title: MiR-196a binding-site SNP regulates RAP1A expression contributing to esophageal squamous cell carcinoma risk and metastasis.
abstract: Polymorphisms in 3' untranslated region (UTR) of cancer-related genes might affect regulation by microRNA (miRNA) and contribute to carcinogenesis. In this study, we screened several single nucleotide polymorphisms (SNPs) in 3'UTR of cancer-related genes and investigated their effects on the risk of esophageal squamous cell carcinoma (ESCC). First, we used SNaPshot assay to genotype seven 3'UTR SNPs in 537 ESCC cases and 608 normal controls in a Chinese Han population and found that SNP rs6573 in 3'UTR of RAS-related proteins (RAP1A) was significantly associated with ESCC risk [P = 0.02, odds ratio (OR) = 0.43; 95% confidence interval (CI): 0.21-0.91] and pathologic stage (P = 0.03, OR = 1.89; 95% CI: 1.06-3.36). A putative binding site for miRNA-196a (miR-196a) exists in the 3'UTR of RAP1A, and the genetic variant, rs6573 A→C, is present in this binding region. We confirmed that miR-196a regulated the expression of RAP1A by luciferase reporter assay and that the regulation was affected by the RAP1A genotype. SNP rs6573 A to C change interfere in the interaction of miR-196a binding to RAP1A 3'UTR, resulting in higher constitutive expression of RAP1A. Moreover, we observed that RAP1A was overexpressed in the majority of ESCC tissues and correlated with RAP1A genotype and lymph node metastasis. In vitro study indicated RAP1A might function as a promoter for esophageal cancer cell migration and invasion through matrix metalloproteinase 2. Our study highlights RAP1A and SNP rs6573 functioning as potential personal diagnostic and prognosis markers for ESCC.
authors.affiliation: Department of Medical Genetics, Third Military Medical University, Chongqing 400038, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Wang
authors.forename: Juan
authors.initials: J
authors.lastname: Li
authors.forename: Hong
authors.initials: H
authors.lastname: Guo
authors.forename: Xueqing
authors.initials: X
authors.lastname: Xu
authors.forename: Gang
authors.initials: G
authors.lastname: Xiong
authors.forename: Xingying
authors.initials: X
authors.lastname: Guan
authors.forename: Botao
authors.initials: B
authors.lastname: Liu
authors.forename: Junxia
authors.initials: J
authors.lastname: Li
authors.forename: Xuedan
authors.initials: X
authors.lastname: Chen
authors.forename: Kang
authors.initials: K
authors.lastname: Yang
authors.forename: Yun
authors.initials: Y
authors.lastname: Bai
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics*; D001665:Binding Sites; D015153:Blotting, Western; D002294:Carcinoma, Squamous Cell / Q000209:etiology* / Q000378:metabolism / Q000556:secondary; D016022:Case-Control Studies; D002453:Cell Cycle; D002465:Cell Movement; D049109:Cell Proliferation; D004938:Esophageal Neoplasms / Q000209:etiology* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D008207:Lymphatic Metastasis; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D009367:Neoplasm Staging; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D011379:Prognosis; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020827:rap1 GTP-Binding Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D020413:3' Untranslated Regions; C542543:MIRN196 microRNA, human; D035683:MicroRNAs; C430803:RAP1A protein, human; D012333:RNA, Messenger; D008156:Luciferases; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2; D020827:rap1 GTP-Binding Proteins