We examined whether genetic polymorphisms of efflux transporters in hepatocytes are associated with susceptibility to develop hepatocellular carcinoma (HCC). Genetic polymorphisms of drug transporters expressed in hepatocytes were analyzed using DNA samples from hepatitis C virus (HCV)-seropositive cirrhotic patients with HCC (n = 58), and allele and haplotype frequencies were compared with those in healthy subjects (n = 61). To search for single nucleotide polymorphisms (SNPs) in HCC susceptibility genes, 34 SNPs in 6 efflux transporters [MDR1 (ABCB1), ABCC1, ABCC2, ABCC3, ABCG2 and ABCB11] were determined. No significant association was observed for any single SNP; however, some haplotypes in ABCC1 and ABCB11 were associated with HCC. Furthermore, three combinations of SNPs (3435C > T in ABCB1 and 825T > C in ABCC1), (3435C > T in ABCB1 and -15281_-15278CTCT > delete in ABCB11), and (825T > C in ABCC1 and -15281_-15278CTCT > delete in ABCB11) were significantly associated with HCC. The present study suggests that genetic variations of ABC transporters such as ABCB1, ABCB11, and ABCC1 are associated with susceptibility to develop HCC, implying that aberrant hepatic clearance of toxic substances may increase the risk of hepatocarcinogenesis. Further studies of how these polymor-phisms are associated with phenotypic differences are warranted.
KeywordsGenetic PolymorphismDrug TransporterHepatitis C VirusHepatocellular Carcinoma
H. Tsukuma, T. Hiyama, S. Tanaka, M. Nakao, T. Yabuuchi, T. Kitamura, K. Nakanishi, I. Fujimoto, A. Inoue, H. Yamazaki and T. Kawashima, “Risk Factors for Hepatocellular Carcinoma among Patients with Chronic Liver Disease,” The New England Journal of Medicine, Vol. 328, No. 12, 1993, pp. 17971801.
I. Saito, T. Miyamura, A. Ohbayashi, H. harada, T. Katayama, S. Kikuchi, Y. Watanabe, S. Koi, M. Onji, Y. Ohta, Q.L. Choo, M. Houghton and G. Kuo, “Hepatitis C Virus Infection is Associated with the Development of Hepatocellular Carcinoma,” Proceedings of the National Academy of Sciences of the United States of America, Vol. 87, No. 17, 1990, pp. 65476549.
H. Oka, N. Kurioka, K. Kim, T. Kanno, T. Kuroki, Y. Mizoguchi and K. Kobayashi, “Prospective Study of Early Detection of Hepatocellular Carcinoma in Patients with Cirrhosis,” Hepatology, Vol. 12, No. 4, 1990, pp. 680687.
K. Kiyosawa, T. Umemura, T. Ichijo, A. Matsumoto, K. Yoshizawa, A. Gad and E. Tanaka, “Hepatocellular Carcinoma: Recent Trends in Japan,” Gastroenterology, Vol. 127, No. (Suppl. 1), 2004, pp. S17S26.
E. M. Leslie, R. G. Deeley and S. P. Cole, “Multidrug Resistance Proteins: Role of PGlycoprotein, MRP1, MRP2, and BCRP (ABCG2) in Tissue Defense,” Toxicology and Applied Pharmacology, Vol. 204, No. 3, 2005, pp. 216237.
L. R. Ferguson and S. de Flora, “Multiple Drug Resistance, Antimutagenesis and Anticarcinogenesis,” Mutation Research, Vol. 591, No. 12, 2005, pp. 2433.
G. C. Yeh, J. Lopaczynska, C. M. Poore and J. M. Phang, “A New Functional Role for PGlycoprotein: Efflux Pump for Benzo(Alpha)Pyrene in Human Breast Cancer MCF7 Cells,” Cancer Research, Vol. 52, No. 23, 1992, pp. 66926695.
