Association of miR-146a and miR196a2 genotype with susceptibility to idiopathic recurrent pregnancy loss in Iranian women: A case-control study
Background: Recurrent pregnancy loss (RPL) is the most common complaint of pregnancy in females with a prevalence of 5%. Numerous documents have shown that single nucleotide polymorphisms are able to change miRNA transcription and/or maturation, which may alter the incidence of disorders such as RPL.
Objective: To assess the relationship of miR-146aC > G (rs2910164) and miR-196a2T > C (rs11614913) with RPL susceptibility in Iranian women.
Materials and Methods: Blood samples were collected from 214 women who had experienced at least two consecutive spontaneous miscarriages (case) and 147 normal individuals without a history of miscarriage (control). MiR-146aC > G and miR-196a2T > C genotypes were evaluated via the restriction fragment length polymorphism technique.
Results: The genotypes incidence did not show a significant difference in pre-miR-146aC > G polymorphism CC vs CG + GG (p = 0.854; OR = 0.933; 95% CI) and CC + CG vs GG (p = 0.282; OR = 1.454; 95% CI). Also, no significant difference was observed between pre-miR-196a2T > C polymorphism TT vs TC + CC (p = 0.862; OR = 0.938; 95% CI) and TT + TC vs CC and (p = 0.291; OR = 1.462; 95% CI) in both the case and control groups.
Conclusion: The results showed that although the distribution of miR-146aC > G and miR-196a2T > C was different between the unknown RPL and control groups, these variances were not statistically significant.
Key words: RPL, miR-146a, miR196a2, Polymorphism, RFLP.
 Ardestani MT, Nodushan HH, Aflatoonian A, Ghasemi N, Sheikhha MH. Case control study of the factor V Leiden and factor II G20210A mutation frequency in women with recurrent pregnancy loss. Iran J Reprod Med 2013; 11: 61–64.
 Nematollahi Z, Hadinedoushan H, Aflatoonian A, Eslami G, Ghasemi N. The association between single nucleotide polymorphism in interleukin-27 gene and recurrent pregnancy loss in Iranian women. Iran J Reprod Med 2015; 13: 209–214.
 Reddy R, Mutreja D, Mukhopadhyay I, Moorchung N. Prothrombin gene mutations do not cause recurrent pregnancy loss in the Indian population. Int J Reprod Biomed 2018; 17: 375–376.
 Zarezade Z, Ashrafzadeh Mehrjardi HR, Zaimy M, Ghasemi N. Association between rs1264457 A/G polymorphism of HLA-E gene and recurrent pregnancy loss. Iran J Reprod Med 2015; 13 (Suppl.): 72.
 Nabi A, Khalili MA, Halvaei I, Ghasemzadeh J, Zare E. Seminal bacterial contaminations: Probable factor in unexplained recurrent pregnancy loss. Iran J Reprod Med 2013; 11: 925–932.
 Nejatbakhsh F, Bioos S, Sohrabvand F. Simple vaginal medicaments for unexplained infertility in Iranian traditional medicine. Iran J Reprod Med 2013; 2: 63–64.
 Ahmadpour Nazm S, Bonyadi M, Taghavi S, Ajamian F. Investigating association of G103T polymorphism of Coagulation factor XIII and recurrent pregnancy loss. Iran J Reprod Med 2015; 4 (Suppl.): 53–54.
 Zahraei M, Sheikhha MH, Kalantar SM, Ghasemi N, Jahaninejad T, Rajabi SH, et al. The association of arylendosulfatase 1 (SULF1) gene polymorphism with recurrent miscarriage. J Assist Reprod Genet 2014; 31: 157–161.
 Pourmasumi S, Ghasemi N, Talebi AR, Mehrabani M, Sabeti P. The effect of vitamin E and selenium on sperm chromatin quality in couples with recurrent miscarriage. IJML 2018; 5: 1–10.
