High-resolution melting analysis reveals genetic polymorphisms in MicroRNAs confer hepatocellular carcinoma risk in Chinese patients

BMC Cancer

Song-Mei Liu³  Xin Zhou³  Subrata Sen²  Jing-Tao Huang³  Fan Shen³  Jinyun Chen¹  Jin Wang²  Jia-Hui Qi³ 

[1]Department of Epidemiology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA

[2]Department of Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas 77054, USA

[3]Center for Gene Diagnosis, Medical Research Center, Zhongnan Hospital of Wuhan University, 169 Donghu Road, Wuhan, Hubei 430071, China

关键词: Single-nucleotide polymorphisms;    High-resolution melting;    MicroRNA;    Hepatocellular carcinoma;   

Others  :  1121150
DOI  :  10.1186/1471-2407-14-643

 received in 2014-06-17, accepted in 2014-08-26,  发布年份 2014

【 摘 要 】

Background

Although several single-nucleotide polymorphisms in microRNA (miRNA) genes have been associated with primary hepatocellular carcinoma, published findings regarding this relationship are inconsistent and inconclusive.

Methods

The high-resolution melting (HRM) analysis was used to determine whether the occurrence of the SNPs of miR-146a C > G (rs2910164), miR-196a2 C > T (rs11614913), miR-301b A > G (rs384262), and miR-499 C > T (rs3746444) differs in frequency-matched 314 HCC patients and 407 controls by age and sex.

Results

The groups’ genotype distributions of miR-196a2 C > T and miR-499 C > T differed significantly (P < 0.01), both of them increased the risk of HCC in different dominant genetic models (P < 0.01); compared with individuals carrying one or neither of the unfavorable genotypes, individuals carrying both unfavorable genotypes (CT + CC) had a 3.11-fold higher HCC risk (95% confidence interval (CI), 1.89–5.09; P = 7.18 × 10⁻⁶). Moreover, the allele frequency of miR-499 C > T was significantly different between the two groups, and the HCC risk of carriers of the C allele was higher than that of carriers of the T allele (odds ratio, 1.53; 95% CI, 1.15-2.03; P = 0.003). Further, we found that the activated partial thromboplastin time (APTT) in HCC patients with miR-196a2 CC genotype was longer than patients with TT genotypes (P < 0.05), and HCC patients with miR-499 C allele had higher serum levels of direct bilirubin, globulin, γ-glutamyltranspeptidase, alkaline phosphatase, and lower serum cholinesterase (P < 0.05).

Conclusions

Our findings suggest that the SNPs in miR-196a2 C > T and miR-499 C > T confer HCC risk and that affect the clinical laboratory characteristics of HCC patients.

