle Scholar
  36. 36.
    Takeuchi Y, Ikeda F, Moritou Y, et al. The impact of patatin-like phospholipase domain-containing protein 3 polymorphism on hepatocellular carcinoma prognosis. J Gastroenterol. 2013;48:405–12.CrossRefGoogle Scholar
  37. 37.
    Valenti L, Motta BM, Soardo G, et al. PNPLA3 I148M polymorphism, clinical presentation, and survival in patients with hepatocellular carcinoma. PLoS One. 2013;8:e75982.CrossRefGoogle Scholar
  38. 38.
    D’Souza K, Epand RM. Enrichment of phosphatidylinositols with specific acyl chains. Biochim Biophys Acta. 2014;1838:1501–8.CrossRefGoogle Scholar
  39. 39.
    Luukkonen PK, Zhou Y, Hyotylainen T, et al. The MBOAT7 variant rs641738 alters hepatic phosphatidylinositols and increases severity of non-alcoholic fatty liver disease in humans. J Hepatol. 2016;65:1263–5.CrossRefGoogle Scholar
  40. 40.
    Nischalke HD, Lutz P, Kramer B, et al. A common polymorphism in the NCAN gene is associated with hepatocellular carcinoma in alcoholic liver disease. J Hepatol. 2014;61:1073–9.CrossRefGoogle Scholar

Copyright information

© Japanese Society of Gastroenterology 2018

Authors and Affiliations

  • Maneerat Raksayot
    • 1
  • Natthaya Chuaypen
    • 1
  • Apichaya Khlaiphuengsin
    • 1
  • Nutcha Pinjaroen
    • 2
  • Sombat Treeprasertsuk
    • 3
  • Yong Poovorawan
    • 4
  • Yasuhito Tanaka
    • 5
  • Pisit Tangkijvanich
    • 1
    Email author
  1. 1.Center of Excellence in Hepatitis and Liver Cancer, Department of Biochemistry, Faculty of MedicineChulalongkorn UniversityBangkokThailand
  2. 2.Department of Radiology, Faculty of MedicineChulalongkorn UniversityBangkokThailand
  3. 3.Division of Gastroenterology, Department of Medicine, Faculty of MedicineChulalongkorn UniversityBangkokThailand
  4. 4.Center of Excellence in Clinical Virology, Department of Pediatrics, Faculty of MedicineChulalongkorn UniversityBangkokThailand
  5. 5.Department of Virology and Liver UnitNagoya City University Graduate School of Medical SciencesNagoyaJapan

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