I. Ieiri, H. Takane, T. Hirota, K. Otsubo and S. Higuchi, “Genetic Polymorphisms of Drug Transporters: Pharmacokinetic and Pharmacodynamic Consequences in Pharmacotherapy,” Expert Opinion on Drug Metabolism & Toxicology, Vol. 2, No. 5, 2006, pp. 651674.
S. Sookoian, G. Castano, A. Burgueno, T. F. Gianotti and C. J. Pirola, “Association of the MultidrugResistance Associated Protein Gene (ABCC2) Variants with Intrahepatic Cholestasis of Pregnancy,” Journal of Hepatology, Vol. 48, No. 1, 2008, pp. 125132.
L. I. Hu , X. X. Wang, X. Chen, J. Chang, C. Li, Y. Zhang, J. Yang, W. Jiang and S. M. Zhuang, “BCRP Gene Polymorphisms are Associated with Susceptibility and Survival of Diffuse Large BCell Lymphoma,” Carcinogenesis, Vol. 28, No. 8, 2007, pp. 17401744.
Y. Korenaga, K. Naito, N. Okayama, H. Hirata, Y. Suehiro, Y. Hamanaka, H. Matsuyama and Y. Hinoda, “Association of the BCRP C421A Polymorphism with Nonpapillary Renal Cell Carcinoma,” International Journal of Cancer, Vol. 117, No. 3, 2005, pp. 431434.
M. Tanabe, I. Ieiri, N. Nagata, K. Inoue, S. Ito, Y. Kanamori, M. Takahashi, Y. Kurata, J. Kigawa, S. Hoguchi, N. Terakawa and K. Otsubo, “Expression of PGly coprotein in Human Placenta: Relation to Genetic Polymorphism of the Multidrug Resistance (MDR)1 Gene,” Journal of Pharmacology and Experimental Therapeutics, Vol. 297, No. 3, 2001, pp. 11371143.
G. D. Leschziner, T. Andrew, M. Pirmohamed and M. R. Johnson, “ABCB1 Genotype and PGP Expression, Function and Therapeutic Drug Response: A Critical Review and Recommendations for Future Research,” The Pharmacogenomics Journal, Vol. 7, No. 3, 2007, pp. 154179.
I. Ieiri, H. Takane and K. Otsubo, “The MDR1 (ABCB1) Gene Polymorphism and its Clinical Implications,” Clinical Pharmacokinetics, Vol. 43, No. 9, 2004, pp. 553 576.
S. Ito, I. Ieiri, M. Tanabe, A. Suzuki, S. Higuchi and K. Otsubo, “Polymorphism of the ABC Transporter Genes, MDR1, MRP1 and MRP2/cMOAT, in Healthy Japanese Subjects,” Pharmacogenetics, Vol. 11, No. 2, 2001, pp. 175184.
H. FukushimaUesaka, Y. Saito, K. Maekawa, R. Hasegawa, K. Suzuki, T. Yanagawa, H. Kajio, N. Kuzuya, M. Noda, K. Yasuda, M. Tohkin and J. Sawada, “Genetic Variations of the ABC Transporter Gene ABCC3 in a Japanese Population,” Drug Metabolism and Pharmaco kinetics, Vol. 22, No. 2, 2007, pp. 129135.
D. Kobayashi, I. Ieiri, T. Hirota, H. Takane, S. Maegawa, J. Kigawa, H. Suzuki, E. Nanba, M. Oshimura, N. Terakawa, K. Otsubo, K. Mine and Y. Sugiyama, “Functional Assessment of ABCG2 (BCRP) Gene Polymorphisms to Protein Expression in Human Placenta,” Drug Metabolism and Disposition, Vol. 33, No. 1, 2005, pp. 94101.
T. Lang, M. Haberl, D. Jung, A. Drescher, R. Schlagen haufer, A. Keil, E. Mornhinweg, B. Stieger, G. A. Kullak Ublick and R. Kerb, “Genetic Variability, Haplo type Structures, and Ethnic Diversity of Hepatic Trans porters MDR3 (ABCB4) and Bile Salt Export Pump (ABCB11),” Drug Metabolism and Disposition, Vol. 34, No. 9, 2006, pp. 15821599.
