lncRNA人类结肠癌相关转移因子1 SNPs与胃癌遗传易感性关联研究

发布时间：2018-03-20 14:26

本文选题：长链非编码　切入点：RNA　出处：《郑州大学》2017年硕士论文　论文类型：学位论文

【摘要】：胃癌在全世界范围是严重危害人类健康的常见的恶性肿瘤之一,发病率占全部恶性肿瘤的第4位,占消化道恶性肿瘤的首位。据统计,70%的胃癌病例发生在发展中国家,其中中国人群病例占据将近45%,五年生存率仍不容乐观,病人生活质量较差及家庭社会经济负担较重,胃癌的发病机制的深入探索及研究仍是亟待解决的。随着胃癌病因学以及胃癌发生、发展机制的逐步深入研究,我们发现基因与环境因素均起着至关重要的作用。随着研究的不断深入,长链非编码RNA(long noncoding RNAs,lnc RNAs)在癌症的发生发展机制中有着重要的调控作用,越来越多的lnc RNA与胃癌存在密切的关联。人类结肠癌相关转移因子-1(colon cancer associated transcript-1,CCAT1)在胃癌组织中高表达,且与肿瘤的侵袭、转移及预后密切相关。lnc RNA遗传多态性在胃癌易感性机制研究提供重要突破。目的研究CCAT1单核苷酸多态性(SNPs)(rs77628730,rs6989575,rs7816475以及rs6470502)与胃癌易感性的关系,并进一步探索lnc RNA CCAT1 SNPs与环境因素之间是否存在交互作用,对存在关联的位点进行初步功能实验验证,为发现新的胃癌遗传易感位点提供理论支持,为胃癌的早发现以及早期诊断提供可靠的分子标志物。方法查阅国内外文献数据库以及lnc RNA数据库,运用生物信息学网站及软件筛选CCAT1上的标签SNPs。该研究采用病例-对照方法,选取来源于医院的460例经病理学诊断确诊的胃癌患者作为病例组,同时将性别及年龄按照频数匹配选取来源于社区的正常对照460例。对rs77628730以及rs6470502两SNPs位点运用聚合酶链式反应-限制性片段长度分析法(PCR-RFLP)对进行基因分型,rs6989575以及rs7816475采用创造性酶切位点分析法(CRS-PCR)。通过SPSS21.0软件对病例组与对照组的一般特征、四个位点不同基因型以及等位基因的分布频率之间的差异采用t检验以及χ2检验;通过Hardy-Weinberg在线分析软件对对照组基因型频率进行Hardy-Weinberg平衡分析;单体型分析通过SHEsis在线软件进行;通过MDR2.0软件对基因-环境交互作用进行分析。对正常人群中有意义位点CCAT1 rs6470502不同基因型的血清中CCAT1mRNA表达量的差异通过q RT-PCR检测,进一步验证CCAT1 rs6470502遗传变异是否影响CCAT1 mRNA的表达量。进一步探讨其在胃癌发生发展过程中的调控机制。结果CCAT1 rs6470502位点SNP与胃癌密切相关,与CC纯合基因型相比,TT纯合基因型(OR调整:1.68;95%CI:1.01-2.79)在调整吸烟,饮酒及家族史这些混杂因素后能显著增加胃癌的发病风险;相较于C等位基因,T等位基因与胃癌的风险增加显著相关(OR:1.94;95%CI:1.53-2.46)。相较于A等位基因而言,rs7816475位点G等位基因与胃癌的风险降低显著相关(OR:0.78;95%CI:0.62-0.99)。按年龄、性别、吸烟史、饮酒史及胃癌家族史进行分层分析,多因素logistic回归分析结果显示,rs6470502位点在大于50岁年龄段人群中(OR=1.69,95%CI=1.01-2.98)以及吸烟者(OR=1.95,95%CI=1.41-2.89)中患胃癌的发病风险有不同程度的增加;rs7816475位点在胃癌家族史者(OR=1.74,95%CI=1.30-2.87)中患胃癌风险均呈现不同程度的增加。q RT-PCR结果显示,CCAT1 rs6470502位点TT(2.08±0.55)、TC(1.70±0.38)和TC+TT(1.95±0.40)基因型中CCAT1 mRNA的表达量均高于rs6470502CC(1.16±0.40)基因型,差异均具有统计学意义(P0.05)。基因-环境因素交互作用分析结果显示携带rs77628730 A等位基因,rs6989575 T等位基因以及rs6470502 T等位基因且具有吸烟史人群患胃癌的风险显著增加(OR:3.83;95%CI:2.96-4.96)。结论1.CCAT1 rs6470502突变等位基因T以及TT基因型可显著增加胃癌发病风险,rs7816475突变等位基因G可降低胃癌的发病风险。2.CCAT1 rs6470502 CT+TT基因型与年龄50岁相关以及吸烟者存在相关,rs7816475 AG+GG基因型与肿瘤家族史之间存在关联。3.CCAT1基因单体型A_(rs77628730)C_(rs6989575)G_(rs7816475)T_(rs6470502)、T_(rs77628730)C_(rs6989575)G_(rs7816475)C_(rs6470502)、T_(rs77628730)T_(rs6989575)A_(rs7816475)C_(rs6470502)可以降低胃癌发病风险,单体型A_(rs77628730)T_(rs6989575)G_(rs7816475)T_(rs6470502)、T_(rs77628730)C_(rs6989575)A_(rs7816475)T_(rs6470502)可增加胃癌发病风险。4.CCAT_(1 rs6470502)基因突变可能调节CCAT1 mRNA表达量。5.吸烟史、rs77628730 A等位基因、rs6989575 T等位基因以及rs6470502 T等位基因在胃癌发生中存在交互作用。
[Abstract]:Gastric cancer is one of the serious harm to human health of the common malignant tumors in the world, the incidence rate of malignant tumor accounted for fourth, accounting for malignant tumors of the digestive tract in the first place. According to statistics, 70% gastric cancer cases occur in developing countries, which China cases occupy nearly 45% people, the five year survival rate is still not optimistic, patient life the poor quality of family and social economic burden is heavy, still further explore and study the pathogenesis of gastric cancer to be solved. With gastric cancer etiology and pathogenesis of gastric cancer, through in-depth study on development mechanism, which plays a vital role in our discovery of genetic and environmental factors. With the deepening of the study of long chain non encoding (RNA long noncoding RNAs, LNC RNAs) plays an important role in the regulation of the mechanism of the occurrence and development of cancer, gastric cancer and LNC RNA more and more closely related to human colon cancer. Related factor -1 (colon cancer associated transfer transcript-1, CCAT1) high expression in gastric cancer tissue, and tumor invasion, metastasis and prognosis of.Lnc is closely related to the genetic polymorphism of RNA provides an important breakthrough in the study of susceptibility to gastric cancer. Objective to study the mechanism of CCAT1 single nucleotide polymorphism (SNPs) (rs77628730, rs6989575, rs7816475 and rs6470502) relationship susceptibility to gastric cancer, and to further explore the interaction between LNC RNA CCAT1 SNPs and environmental factors, the preliminary function of experimental verification of the associated site, to discover new genetic susceptibility