【 授权许可】

   
2014 Qi et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

【 参考文献 】

• [1]Fares N, Peron JM: Epidemiology, natural history, and risk factors of hepatocellular carcinoma. Rev Prat 2013, 63(2):216-217. 220–212
• [2]Schwartz M, Roayaie S, Konstadoulakis M: Strategies for the management of hepatocellular carcinoma. Nat Clin Pract Oncol 2007, 4(7):424-432.
• [3]Qi J, Wang J, Katayama H, Sen S, Liu SM: Circulating microRNAs (cmiRNAs) as novel potential biomarkers for hepatocellular carcinoma. Neoplasma 2013, 60(2):135-142.
• [4]El-Serag HB: Epidemiology of viral hepatitis and hepatocellular carcinoma. Gastroenterology 2012, 142(6):e1261-e1273.
• [5]Tanaka M, Katayama F, Kato H, Tanaka H, Wang J, Qiao YL, Inoue M: Hepatitis B and C virus infection and hepatocellular carcinoma in China: a review of epidemiology and control measures. J Epidemiol 2011, 21(6):401-416.
• [6]Bartel DP: MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 2004, 116(2):281-297.
• [7]Wang J, Sen S: MicroRNA functional network in pancreatic cancer: from biology to biomarkers of disease. J Biosci 2011, 36(3):481-491.
• [8]Ambros V: MicroRNA pathways in flies and worms: growth, death, fat, stress, and timing. Cell 2003, 113(6):673-676.
• [9]Esquela-Kerscher A, Slack FJ: Oncomirs - microRNAs with a role in cancer. Nat Rev Cancer 2006, 6(4):259-269.
• [10]Horikawa Y, Wood CG, Yang H, Zhao H, Ye Y, Gu J, Lin J, Habuchi T, Wu X: Single nucleotide polymorphisms of microRNA machinery genes modify the risk of renal cell carcinoma. Clin Cancer Res 2008, 14(23):7956-7962.
• [11]Hu Z, Chen J, Tian T, Zhou X, Gu H, Xu L, Zeng Y, Miao R, Jin G, Ma H, Chen Y, Shen H: Genetic variants of miRNA sequences and non-small cell lung cancer survival. J Clin Invest 2008, 118(7):2600-2608.
• [12]Qi P, Dou TH, Geng L, Zhou FG, Gu X, Wang H, Gao CF: Association of a variant in MIR 196A2 with susceptibility to hepatocellular carcinoma in male Chinese patients with chronic hepatitis B virus infection. Hum Immunol 2010, 71(6):621-626.
• [13]Xiang Y, Fan S, Cao J, Huang S, Zhang LP: Association of the microRNA-499 variants with susceptibility to hepatocellular carcinoma in a Chinese population. Mol Biol Rep 2012, 39(6):7019-7023.
• [14]Xu T, Zhu Y, Wei QK, Yuan Y, Zhou F, Ge YY, Yang JR, Su H, Zhuang SM: A functional polymorphism in the miR-146a gene is associated with the risk for hepatocellular carcinoma. Carcinogenesis 2008, 29(11):2126-2131.
• [15]Xu Y, Li L, Xiang X, Wang H, Cai W, Xie J, Han Y, Bao S, Xie Q: Three common functional polymorphisms in microRNA encoding genes in the susceptibility to hepatocellular carcinoma: A systematic review and meta-analysis. Gene 2013, 527(2):584-593.
• [16]Guo J, Jin M, Zhang M, Chen K: A genetic variant in miR-196a2 increased digestive system cancer risks: a meta-analysis of 15 case–control studies. PLoS One 2012, 7(1):e30585.
• [17]Gao LB, Bai P, Pan XM, Jia J, Li LJ, Liang WB, Tang M, Zhang LS, Wei YG, Zhang L: The association between two polymorphisms in pre-miRNAs and breast cancer risk: a meta-analysis. Breast Cancer Res Treat 2011, 125(2):571-574.
• [18]Hu Z, Liang J, Wang Z, Tian T, Zhou X, Chen J, Miao R, Wang Y, Wang X, Shen H: Common genetic variants in pre-microRNAs were associated with increased risk of breast cancer in Chinese women. Hum Mutat 2009, 30(1):79-84.
• [19]Peng S, Kuang Z, Sheng C, Zhang Y, Xu H, Cheng Q: Association of microRNA-196a-2 gene polymorphism with gastric cancer risk in a Chinese population. Dig Dis Sci 2010, 55(8):2288-2293.
• [20]Kogo R, Mimori K, Tanaka F, Komune S, Mori M: Clinical significance of miR-146a in gastric cancer cases. Clin Cancer Res 2011, 17(13):4277-4284.
• [21]Zhu L, Chu H, Gu D, Ma L, Shi D, Zhong D, Tong N, Zhang Z, Wang M: A functional polymorphism in miRNA-196a2 is associated with colorectal cancer risk in a Chinese population. DNA Cell Biol 2012, 31(3):350-354.
• [22]Vinci S, Gelmini S, Mancini I, Malentacchi F, Pazzagli M, Beltrami C, Pinzani P, Orlando C: Genetic and epigenetic factors in regulation of microRNA in colorectal cancers. Methods 2013, 59(1):138-146.
• [23]Zhou B, Wang K, Wang Y, Xi M, Zhang Z, Song Y, Zhang L: Common genetic polymorphisms in pre-microRNAs and risk of cervical squamous cell carcinoma. Mol Carcinog 2011, 50(7):499-505.
• [24]Shen J, Ambrosone CB, DiCioccio RA, Odunsi K, Lele SB, Zhao H: A functional polymorphism in the miR-146a gene and age of familial breast/ovarian cancer diagnosis. Carcinogenesis 2008, 29(10):1963-1966.
