Determination of miR-193b rs30236 single nucleotide polymorphism in ovarian cancer patients

Background: Single nucleotide polymorphism (SNP) is the most common form of genetic variability. The miR-193b was connected to several types of tumors and also with occurrence of resistance to chemotherapy. This investigation aimed to determine miR-193b rs30236 polymorphism in ovarian cancer patients and controls. The authors performed bioinformatics analysis to discover the target genes of miR-193b. Materials and Methods: Eighty-six ovarian cancer patients and 102 matched controls were included in the study. Blood samples were drown into EDTA tubes and DNA was isolated by silica adsorption method. Mir-193b rs30236C/T polymorphism was determined by melting curve analysis. Chi-square test was used for statistical evaluation of the data. MiRTargetLink was used for determination of the has-miR-193b-3p microRNA target genes. Results: Based on melting curve analysis of miR-193b rs30236, the authors found 30.24% T allele frequency in the patients’ group, and 35.78% in the control group (p = 0.2549). They detected CC genotype in 39.22%, CT genotype in 50.00%, and TT genotype in 10.78% of controls, while they occurred in 45.35%, 48.84%, and 5.81% of the patients (p = 0.4096), respectively. The most important target genes of miR-193b are cyclin D1 (CCND1), estrogen receptor (ESR1), plasminogen activator (PLAU), prolin rich acidic protein (PRAP1), and myeloid leukemia sequence (MCL1). Conclusions: The miR-193b has effect on the expression of several genes. The rs30236 polymorphism has not been studied in ovarian cancer patients, yet. The authors did not find significant difference in allele and genotype difference in patients and controls. To discover the exact role and suitability of this SNP for use as biomarker in the ovarian cancer prior to chemotherapy, more detailed studies are needed.

miRNA; miR-193b; SNP, Gene expression; Ovarian cancer; Melting curve analysis

[1] Yates L.A., Norbury C.J., Gilbert R.J.: “The long and short of microRNA”. Cell, 2013, 153, 516.

[2] Davidson B., Tropé G.C., Reich R.: “The clinical and diagnostic role of microRNAs in ovarian carcinoma”. Gynecol. Oncol., 2014, 133, 640.

[3] Weng L., Ziliak D., Im H.K., Gamazon E.R., Philips S., Nguyen A.T., et al.: “Genome-wide discovery of genetic variants affecting tamoxifen sensitivity and their clinical and functional validation”. Ann. Oncol., 2013, 24, 1867.

[4] Unno K., Zhou Y., Zimmerman T., Platanias L.C, Wickrema A.: “Identification of a novel microRNA cluster miR-193b-365 in multiple myeloma”. Leuk. Lymphoma, 2009, 50, 1865.

[5] Rauhala H.E., Jalava S.E., Isotalo J., Bracken H., Lehmusvaara S., Tammela T.L., et al.: “miR-193b is an epigenetically regulated putatitive tumor supressor in prostate cancer”. Int. J. Cancer, 2010, 127, 1363.

[6] Lujambio A., Ropero S., Ballestar E., Fraga M.F., Cerrato C., Setién F., et al.: “Genetic unmasking of an epigenetically silenced microRNA in human cancer cells”. Cancer Res., 2007, 67, 1424.

[7] Yong L.F., Law W.C., Wang W.C.: “Potentiality of triple microRNA classifier: miR-193a-3p, miR-23a and miR-339-5p for early detection of colorectal cancer”. BMC Cancer, 2013, 13, 280.

[8] Fang C., Li X.P., Gong W.J., Wu N.Y., Tang J., Yin J.Y., et al.: “Agerelated common miRNA polymorphism associated with severe toxicity in lung cancer patients treated with platinum-based chemotherapy”. Clin. Exp. Pharmacol. Physiol., 2016, 44, 21.

[9] Hu Z., Shu Y., Chen Y., Dong J., Liu Y., Pan S., et al.: “Genetic polymorphism in precursor microRNA flanking region and nonsmall cell lung cancer survival”. Am. J. Respir. Crit. Care Med., 2011, 183, 641.