loci for gastric cancer and provide theoretical support to provide reliable molecular markers for early detection and early diagnosis of gastric cancer. Methods at home and abroad the literature database and LNC RNA database, using bioinformatics software and web site screening by case - control the label on the CCAT1 SNPs. of the study Methods selected from hospital in 460 cases confirmed by pathological diagnosis of gastric cancer patients as case group, while gender and age in normal control, frequency selection of sources in the community. In 460 cases of rs77628730 and rs6470502 two SNPs loci by analysis of polymerase chain reaction restriction fragment length (PCR-RFLP) of the gene type rs6989575 and rs7816475 by enzyme digestion sites creative analysis method (CRS-PCR). Based on the SPSS21.0 software, the case group and the control group the general characteristics, the difference between the distribution frequency of four loci and alleles by t test and 2 test; analysis software of genotypic frequency in control group was Hardy-Weinberg balance analysis online through Hardy-Weinberg; haplotype analysis was performed by SHEsis online software; gene environment interaction was analyzed by MDR2.0 software. The difference of expression level by Q RT-PCR to detect serum CCAT1mRNA significant loci CCAT1 rs6470502 genotypes in normal population, further verify the genetic variation of rs6470502 CCAT1 influence expression of CCAT1 mRNA. To further explore the process in the development of gastric cancer in the control mechanism. The results are closely related to the CCAT1 site of rs6470502 SNP and gastric cancer, compared with CC homozygous genotype, TT homozygous genotype (OR: 1.68; 95%CI:1.01-2.79) after adjusting for smoking, drinking and family history of these confounding factors could significantly increase the risk of gastric cancer; compared to the C allele, T allele increased the risk of gastric cancer and significantly correlated (OR:1.94; 95%CI:1.53-2.46) compared. In the A allele, rs7816475 risk allele G and gastric cancer decreased significantly correlated (OR:0.78; 95%CI:0.62-0.99). According to age, gender, smoking history, drinking history and familial gastric cancer The history of stratified analysis, multivariate logistic regression analysis showed that the rs6470502 locus in more than 50 years of age (OR=1.69,95%CI=1.01-2.98) and smokers (OR=1.95,95%CI=1.41-2.89) risk in gastric cancer have different degrees of increase; rs7816475 in the family history of gastric cancer (OR=1.74,95%CI=1.30-2.87) in gastric cancer risk were increased in different degrees.Q RT-PCR results showed that the CCAT1 rs6470502 site of TT (2.08 + 0.55), TC (1.70 + 0.38) and TC+TT (1.95 + 0.40) gene expression amount of CCAT1 in type mRNA was higher than that of rs6470502CC (1.16 + 0.40) genotypes, the differences were statistically significant (P0.05). The gene environment interaction analysis display rs77628730 carrying A allele rs6989575, T allele and rs6470502 T allele and has a history of smoking people suffering from gastric cancer risk increased significantly (OR:3.83; 95%CI:2.96- 4.96). Conclusion 1.CCAT1 rs6470502 mutant T allele and TT genotype significantly increased the risk of gastric cancer, rs7816475 mutant allele G can reduce the risk of.2.CCAT1 rs6470502 CT+TT genotype and age of gastric cancer at the age of 50 and the related association of smokers, the haplotypes of.3.CCAT1 gene between A_ rs7816475 genotype and AG+GG family history the tumor (rs77628730) C_ (rs6989575) G_ (rs7816475) T_ (rs6470502), T_ (rs77628730) C_ (rs6989575) G_ (rs7816475) C_ (rs6470502), T_ (rs77628730) T_ (rs6989575) A_ (rs7816475) C_ (rs6470502) can reduce the risk of gastric cancer, haplotype A_ (rs77628730) (T_ rs6989575) G_ (rs7816475) T_ (rs6470502), T_ (rs77628730) C_ (rs6989575) A_ (rs7816475) T_ (rs6470502) can increase the risk of gastric cancer (.4.CCAT_ rs6470502 1) gene mutation may regulate the expression of mRNA CCAT1.5. rs77628730 A smoking history. The allele, rs6989575 T allele and rs6470502 T allele have interaction in the occurrence of gastric cancer.

【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.2

【相似文献】