 Soleimanian S, Kalantar SM, Sheikhha MH, Zaimy MA, Rasti A, Fazli H. Association between Ychromosome AZFc region micro-deletions with recurrent miscarriage. Iran J Reprod Med 2013; 11: 431–434.
 Lotfabadi NN, Kouchesfahani HM, Sheikhha MH, Kalantar SM. In vitro transfection of anti-tumor miR- 101 induces BIM, a pro-apoptotic protein, expression in acute myeloid leukemia (AML). EXCLI J 2017; 16: 1257–1267.
 Lotfabadi NN, Kouchesfehani HM, Sheikhha MH, Kalantar SM. Development of a novel cationic liposome: Evaluation of liposome mediated transfection and anti-proliferative effects of miR- 101 in acute myeloid leukemia. J Drug Deliv Sci Technol 2018; 45: 196–202.
 Moghimi B, Gharibi S, Farashahi Yazd E, Taher Tahoori M, Kalantar SM. Role of miR-146a in immune system and autoimmunity. IJML 2016; 3: 150–158.
 Dorfeshan P, Novin MG, Salehi M, Farifteh F. Expression of miR-302 in human embryo derived from in-vitro matured oocyte. Int J Reprod Biomed 2019; 17: 405–412.
 Abdalla M, Deshmukh H, Atkin SL, Sathyapalan T. MiRNAs as a novel clinical biomarker and therapeutic targets in polycystic ovary syndrome (PCOS): A review. Life Sci 2020; 259: 118174.
 Da Broi MG, Meola J, Plaça JR, Peronni KC, Rocha CV, Silva WA, et al. Is the profile of transcripts altered in the eutopic endometrium of infertile women with endometriosis during the implantation window? Human Reprod 2019; 34: 2381–2390.
 Ahn DH, Rah HC, Choi YK, Jeon YJ, Min KT, Kwack K, et al. Association of the miR−146aC> G, miR−149T> C, miR−196a2T> C, and miR−499A> G polymorphisms with gastric cancer risk and survival in the korean population. Mol Carcinog 2013; 52: E39–E51.
 Hu Y, Liu CM, Qi L, He TZ, Shi-Guo L, Hao CJ, et al. Two common SNPs in pri-miR-125a alter the mature miRNA expression and associate with recurrent pregnancy loss in a Han-Chinese population. RNA Biol 2011; 8: 861–872.
 Scott GK, Goga A, Bhaumik D, Berger CE, Sullivan CS, Benz CC. Coordinate suppression of ERBB2 and ERBB3 by enforced expression of micro-RNA miR- 125a or miR-125b. J Biol Chem 2007; 282: 1479– 1486.
 Vo DT, Karanam NK, Ding L, Saha D, Yordy JS, Giri U, et al. miR-125a-5p functions as tumor suppressor microRNA and is a marker of locoregional recurrence and poor prognosis in head and neck cancer. Neoplasia 2019; 21: 849–862.
 Suzuki Y, Kim HW, Ashraf M, Haider KH. Diazoxide potentiates mesenchymal stem cell survival via NF- κB-dependent miR-146a expression by targeting Fas. Am J Physiol Heart Circ Physiol 2010; 299: H1077– H1082.
 Huang JB, Mei J, Jiang LY, Jiang ZL, Liu H, Zhang JW, et al. MiR-196a2 rs11614913 T> C polymorphism is associated with an increased risk of Tetralogy of Fallot in a Chinese population. Acta Cardiol Sin 2015; 31: 18–23.
 Fluhr H, Wenig H, Spratte J, Heidrich S, Ehrhardt J, Zygmunt M. Non-apoptotic Fas-induced regulation of cytokines in undifferentiated and decidualized human endometrial stromal cells depends on caspase-activity. Mol Hum Reprod 2010; 17: 127–134.
 Parveen F, Agrawal S. Recurrent miscarriage and micro-RNA among north Indian women. Reprod Sci 2015; 22: 410–415.