[10] Leivonen S.K., Makela R., Ostling P., Kohonen P., Haapa-Paananen S., Kleivi K., et al.: “Protein lysate microarray analysis to identify microRNAs regulating estogen receptor signaling in breast cancer cell lines”. Oncogene, 2009, 28, 3926.

[11] Nakamura K., Sawada K., Yoshimura A., Kinose Y., Nakatsuka E., Kimura T.: “Clinical relevance of circulating cell-free microRNAs in ovarian cancer”. Mol. Cancer, 2016 ,15, 48.

[12] Ziliak D., Gamazon R.E., Lacroix B., Kyung I. H., Wen Y., Huang R.S.: “Genetic variation that predicts platinum sensitivity reveals the role of miR-193b in chemotherapeutic susceptibility”. Mol. Cancer Ther., 2012, 11, 2054.

[13] Nagy B., Bán Z., Papp Z.: “The DNA isolation method has effect on allele drop out and on the results of fluorescent PCR and DNA fragment analysis”. Clin. Chim. Acta, 2005, 360, 128.

[14] Huang R.S., Johnatty E.S., Gamazon R.E., Im H.K., Ziliak D., Duan S., et al.: “Platinum sensitivity-related germline polymorphism discovered via a cell-based approach and analysis of its association with outcome in ovarian cancer patients”. Clin. Cancer Res., 2011, 17, 5490.

[15] Torres A., Torres K., Maciejewski R., Harwey M.H.: “MicroRNAs and their role in gynecological tumors”. Med. Res. Rev., 2011, 31, 895.

[16] He L., Thompson J.M., Hemann M.T., Hernando-Monge E, Mu D, Goodson S., et al.: “A microRNA polycistron as a potential human oncogene”. Nature, 2005, 435, 828.

[17] Schetter A.J., Leung S.Y., Sohn J.J., Zanetti K.A., Bowman E.D., Yanaihara N., et al.: “MicroRNA expression profiles associated with prognosis and therapeutic outcome in colon adenocarcinoma”. J. Am. Med. Assoc., 2008, 299, 425.

[18] Johnson S.M., Grosshans H., Shinghara J., Byrom M., Jarvis R., Cheng A., et al.: “RAS is regulated by the let-7 microRNA family. Cell, 2005, 120, 13944.

[19] Cannistra S.A.: “Cancer of the ovary”. N. Engl. J. Med., 2004, 51, 2519.

[20] Dai J., Wei R-J, Li R., Feng J.B., Yu Y.L., Liu P.S.: “A study of CCND1 with epithelial ovarian cancer cell proliferation and apoptosis”. Eur. Rev. Med. Pharmacol. Sci., 2016, 20, 4230.

[21] Zamagni C., Wirtz M..R.., De Iaco P., Rosati M., Veltrup E., Rosati F., et al.: “Oestrogen receptor a mRNA is a prognostic factor in ovarian cancer patients treated with neo-adjuvant chemotherapy: determination by array and kinetic PCR in fresh tissue biopsies. Endocr. Relat. Cancer, 2009, 16, 1241.

[22] Havrilesky J.L., Whitehead M.C., Rubatt M.J., Cheek R.L., Groelke J., He Q., et al.: “Evaluation of biomarker panels for early stage ovarian cancer detection and monitoring for disease recurrence”. Gynecol. Oncol., 2008, 110, 374.

[23] Huang H.B., Zhuo L.J., Leung W.H.C., Lu G.D., Liu J.J., Yap C.T., Hooi S.C.: “PRAP1 is a novel executor of p53-dependent mechanisms in cell survival after DNA damage”. Cell Death Dis., 2012, 3, e442.

[24] Sugio A., Iwasaki M., Habata S.. Mariya T., Suzuki M., Osogami H., et al.: “BAG3 upregulates MCL-1 through downregulation of miR29b to induce anticancer drug resistance in ovarian cancer”. Gynecol. Oncol., 2014, 134, 615.