青岛地区汉族人群中lncRNA H19的多态性与胃癌和EBV相关胃癌易感性的关系
|
摘要
目的:探讨青岛地区汉族人群lncRNA H19单核苷酸多态性(SNP)与胃癌和EBV相关胃癌(EBVaGC)易感性的关系。方法:收集青岛地区汉族人群2015年1月至2018年10月青岛大学附属医院经病理科确诊为胃癌的新鲜组织或陈旧的石蜡包埋胃癌组织病理标本共225例,为胃癌组;依据原位杂交法对EBV编码的小分子非多聚腺苷酸(EBER1)转录检测结果再将胃癌组分为2亚组:EBVaGC组70例,EBVnGC组155例;同时选择青岛大学附属医院门诊健康体检者200例为对照组。提取EBVaGC、EBVnGC组织及健康人群外周血标本的DNA,根据HaploView软件常规设置原则(MAF>0.05;r2>0.8)筛选出rs217727、rs2735971、rs2839698和rs3741216四个H19的TagSNPs。利用Taq-Man MGB等位基因分型试剂盒对各SNP位点基因进行基因分型,并进行基因多态性检测。结果:所取标本的H19 SNPs均符合Hardy-Weinberg平衡。与对照组比较,胃癌组H19 rs217727位点TT基因型的发病风险显著增加(χ~2=9.073, P=0.003, OR=1.999, 95%CI=1.271～3.143),等位基因T的分布也明显增高(χ~2=13.475, P=0.001, OR=1.661, 95%CI=1.266～2.180);H19 rs2839698位点TC、CC基因型人群可显著增加胃癌的发病风险(χ~2=9.407,P=0.002;χ~2=6.517, P=0.011),携带C等位基因人群罹患胃癌的风险明显增加(χ~2=6.163, P=0.013, OR=1.417, 95%CI=1.076～1.867;χ~2=9.542, P=0.02, OR=2.070, 95%CI=1.298～3.302)。但胃癌组H19 rs2735971和rs3741216位点基因多态性与对照组比较差异不明显(均P>0.05);EBVaGC和EBVnGC组中H19的4个位点基因多态性分布差异均无统计学意义(均P>0.05)。结论:H19rs217727、rs2839698基因多态性可能与胃癌发病风险有关,携带TT基因型C等位基因和人群胃癌的发病风险明显升高;H19SNP的多态性与EBVaGC的发病风险无明显相关。
Objective: To evaluate the potential associations between the single nucleotide polymorphisms(SNPs) of lncRNA H19 genes and the susceptibility to gastric cancer(GC), especially to EBV-associated gastric cancer(EBVaGC) in Han population in Qingdao. Methods: 225 cases of pathologically confirmed fresh gastric cancer tissues or paraffin embedded gastric cancer tissues during January 2015 and October 2018 were collected from Affiliated Hospital of Qingdao University, as GC group; in the meanwhile, 200 healthy people underwent physical examination at Outpatient were collected as control. The 225 cases of cancer tissues were assigned into two groups according to the transcription result of EBV-encoded small molecule non-polyadenylation(EBER 1) by In situ hybridization:EBVaGC group(70 cases) and EBVnGC group(155 cases). DNA was extracted from the tissues of two groups and peripheral blood of healthy participants. According to the general setting of HaploView software(MAF>0.05, r2>0.8), four TagSNPs(rs217727, rs2735971,rs2839698 and rs3741216) of H19 were screened. Genotyping of each SNP locus was carried out by using Taq-Man MGB allele typing kit, and the gene polymorphisms were examined. Results: H19 SNPs of collected tissue samples were in accordance with the HardyWeinberg equilibrium. Compared with control group, patients carrying TT genotype of H19 rs217727 loci had significantly increased susceptibility to gastric cancer(χ~2=9.073, P=0.003, OR=1.999, 95% CI=1.271-3.143), so did the T allele(χ~2=13.475, P=0.001, OR=1.661, 95% CI=1.266-2.180). In contrast, patients carrying TC and CC genotype of rs2839698 loci had significantly increased susceptibility to gastric cancer(χ~2=9.407, P=0.002; χ~2=6.517, P=0.011), and patients carrying C allele had obviously increased susceptibility to GC(χ~2=6.163, P=0.013, OR=1.417, 95%CI=1.076-1.867; χ~2=9.542, P=0.02, OR=2.070, 95%CI=1.298-3.302). As for the rs2735971 and rs3741216, there was no significant difference between the GC group and the control group(all P>0.05). The distribution of 4 H19 SNPs showed no statistical difference in both EBVaGC group and EBVnGC group(all P>0.05). Conclusion: There was an association between H19 gene polymorphism(rs217727 and rs2839698) and gastric carcinoma; patients carrying the TT genotype of rs217727 and C allele of rs2839698 may increase the risk of gastric carcinoma. No significant association was observed between H19 SNP polymorphism and risk of EBVaGC.
