Effect of Single Nucleotide Polymorphisms (SNPs) on Microrna-221 on Target Gene in Hepatocellular Carcinoma (HCC) Cell Lines

Main Article Content

ธนัญญา จินาโต
ณัฐธยาน์ ช่วยเพ็ญ
พิสิฐ ตั้งกิจวาณิชย์
สัญชัย พยุงภร


Hepatocellular carcinoma (HCC) is a disease with a high mortality rate due to the disease is mostly diagnosed in severe phase. The identification of biomarkers is important for early diagnosis of the disease. MicroRNAs (miRNAs) are small noncoding RNAs that play important roles in regulating the expression of many genes and differentially expressed between patients and healthy persons. Moreover, single-nucleotide polymorphisms (SNPs) in miRNAs are affecting on function of miRNAs. This research focused on the effect of SNPs rs113054794 (A/C) within miR-221 for regulation of Aryl hydrocarbon receptor nuclear translocator (ARNT) gene expression in HCC cell line (HepG2). Validation assay based on construction of mi-221 expression vector then transfected into HepG2 cells and measured the expression level of miR-221 by real-time PCR. The result showed that the expression of miR-221 was 65-folds increased in miR-221 transfected group. In addition, the hybridization pattern and the minimum free energy (mfe) between miR-221 and ARNT gene were analyzed by RNAhybrid web-based software. The result revealed that SNPs (A/C) within miR-221 yielded exactly difference in the hybridization pattern and mfe between rs113054794 and ARNT gene (allele C = -25.9 kcal/mol whereas allele A = -18.7 kcal/mol). After that, the expression level of target gene (ARNT) was measured by real-time PCR in order to test the silencing efficiency of miR-221. The result suggested that both alleles of miR-221 triggered ARNT gene silencing. However, ARNT expression level was non-statistically different between allele C and allele A. Therefore, SNPs (A/C) within miR-221 were affected the regulation of ARNT gene expression. However, these SNPs may regulate other genes, which required further study.


Download data is not yet available.

Article Details

Research Article


H. B. El-Serag, and K. L. Rudolph, “Hepatocellular carcinoma: epidemiology and molecular carcinogenesis,” Gastroenterology, vol. 132, no. 7, Jun, 2007, pp. 2557-76.

K. Hagymasi, and Z. Tulassay, “[Epidemiology, risk factors and molecular pathogenesis of primary liver cancer],” Orv Hetil, vol. 149, no. 12, Mar 23, 2008, pp. 541-8.

V. Bihrer et al., “Serum microRNA-21 as marker for necroinflammation in hepatitis C patients with and without hepatocellular carcinoma,” PLos One, vol. 6, no. 10, 2011, pp. e26971.

D. P. Bartel, “MicroRNAs: genomics, biogenesis, mechanism, and function,” Cell, vol. 116, no. 2, Jan 23, 2004, pp. 281-97.

A. Muniategui et al., “Quantification of miRNA-mRNA interactions,” PLos One, vol. 7, no. 2, 2012, pp. e30766.

K. Uchino et al., “RNAi therapeutics and applications of microRNAs in cancer treatment,” Jpn J Clin Oncol, vol. 43, no. 6, Jun, 2013, pp. 596-607.

S. Khare et al., “Epigenetics of hepatocellular carcinoma: role of microRNA,” World J Gastroenterol, vol. 19, no. 33, Sep 7, 2013, pp. 5439-45.

W. P. Kloosterman, and R. H. Plasterk, “The diverse functions of microRNAs in animal development and disease,” Dev Cell, vol. 11, no. 4, Oct, 2006, pp. 441-50.

J. Li et al., “Expression of serum miR-221 in human hepatocellular carcinoma and its prognostic significance,” Biochem Biophys Res Commun, vol. 406, no. 1, Mar 4, 2011, pp. 70-3.

P. Pineau et al., “miR-221 overexpression contributes to liver

tumorigenesis,” Proc Nati Acad Sci U S A, vol. 107, no. 1, Jan 5, 2010, pp. 264-9.

J. Yang et al., “Prognostic role of microRNA-221 in various human

malignant neoplasms: a meta-analysis of 20 related studies,” PLoS One, vol. 9, no. 1, 2014, pp. e87606.

G. T. Nguyen-Dien et al., “Genetic polymorphisms in miRNAs targeting the estrogen receptor and their effect on breast cancer risk,” Meta Gene, vol. 2, no. 0, 12//, 2014, pp. 226-236.

J. Brennecke et al., “Principles of microRNA-target recognition,” PLoS Biol, vol. 3, no. 3, Mar, 2005, pp. e85.

Y. Liang et al., “Aryl hydrocarbon receptor nuclear translocator is associated with tumor growth and progression of hepatocellular carcinoma,” Int J Cencer, vol. 130, no. 8, Apr 15, 2012, pp. 1745-54.

Q. Yuan et al., “MicroRNA-221 overexpression accelerates hepatocyte proliferation during liver regeneration,” Hepatology, vol. 57, no. 1, Jan, 2013, pp. 299-310.

Z. Z. Zhang et al., “Hepatitis B virus and hepatocellular carcinoma at the miRNA level,” World J Gastroenterol, vol. 17, no. 28, Jul 28, 2011, pp. 3353-8.

S. O. Yoon et al., “Deregulated expression of microRNA-221 with the potential for prognostic biomarkers in surgically resected hepatocellular carcinoma,” Hum Pathol, vol. 42, no. 10, Oct, 2011, pp. 1391-400.

F. Fornari et al., “MiR-221 controls CDKN1C/p57 and CDKN1B/p27 expression in human hepatocellular carcinoma,” Oncogene, vol. 27, no. 43, Sep 25, 2008, pp. 5651-61.

X. Li et al., “microRNA: a promising diagnostic biomarker and therapeutic target for hepatocellular carcinoma,” Dig Dis Sci, vol. 59, no. 6, Jun, 2014, pp. 1099-107.

M. Rong et al., “Increased miR-221 expression in hepatocellular carcinoma tissues and its role in enhancing cell growth and inhibiting apoptosis in vitro,” BMC Cancer, vol. 13, 2013, pp. 21.