T. Gerloff, B. Stieger, B. Hagenbuch, J. Madon, L. Landmann, J. Roth, A. F. Hoffmann and P. J. Meier, “The Sister of PGlycoprotein Represents the Canalicular Bile Salt Export Pump Of Mammalian Liver,” The Journal of Biological Chemistry, Vol. 273, No. 16, 1998, pp. 10046 10050.
R. Wang, M. Salem, I. M. Yousef, B. Tuchweber, P. Lam, S. J. Childs, C. D. Helgason, C. Ackerley, M. J. Phillips and V. Ling, “Targeted Inactivation of Sister of PGlyco protein Gene (spgp) in Mice Results in Nonprogressive but Persistent Intrahepatic Cholestasis,” Proceedings of the National Academy of Sciences of the United States of America, Vol. 98, No. 4, 2001, pp. 20112016.
S. S. Strautnieks, L. N. Bull, A. S. Knisely, S. A. Kocoshis, N. Dahl, H. H. Arnell, E. Sokal, K. Dahan, S. Childs, V. Ling, M. S. Tanner, A. F. Kagalwalla, A. Nemeth, J. Pawlowska, A. Baker, G. MieliVergani, N. B. Freimer, R. M. Gardiner and R. J. Thompson, “A Gene Encoding a LiverSpecific ABC Transporter is Mutated in Progressive Familial Intrahepatic Cholestasis,” Nature Genetics, Vol. 20, No. 3, 1998, pp. 233238.
P. L. Jansen, S. S. Strautnieks and E. Jacquemin, “Hepa tocanalicular Bile Salt Export Pump Deficiency in Patients with Progressive Familial Intrahepatic Cholestasis,” Gastroenterology, Vol. 117, No. 6, 1999, pp. 13701379.
E. M. Leslie, K. Ito, P. Upadhyaya, S. S. Hecht, R. G. Deeley and S. P. Cole, “Transport of the BetaOGlu curonide Conjugate of the TobaccoSpecific Carcinogen 4(Methylnitrosamino)1(3Pyridyl)1Butanol (NNAL) by the Multidrug Resistance Protein 1 (MRP1). Re quirement for Glutathione or a NonSulfurContaining Analog,” The Journal of Biological Chemistry, Vol. 276, No. 30, 2001, pp. 2784627854.
A. E. van Herwaarden, J. W. Jonker, E. Wagenaar, R. F. Brinkhuis, J. H. M. Schellens, J. H. Beijnen and A. H. Schinkel, “The Breast Cancer Resistance Protein (Bcrp1/ Abcg2) Restricts Exposure to the Dietary Carcinogen 2Amino1Methyl6Phenylimidazo[4,5b]Pyridine,” Cancer Research, Vol. 63, No. 19, 2003, pp. 64476452.
M. Siegsmund, U. Brinkmann, E. Scháffeler, G. Weirich, M. Schwab, M. Eichelbaum, P. Fritz, O. Burk, J. Decker, P. Alken, U. Rothenoieler, R. Kerb, S. Hoffmeyer and S. Brauch, “Association of the PGlycoprotein Transporter MDR1(C3435T) Polymorphism with the Susceptibility to Renal Epithelial Tumors,” Journal of the American Society of Nephrology, Vol. 13, No. 7, 2002, pp. 18471854.
U. Potocnik, D. Glavac and M. Dean, “Common Germline MDR1/ABCB1 Functional Polymorphisms and Haplotypes Modify Susceptibility to Colorectal Cancers with High Microsatellite Instability,” Cancer Genetics and Cytogenetics, Vol. 183, No. 1, 2008, pp. 2834.
T. E. Massey, R. K. Stewart, J. M. Daniels and L. Liu, “Biochemical and Molecular Aspects of Mammalian Susceptibility to Aflatoxin B1 Carcinogenicity,” Procee dings of the Society for Experimental Biology and Medicine, Vol. 208, No. 3, 1995, pp. 213227.