Objective: To evaluate the potential associations between the single nucleotide polymorphisms(SNPs) of lncRNA H19 genes and the susceptibility to gastric cancer(GC), especially to EBV-associated gastric cancer(EBVaGC) in Han population in Qingdao. Methods: 225 cases of pathologically confirmed fresh gastric cancer tissues or paraffin embedded gastric cancer tissues during January 2015 and October 2018 were collected from Affiliated Hospital of Qingdao University, as GC group; in the meanwhile, 200 healthy people underwent physical examination at Outpatient were collected as control. The 225 cases of cancer tissues were assigned into two groups according to the transcription result of EBV-encoded small molecule non-polyadenylation(EBER 1) by In situ hybridization:EBVaGC group(70 cases) and EBVnGC group(155 cases). DNA was extracted from the tissues of two groups and peripheral blood of healthy participants. According to the general setting of HaploView software(MAF>0.05, r2>0.8), four TagSNPs(rs217727, rs2735971,rs2839698 and rs3741216) of H19 were screened. Genotyping of each SNP locus was carried out by using Taq-Man MGB allele typing kit, and the gene polymorphisms were examined. Results: H19 SNPs of collected tissue samples were in accordance with the HardyWeinberg equilibrium. Compared with control group, patients carrying TT genotype of H19 rs217727 loci had significantly increased susceptibility to gastric cancer(χ~2=9.073, P=0.003, OR=1.999, 95% CI=1.271-3.143), so did the T allele(χ~2=13.475, P=0.001, OR=1.661, 95% CI=1.266-2.180). In contrast, patients carrying TC and CC genotype of rs2839698 loci had significantly increased susceptibility to gastric cancer(χ~2=9.407, P=0.002; χ~2=6.517, P=0.011), and patients carrying C allele had obviously increased susceptibility to GC(χ~2=6.163, P=0.013, OR=1.417, 95%CI=1.076-1.867; χ~2=9.542, P=0.02, OR=2.070, 95%CI=1.298-3.302). As for the rs2735971 and rs3741216, there was no significant difference between the GC group and the control group(all P>0.05). The distribution of 4 H19 SNPs showed no statistical difference in both EBVaGC group and EBVnGC group(all P>0.05). Conclusion: There was an association between H19 gene polymorphism(rs217727 and rs2839698) and gastric carcinoma; patients carrying the TT genotype of rs217727 and C allele of rs2839698 may increase the risk of gastric carcinoma. No significant association was observed between H19 SNP polymorphism and risk of EBVaGC.
引文
[1]SIEGEL R L,MILLER K D,JEMAL A.Cancer statistics,2016[J].CA Cancer J Clin,2016,66(1):7-30.DOI:10.3322/caac.21332.
[2]ESTELLER M.Non-coding RNAs in human disease[J].Nat Rev Genet,2011,12(12):861-874.DOI:10.1038/nrg3074.