• [25]Jazdzewski K, Murray EL, Franssila K, Jarzab B, Schoenberg DR, de la Chapelle A: Common SNP in pre-miR-146a decreases mature miR expression and predisposes to papillary thyroid carcinoma. Proc Natl Acad Sci U S A 2008, 105(20):7269-7274.
• [26]Permuth-Wey J, Thompson RC, Burton Nabors L, Olson JJ, Browning JE, Madden MH, Ann Chen Y, Egan KM: A functional polymorphism in the pre-miR-146a gene is associated with risk and prognosis in adult glioma. J Neurooncol 2011, 105(3):639-646.
• [27]Chu YH, Tzeng SL, Lin CW, Chien MH, Chen MK, Yang SF: Impacts of microRNA gene polymorphisms on the susceptibility of environmental factors leading to carcinogenesis in oral cancer. PLoS One 2012, 7(6):e39777.
• [28]Wei W, Hu Z, Fu H, Tie Y, Zhang H, Wu Y, Zheng X: MicroRNA-1 and microRNA-499 downregulate the expression of the ets1 proto-oncogene in HepG2 cells. Oncol Rep 2012, 28(2):701-706.
• [29]Zhu K, Pan Q, Zhang X, Kong LQ, Fan J, Dai Z, Wang L, Yang XR, Hu J, Wan JL, Zhao YM, Tao ZH, Chai ZT, Zeng HY, Tang ZY, Sun HC, Zhou J: MiR-146a enhances angiogenic activity of endothelial cells in hepatocellular carcinoma by promoting PDGFRA expression. Carcinogenesis 2013, 34(9):2071-2079.
• [30]Tomokuni A, Eguchi H, Tomimaru Y, Wada H, Kawamoto K, Kobayashi S, Marubashi S, Tanemura M, Nagano H, Mori M, Doki Y: miR-146a suppresses the sensitivity to interferon-alpha in hepatocellular carcinoma cells. Biochem Biophys Res Commun 2011, 414(4):675-680.
• [31]Jiang J, Gusev Y, Aderca I, Mettler TA, Nagorney DM, Brackett DJ, Roberts LR, Schmittgen TD: Association of MicroRNA expression in hepatocellular carcinomas with hepatitis infection, cirrhosis, and patient survival. Clin Cancer Res 2008, 14(2):419-427.
• [32]Zhang X, Daucher M, Armistead D, Russell R, Kottilil S: MicroRNA expression profiling in HCV-infected human hepatoma cells identifies potential anti-viral targets induced by interferon-alpha. PLoS One 2013, 8(2):e55733.
• [33]Hu M, Zhao L, Hu S, Yang J: The association between two common polymorphisms in MicroRNAs and hepatocellular carcinoma risk in Asian population. PLoS One 2013, 8(2):e57012.
• [34]Wittwer CT, Reed GH, Gundry CN, Vandersteen JG, Pryor RJ: High-resolution genotyping by amplicon melting analysis using LCGreen. Clin Chem 2003, 49(6 Pt 1):853-860.
• [35]Vossen RH, Aten E, Roos A, den Dunnen JT: High-resolution melting analysis (HRMA): more than just sequence variant screening. Hum Mutat 2009, 30(6):860-866.
• [36]Lin CW, Er TK, Tsai FJ, Liu TC, Shin PY, Chang JG: Development of a high-resolution melting method for the screening of Wilson disease-related ATP7B gene mutations. Clin Chim Acta 2010, 411(17–18):1223-1231.
• [37]Li SW, Lin K, Ma P, Zhang ZL, Zhou YD, Lu SY, Zhou X, Liu SM: FADS gene polymorphisms confer the risk of coronary artery disease in a Chinese Han population through the altered desaturase activities: based on high-resolution melting analysis. PLoS One 2013, 8(1):e55869.
• [38]Liu SM, Xu FX, Shen F, Xie Y: Rapid genotyping of APOA5–1131 T > C polymorphism using high resolution melting analysis with unlabeled probes. Gene 2012, 498(2):276-279.
• [39]Lin PC, Liu TC, Chang CC, Chen YH, Chang JG: High-resolution melting (HRM) analysis for the detection of single nucleotide polymorphisms in microRNA target sites. Clin Chim Acta 2012, 413(13–14):1092-1097.
• [40]Reed GH, Wittwer CT: Sensitivity and specificity of single-nucleotide polymorphism scanning by high-resolution melting analysis. Clin Chem 2004, 50(10):1748-1754.
• [41]Hosking L, Lumsden S, Lewis K, Yeo A, McCarthy L, Bansal A, Riley J, Purvis I, Xu CF: Detection of genotyping errors by Hardy-Weinberg equilibrium testing. Eur J Hum Genet 2004, 12(5):395-399.
• [42]Kinoshita A, Onoda H, Imai N, Iwaku A, Oishi M, Tanaka K, Fushiya N, Koike K, Nishino H, Matsushima M, Saeki C, Tajiri H: The Glasgow Prognostic Score, an inflammation based prognostic score, predicts survival in patients with hepatocellular carcinoma. BMC Cancer 2013, 13:52. BioMed Central Full Text
• [43]Yu MC, Chan KM, Lee CF, Lee YS, Eldeen FZ, Chou HS, Lee WC, Chen MF: Alkaline phosphatase: does it have a role in predicting hepatocellular carcinoma recurrence? J Gastrointest Surg 2011, 15(8):1440-1449.
• [44]Donadon M, Cimino M, Procopio F, Morenghi E, Montorsi M, Torzilli G: Potential role of cholinesterases to predict short-term outcome after hepatic resection for hepatocellular carcinoma. Updates Surg 2013, 65(1):11-18.
• [45]Sjöholm MI, Hoffmann G, Lindgren S, Dillner J, Carlson J: Comparison of archival plasma and formalin-fixed paraffin-embedded tissue for genotyping inhepatocellular carcinoma. Cancer Epidemiol Biomarkers Prev 2005, 14(1):251-255.