D. W. Loe, R. K. Stewart, T. E. Massey, R. G. Deeley and S. P. Cole, “ATPDependent Transport of Aflatoxin B1 and its Glutathione Conjugates by the Product of the Multidrug Resistance Protein (MRP) Gene,” Molecular Pharmacology, Vol. 51, No. 6, 1997, pp. 10341041.
A. Lorico, J. Nesland, E. Emilsen, O. Fodstad and G. Rappa, “Role of the Multidrug Resistance Protein 1 Gene in the Carcinogenicity of Aflatoxin B1: Investigations Using MRP1Null Mice,” Toxicology, Vol. 171, No. 2, 2002, pp. 201205.
E. Ros, L. Libbrecht, M. Geuken, P. L. Jansen and T. A. Roskams, “High Expression of MDR1, MRP1, and MRP3 in the Hepatic Progenitor Cell Compartment and Hepatocytes in Severe Human Liver Disease,” Journal of Pathology, Vol. 200, No. 5, 2003, pp. 553560.
S. Bonin, L. Pascolo, L. S. Crocé, G. Stanta and C. Tiribelli, “Gene Expression of ABC Proteins in Hepatocellular Carcinoma, Perineoplastic Tissue, and Liver Diseases,” Molecular Medicine, Vol. 8, No. 4, 2002, pp. 318325.
G. A. Meijer, A. B. Schroeijers, M. J. Flens, S. G. Meu wissen, P. van der Valk, J. P. A. Baak and R. J. Scheper, “Increased Expression of Multidrug Resistance Related Proteins PGP, MRP1, and LRP/MVP Occurs Early in Colorectal Carcinogenesis,” Journal of Clinical Pathology, Vol. 52, No. 6, 1999, pp. 450454.
A. S. Knisely, S. S. Strautnieks, Y. Meier, B. Stieger, J. A. Byne, B. C. Portmann, L. N. Bull, L. Pawlikowska, B. Bilezik?i, F. ?z?ay, A. László, L. Tiszlavicz, L. Moore, J. Raftos, H. Arnell, B. Fischer, A. Németh, N. Papadogiannakis, J. CieleckaKuszyk, I. Jankowska, J. Pawlowska, H. MelínAldana, K. M. Emerick, P. F. Whitington, G. Mieli Vergani and R. J. Thompson, “Hepatocellular Carcinoma in Ten Children under Five Years of Age with Bile Salt Export Pump Deficiency,” Hepatology, Vol. 44, No. 2, 2006,
H. Bernstein, C. Bernstein, C. M. Payne, K. Dvorakova and H. Garewal, “Bile Acids as Carcinogens in Human Gastrointestinal Cancers,” Mutation Research, Vol. 589, No. 1, 2005, pp. 4765.
S. Hoffmeyer, O. Burk, O. von Richter, H. P. Arnold, J. Brockm?ller, A. Johne, I. Cascorbi, T. Gerloff, I. Roots, M. Eichelbaum and U. Brinkmann, “Functional Polymorphisms of the Human MultidrugResistance Gene: Multiple Sequence Variations and Correlation of One Allele with PGlycoprotein Expression and Activity in vivo,” Proceedings of the National Academy of Sciences of the United States of America, Vol. 97, No. 7, 2000, pp. 34733478.
D. Wang and W. Sadée, “Searching for Polymorphisms that Affect Gene Expression and mRNA Processing: Example ABCB1 (MDR1),” The AAPS Journal, Vol. 8, No. 3, 2006, pp. E515E520.
C. KimchiSarfaty, J. M. Oh, I. W. Kim, Z. E. Sauna, A. M. Calcagno, S. V. Ambudkar and M. M. Gottesman, “A ‘Silent’ Polymorphism in the MDR1 Gene Changes Substrate Specificity,” Science, Vol. 315, No. 5811, 2007, pp. 525528.