[3]QURESHI I A,MATTICK J S,MEHLER M F.Long non-coding RNAs in nervous system function and disease[J/OL].Brain Res,2010,1338:20-35[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2883659/.DOI:10.1016/j.brainres.2010.03.110.
[4]WAPINSKI O,CHANG H Y.Long non-coding RNAs and human disease[J].Trends Cell Biol,2011,21(6):354-361.DOI:10.1016/j.tcb.2011.04.001.
[5]ZHANG K J,LUO Z L,ZHANG Y,et al.Circulating lncRNA H19in plasma as a novel biomarker for breast cancer[J].Cancer Biomark,2016,17(2):187-194.DOI:10.3233/CBM-160630.[PubMed]
[6]HUANG C,CAO L H,QIU L M,et al.Upregulation of H19 promotes invasion and induces epithelial-to-mesenchymal transition in esophageal cancer[J/OL].Oncol Lett,2015,10(1):291-296[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4487103/.DOI:10.3892/ol.2015.3165.
[7]HAN D,GAO X,WANG M,et al.Long noncoding RNA H19 indicates a poor prognosis of colorectal cancer and promotes tumor growth by recruiting and binding to eIF4A3[J/OL].Oncotarget,2016,7(16):22159-22173[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5008352/.DOI:10.18632/oncotarget.8063.
[8]张俊国,林志丰,邓煜盛,等.肝癌遗传关联研究中LncRNA H19遗传变异的生物信息学分析[J].广东药学院学报,2015,31(6):816-819.
[9]Comprehensive molecular characterization of gastric adenocarcinoma[J/OL].Nature,2014,513(7517):202-209[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4170219/.DOI:10.1038/nature13480.
[10]GULLEY M L.Genomic assays for Epstein-Barr virus-positive gastric adenocarcinoma[J/OL].Exp Mol Med,2015,47:e134[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4314585/.DOI:10.1038/emm.2014.93.
[11]NISHIKAWA J,YOSHIYAMA H,IIZASA H,et al.Epstein-barr virus in gastric carcinoma[J].Cancers,2014,6(4):2259-2274.DOI:10.3390/cancers6042259.
[12]DU M L,WANG W Z,JIN H,et al.The association analysis of lnc RNA HOTAIR genetic variants and gastric cancer risk in a Chinese population[J].Oncotarget,2015,6(31):31255-31262..DOI:10.18632/oncotarget.5158.
[13]段然,颜成睿,王磊,等.lncRNA POU3F3通过调节MGMT的表达影响高级别脑胶质瘤细胞对替莫唑胺耐药[J].中国肿瘤生物治疗杂志,2019,26(3):328-332.10.3872/j.issn.1007-385X.2019.03.013.
[14]HAN D,GAO X,WANG M,et al.Long noncoding RNA H19 indicates a poor prognosis of colorectal cancer and promotes tumor growth by recruiting and binding to eIF4A3[J/OL].Oncotarget,2016,7(16):22159-22173[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5008352/.DOI:10.18632/oncotarget.8063
[15]MATOUK I J,DEGROOT N,MEZAN S,et al.The H19 non-coding RNA is essential for human tumor growth[J/OL].PLoS One,2007,2(9):e845[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1959184/.DOI:10.1371/journal.pone.0000845.
[16]NIINUMA T,SUZUKI H,NOJIMA M,et al.Upregulation of miR-196a and HOTAIR drive malignant character in gastrointestinal stromal tumors[J].Cancer Res,2012,72(5):1126-1136.DOI:10.1158/0008-5472.CAN-11-1803.
[17]LI H,YU B Q,LI J F,et al.Overexpression of lncRNA H19 enhances carcinogenesis and metastasis of gastric cancer[J/OL].Oncotarget,2014,5(8):2318-2329[20190-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4039165/.DOI:10.18632/oncotarget.1913.