 Bortone F, Scandiffio L, Marcuzzo S, Bonanno S, Frangiamore R, Motta T, et al. MiR-146a in myasthenia gravis thymus bridges innate immunity with autoimmunity and is linked to therapeutic effects of corticosteroids. Front Immunol 2020; 11: 142–157.
 Kuang W, Zheng L, Xu X, Lin Y, Lin J, Wu J, et al. Dysregulation of the miR-146a-Smad4 axis impairs osteogenesis of bone mesenchymal stem cells under inflammation. Bone Res 2017; 5: 17037.
 Pu W, Shang Y, Shao Q, Yuan X. miR-146a promotes cell migration and invasion in melanoma by directly targeting SMAD4. Oncol Lett 2018; 15: 7111–7117.
 Fazli M, Ghorbian S. Association study of non-coding RNA miR-499 and miR196a2 gene polymorphisms with the risk of idiopathic recurrent pregnancy loss. Gene Cell and Tissue 2018; 5: 1–5.
 Lee JY, Kim JO, Park HS, Ryu CS, Kim JH, Kim YR, et al. Study of the association between microRNA (miR-25T> C, miR-32C> A, miR-125C> T, and miR- 222G> T) polymorphisms and the risk of recurrent pregnancy loss in korean women. Genes (Basel) 2020; 11: 354–366.
 Shaker M, Shalabi T, Gaber KR, Amr K. Association of miRNA-27a and leptin polymorphisms with recurrent pregnancy loss in Egyptian women. Meta Gene 2020; 24: 100617.
 Cho SH, An HJ, Kim KA, Ko JJ, Kim JH, Kim YR, et al. Single nucleotide polymorphisms at miR-146a/196a2 and their primary ovarian insufficiency-related target gene regulation in granulosa cells. PloS One 2017; 12: e0183479.
 Cho SH, Chung KW, Kim JO, Jang H, Yoo JK, Choi Y, et al. Association of miR-146aC> G, miR-149C> T, miR-196a2T> C, and miR-499A> G polymorphisms with risk of recurrent implantation failure in Korean women. Eur J Obstet Gynecol Reprod Biol 2016; 202: 14–19.
 Lim J, Kim JO, Park HS, Han IB, Kwack K, Kim NK, et al. Associations of miR-146aC> G, miR-149C> T, miR-196a2C> T and miR-499A> G polymorphisms with brain tumors. Oncol Rep 2018; 40: 1813–1823.
 Kim YR, Ryu CS, Kim JO, An HJ, Cho SH, Ahn EH, et al. Association study of AGO1 and AGO2 genes polymorphisms with recurrent pregnancy loss. Sci Rep 2019; 9: 15591.
 Turienzo A, Lledó B, Ortiz JA, Morales R, Sanz J, Llácer J, et al. Prevalence of candidate single nucleotide polymorphisms on p53, IL-11, IL-10, VEGF and APOE in patients with repeated implantation failure (RIF) and pregnancy loss (RPL). Hum Fertil 2018; 23: 117–122.
 Chai C, Wu H, Wang B, Eisenstat DD, Leng RP. MicroRNA-498 promotes proliferation and migration by targeting the tumor suppressor PTEN in breast cancer cells. Carcinogenesis 2018; 39: 1185–1196.
 Jairajpuri DS, Malalla ZH, Mahmood N, Khan F, Almawi WY. Differentially expressed circulating microRNAs associated with idiopathic recurrent pregnancy loss. Gene 2020; 768: 145334.
 Hussein MH, Toraih EA, Aly NM, Riad E, Fawzy MS. A passenger strand variant in miR-196a2 contributes to asthma severity in children and adolescents: A preliminary study. Biochem Cell Biol 2016; 94: 347– 357.
 Yu K, Ji Y, Wang H, Xuan Q, Li B, Xiao J, et al. Association of miR-196a2, miR-27a, and miR- 499 polymorphisms with isolated congenital heart disease in a Chinese population. Genet Mol Res 2016; 15: 1–14.