Juliana Carron , Caroline Torricelli , Janet Keller Silva , Lilian de Oliveira Coser , Carmen Silvia Passos Lima , Gustavo Jacob Lourenço
{"title":"MITF (rs7623610)和CREB1 (rs10932201)基因的内含子变异可能提高人类黑色素瘤细胞系的剪接效率","authors":"Juliana Carron , Caroline Torricelli , Janet Keller Silva , Lilian de Oliveira Coser , Carmen Silvia Passos Lima , Gustavo Jacob Lourenço","doi":"10.1016/j.mrfmmm.2021.111763","DOIUrl":null,"url":null,"abstract":"<div><p><span>We previously reported that intronic single nucleotide variations (SNVs) in </span><span><em>MITF</em></span> (c.938−325G>A, rs7623610) and <span><em>CREB1</em></span><span> (c.303+373G>A, rs10932201) genes were associated with risk, aggressiveness, and prognosis of cutaneous melanoma<span> (CM). In this study, we investigated the influence of the above SNVs in splicing patterns and efficiency. We constructed minigenes with wild type and variant alleles from </span></span><em>MITF</em> and <em>CREB1</em><span><span> to assess the effect of the SNVs on splicing. The minigenes were transfected in the human melanoma cell line (SK-MEL-28). RT-PCR and </span>DNA sequencing investigated the constructs’ splicing patterns. Minigenes constructs’ splicing efficiency and </span><em>HNRNPA1</em> and <em>SF1</em> splicing genes’ expression were investigated by qPCR. We found that <em>MITF</em> and <em>CREB1</em> SNVs did not alter the splicing pattern, but they influenced the splicing efficiency. <em>MITF</em>-A (<em>p=</em> 0.03) and <em>CREB1</em>-A (<em>p</em>= 0.005) variant minigenes yielded an increase of mRNA generated from the constructions. Additionally, lower mRNA levels of <em>HNRNPA1</em> and <em>SF1</em> were seen in the variant minigenes <em>MITF</em>-A (<em>p</em>= 0.04) and <em>CREB1</em>-A (<em>p</em>= 0.005). We described for the first time the potential importance of <em>MITF</em> rs7623610 and <em>CREB1</em> rs10932201 SNVs in splicing efficiency and its relationship with CM.</p></div>","PeriodicalId":49790,"journal":{"name":"Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis","volume":"823 ","pages":"Article 111763"},"PeriodicalIF":1.5000,"publicationDate":"2021-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Intronic variants of MITF (rs7623610) and CREB1 (rs10932201) genes may enhance splicing efficiency in human melanoma cell line\",\"authors\":\"Juliana Carron , Caroline Torricelli , Janet Keller Silva , Lilian de Oliveira Coser , Carmen Silvia Passos Lima , Gustavo Jacob Lourenço\",\"doi\":\"10.1016/j.mrfmmm.2021.111763\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p><span>We previously reported that intronic single nucleotide variations (SNVs) in </span><span><em>MITF</em></span> (c.938−325G>A, rs7623610) and <span><em>CREB1</em></span><span> (c.303+373G>A, rs10932201) genes were associated with risk, aggressiveness, and prognosis of cutaneous melanoma<span> (CM). In this study, we investigated the influence of the above SNVs in splicing patterns and efficiency. We constructed minigenes with wild type and variant alleles from </span></span><em>MITF</em> and <em>CREB1</em><span><span> to assess the effect of the SNVs on splicing. The minigenes were transfected in the human melanoma cell line (SK-MEL-28). RT-PCR and </span>DNA sequencing investigated the constructs’ splicing patterns. Minigenes constructs’ splicing efficiency and </span><em>HNRNPA1</em> and <em>SF1</em> splicing genes’ expression were investigated by qPCR. We found that <em>MITF</em> and <em>CREB1</em> SNVs did not alter the splicing pattern, but they influenced the splicing efficiency. <em>MITF</em>-A (<em>p=</em> 0.03) and <em>CREB1</em>-A (<em>p</em>= 0.005) variant minigenes yielded an increase of mRNA generated from the constructions. Additionally, lower mRNA levels of <em>HNRNPA1</em> and <em>SF1</em> were seen in the variant minigenes <em>MITF</em>-A (<em>p</em>= 0.04) and <em>CREB1</em>-A (<em>p</em>= 0.005). We described for the first time the potential importance of <em>MITF</em> rs7623610 and <em>CREB1</em> rs10932201 SNVs in splicing efficiency and its relationship with CM.</p></div>\",\"PeriodicalId\":49790,\"journal\":{\"name\":\"Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis\",\"volume\":\"823 \",\"pages\":\"Article 111763\"},\"PeriodicalIF\":1.5000,\"publicationDate\":\"2021-07-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0027510721000269\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"BIOTECHNOLOGY & APPLIED MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0027510721000269","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
Intronic variants of MITF (rs7623610) and CREB1 (rs10932201) genes may enhance splicing efficiency in human melanoma cell line
We previously reported that intronic single nucleotide variations (SNVs) in MITF (c.938−325G>A, rs7623610) and CREB1 (c.303+373G>A, rs10932201) genes were associated with risk, aggressiveness, and prognosis of cutaneous melanoma (CM). In this study, we investigated the influence of the above SNVs in splicing patterns and efficiency. We constructed minigenes with wild type and variant alleles from MITF and CREB1 to assess the effect of the SNVs on splicing. The minigenes were transfected in the human melanoma cell line (SK-MEL-28). RT-PCR and DNA sequencing investigated the constructs’ splicing patterns. Minigenes constructs’ splicing efficiency and HNRNPA1 and SF1 splicing genes’ expression were investigated by qPCR. We found that MITF and CREB1 SNVs did not alter the splicing pattern, but they influenced the splicing efficiency. MITF-A (p= 0.03) and CREB1-A (p= 0.005) variant minigenes yielded an increase of mRNA generated from the constructions. Additionally, lower mRNA levels of HNRNPA1 and SF1 were seen in the variant minigenes MITF-A (p= 0.04) and CREB1-A (p= 0.005). We described for the first time the potential importance of MITF rs7623610 and CREB1 rs10932201 SNVs in splicing efficiency and its relationship with CM.
期刊介绍：
Mutation Research (MR) provides a platform for publishing all aspects of DNA mutations and epimutations, from basic evolutionary aspects to translational applications in genetic and epigenetic diagnostics and therapy. Mutations are defined as all possible alterations in DNA sequence and sequence organization, from point mutations to genome structural variation, chromosomal aberrations and aneuploidy. Epimutations are defined as alterations in the epigenome, i.e., changes in DNA methylation, histone modification and small regulatory RNAs.
MR publishes articles in the following areas:
Of special interest are basic mechanisms through which DNA damage and mutations impact development and differentiation, stem cell biology and cell fate in general, including various forms of cell death and cellular senescence.
The study of genome instability in human molecular epidemiology and in relation to complex phenotypes, such as human disease, is considered a growing area of importance.
Mechanisms of (epi)mutation induction, for example, during DNA repair, replication or recombination; novel methods of (epi)mutation detection, with a focus on ultra-high-throughput sequencing.
Landscape of somatic mutations and epimutations in cancer and aging.
Role of de novo mutations in human disease and aging; mutations in population genomics.
Interactions between mutations and epimutations.
The role of epimutations in chromatin structure and function.
Mitochondrial DNA mutations and their consequences in terms of human disease and aging.
Novel ways to generate mutations and epimutations in cell lines and animal models.
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