[18]LI S W,HUA Y B,JIN J,et al.Association of genetic variants in lncRNA H19 with risk of colorectal cancer in a Chinese population[J/OL].Oncotarget,2016,7(18):25470-25477[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5041918/.DOI:10.18632/oncotarget.8330.
[19]LIN Y X,FU F M,CHEN Y Z,et al.Genetic variants in long noncoding RNA H19 contribute to the risk of breast cancer in a southeast China Han population[J/OL].Onco Targets Ther,2017,10:4369-4378[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5593399/.DOI:10.2147/OTT.S127962.
[20]YANG M L,HUANG Z,WANG Q,et al.The association of polymorphisms in lncRNA-H19 with hepatocellular cancer risk and prognosis[J/OL].Biosci Rep,2018,38(5):BSR20171652[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6123070/.DOI:10.1042/BSR20171652.
[21]VERHAEGH G W,VERKLEIJ L,VERMEULEN S H,et al.Polymorphisms in the H19 gene and the risk of bladder cancer[J].Eur Urol,2008,54(5):1118-1126.DOI:10.1016/j.eururo.2008.01.060.
[22]YANG C A,SCHEIBENBOGEN C,BAUER S,et al.A frequent Toll-like receptor 1 gene polymorphism affects NK-and T-cell IFN-γproduction and is associated with Helicobacter pylori-induced gastric disease[J].Helicobacter,2013,18(1):13-21.DOI:10.1111/hel.12001.
[23]Nishikawa J,Yoshiyama H,Iizasa H,et al.Epstein-barrvirus in gastric carcinoma[J].Cancers(Basel),2014,6(4):2259-2274.
[24]GULLEY M L.Genomic assays for Epstein-Barr virus-positive gastric adenocarcinoma[J/OL].Exp Mol Med,2015,47:e134[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4314585/.DOI:10.1038/emm.2014.93.
[2]ESTELLER M.Non-coding RNAs in human disease[J].Nat Rev Genet,2011,12(12):861-874.DOI:10.1038/nrg3074.
[3]QURESHI I A,MATTICK J S,MEHLER M F.Long non-coding RNAs in nervous system function and disease[J/OL].Brain Res,2010,1338:20-35[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2883659/.DOI:10.1016/j.brainres.2010.03.110.
[4]WAPINSKI O,CHANG H Y.Long non-coding RNAs and human disease[J].Trends Cell Biol,2011,21(6):354-361.DOI:10.1016/j.tcb.2011.04.001.
[5]ZHANG K J,LUO Z L,ZHANG Y,et al.Circulating lncRNA H19in plasma as a novel biomarker for breast cancer[J].Cancer Biomark,2016,17(2):187-194.DOI:10.3233/CBM-160630.[PubMed]
[6]HUANG C,CAO L H,QIU L M,et al.Upregulation of H19 promotes invasion and induces epithelial-to-mesenchymal transition in esophageal cancer[J/OL].Oncol Lett,2015,10(1):291-296[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4487103/.DOI:10.3892/ol.2015.3165.
[7]HAN D,GAO X,WANG M,et al.Long noncoding RNA H19 indicates a poor prognosis of colorectal cancer and promotes tumor growth by recruiting and binding to eIF4A3[J/OL].Oncotarget,2016,7(16):22159-22173[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5008352/.DOI:10.18632/oncotarget.8063.
[8]张俊国,林志丰,邓煜盛,等.肝癌遗传关联研究中LncRNA H19遗传变异的生物信息学分析[J].广东药学院学报,2015,31(6):816-819.
[9]Comprehensive molecular characterization of gastric adenocarcinoma[J/OL].Nature,2014,513(7517):202-209[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4170219/.DOI:10.1038/nature13480.
[10]GULLEY M L.Genomic assays for Epstein-Barr virus-positive gastric adenocarcinoma[J/OL].Exp Mol Med,2015,47:e134[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4314585/.DOI:10.1038/emm.2014.93.
[11]NISHIKAWA J,YOSHIYAMA H,IIZASA H,et al.Epstein-barr virus in gastric carcinoma[J].Cancers,2014,6(4):2259-2274.DOI:10.3390/cancers6042259.
[12]DU M L,WANG W Z,JIN H,et al.The association analysis of lnc RNA HOTAIR genetic variants and gastric cancer risk in a Chinese population[J].Oncotarget,2015,6(31):31255-31262..DOI:10.18632/oncotarget.5158.
[13]段然,颜成睿,王磊,等.lncRNA POU3F3通过调节MGMT的表达影响高级别脑胶质瘤细胞对替莫唑胺耐药[J].中国肿瘤生物治疗杂志,2019,26(3):328-332.10.3872/j.issn.1007-385X.2019.03.013.
[14]HAN D,GAO X,WANG M,et al.Long noncoding RNA H19 indicates a poor prognosis of colorectal cancer and promotes tumor growth by recruiting and binding to eIF4A3[J/OL].Oncotarget,2016,7(16):22159-22173[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5008352/.DOI:10.18632/oncotarget.8063
[15]MATOUK I J,DEGROOT N,MEZAN S,et al.The H19 non-coding RNA is essential for human tumor growth[J/OL].PLoS One,2007,2(9):e845[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1959184/.DOI:10.1371/journal.pone.0000845.
[16]NIINUMA T,SUZUKI H,NOJIMA M,et al.Upregulation of miR-196a and HOTAIR drive malignant character in gastrointestinal stromal tumors[J].Cancer Res,2012,72(5):1126-1136.DOI:10.1158/0008-5472.CAN-11-1803.
[17]LI H,YU B Q,LI J F,et al.Overexpression of lncRNA H19 enhances carcinogenesis and metastasis of gastric cancer[J/OL].Oncotarget,2014,5(8):2318-2329[20190-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4039165/.DOI:10.18632/oncotarget.1913.
[18]LI S W,HUA Y B,JIN J,et al.Association of genetic variants in lncRNA H19 with risk of colorectal cancer in a Chinese population[J/OL].Oncotarget,2016,7(18):25470-25477[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5041918/.DOI:10.18632/oncotarget.8330.
[19]LIN Y X,FU F M,CHEN Y Z,et al.Genetic variants in long noncoding RNA H19 contribute to the risk of breast cancer in a southeast China Han population[J/OL].Onco Targets Ther,2017,10:4369-4378[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5593399/.DOI:10.2147/OTT.S127962.
[20]YANG M L,HUANG Z,WANG Q,et al.The association of polymorphisms in lncRNA-H19 with hepatocellular cancer risk and prognosis[J/OL].Biosci Rep,2018,38(5):BSR20171652[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6123070/.DOI:10.1042/BSR20171652.
[21]VERHAEGH G W,VERKLEIJ L,VERMEULEN S H,et al.Polymorphisms in the H19 gene and the risk of bladder cancer[J].Eur Urol,2008,54(5):1118-1126.DOI:10.1016/j.eururo.2008.01.060.
[22]YANG C A,SCHEIBENBOGEN C,BAUER S,et al.A frequent Toll-like receptor 1 gene polymorphism affects NK-and T-cell IFN-γproduction and is associated with Helicobacter pylori-induced gastric disease[J].Helicobacter,2013,18(1):13-21.DOI:10.1111/hel.12001.
[23]Nishikawa J,Yoshiyama H,Iizasa H,et al.Epstein-barrvirus in gastric carcinoma[J].Cancers(Basel),2014,6(4):2259-2274.
[24]GULLEY M L.Genomic assays for Epstein-Barr virus-positive gastric adenocarcinoma[J/OL].Exp Mol Med,2015,47:e134[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4314585/.DOI:10.1038/emm.2014.93.