儿茶酚-O-甲基转移酶基因功能区单核苷酸多态性及甲基化水平与卵巢早衰的相关性
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摘要
目的探讨儿茶酚-O-甲基转移酶基因(COMT)4个功能区单核苷酸多态性(SNP)、甲基化水平及COMT基因mRNA表达与卵巢早衰(POF)的相关性。方法严格按照诊断标准选取304例POF患者为POF组,另317例健康者为对照组。采取静脉血,提取基因组DNA及RNA,采用SNaPshot技术检测4个SNPs的基因型,MethylTarget目标区域甲基化测序方法检测甲基化水平,RT-PCR检测COMT mRNA表达水平。结果 POF组rs4680位点AA基因型(OR=1.605, 95%CI=1.111～2.319,P=0.011)及A等位基因(OR=1.277,95%CI=1.022～1.596,P=0.032)频率显著高于对照组。POF组CpG_8(t=-2.234,P=0.027)和CpG_10(t=-2.033,P=0.043)甲基化水平显著高于对照组。POF组COMT基因mRNA表达低于对照组(t=2.071,P=0.041)。结论 COMT基因rs4680位点A等位基因及CpG_8和CpG_10甲基化水平的升高可能下调了COMT基因的表达,从而影响POF的发生发展。
Objective To evaluate the association of four single nucleotide polymorphisms(SNPs) in the functional regions, methylation and mRNA expression level of the catechol-Omethyl transferase(COMT) gene with the risk of premature ovarian failure(POF). Methods According to the POF diagnostic criteria, the participants recruited in this study included 304 patients with POF and 317 healthy women as controls. DNA and RNA were extracted from the peripheral blood. Genotyping of the 4 SNPs was conducted using the SNaPshot method. Methylation and mRNA level was tested by MethylTarget target region methylation sequencing and RT-PCR, respectively. Results The outcomes showed that the COMT gene rs4680 AA genotype(OR=1.605, 95% CI=1.111-2.319, P=0.011) and A allele(OR=1.277, 95% CI=1.022-1.596, P=0.032) frequencies, methylation level of CpG_8(t=-2.234, P=0.027) and CpG_10(t=-2.033, P=0.043) were significantly higher in the POF patients than in the healthy controls. The COMT gene mRNA level was significantly lower in the POF patients than the healthy controls(t=2.071,P=0.041).Conclusion Our findings indicate that the COMTgene rs4680 A allele,and methylation level of CpG_8 and CpG_10 may be associated with the risk and development of POF by downregulating the COMTgene expression level.
Objective To evaluate the association of four single nucleotide polymorphisms(SNPs) in the functional regions, methylation and mRNA expression level of the catechol-Omethyl transferase(COMT) gene with the risk of premature ovarian failure(POF). Methods According to the POF diagnostic criteria, the participants recruited in this study included 304 patients with POF and 317 healthy women as controls. DNA and RNA were extracted from the peripheral blood. Genotyping of the 4 SNPs was conducted using the SNaPshot method. Methylation and mRNA level was tested by MethylTarget target region methylation sequencing and RT-PCR, respectively. Results The outcomes showed that the COMT gene rs4680 AA genotype(OR=1.605, 95% CI=1.111-2.319, P=0.011) and A allele(OR=1.277, 95% CI=1.022-1.596, P=0.032) frequencies, methylation level of CpG_8(t=-2.234, P=0.027) and CpG_10(t=-2.033, P=0.043) were significantly higher in the POF patients than in the healthy controls. The COMT gene mRNA level was significantly lower in the POF patients than the healthy controls(t=2.071,P=0.041).Conclusion Our findings indicate that the COMTgene rs4680 A allele,and methylation level of CpG_8 and CpG_10 may be associated with the risk and development of POF by downregulating the COMTgene expression level.
引文
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[9] MOUFARRIJ S,DANDAPANI M,ARTHOFER E,et al.Epigenetic therapy for ovarian cancer:Promise and progress[J].Clin Epigenetics,2019,11(1):7.
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[11] HIRATA H,HINODA Y,OKAYAMA N,et al.COMT polymor-phisms affecting protein expression are risk factors for endometrial cancer[J].Mol Carcinog,2008,47 (10):768-774.
[12] DOHERTY JA,WEISS NS,FREEMAN RJ,et al.Genetic factors in catechol estrogen metabolism in relation to the risk of endometrial cancer[J].Cancer Epidemiol Biomarkers Prev,2005,14(2):357-366.
[13] LIU JX,LUO RC,LI R,et al.Lack of associations of the COMT Val158Met polymorphism with risk of endometrial and ovarian cancer:A pooled analysis of case-control studies[J].Asian Pac J Cancer Prev,2014,15(15):6181-6186.
[14] MOORE LD,LE T,FAN G.DNA methylation and its basic function[J].Neuropsychopharmacology,2013,38(1):23-38.
[15] XIAO FH,WANG HT,KONG QP.Dynamic DNA methylation during aging:A “prophet” of age-related outcomes[J].Front Genet,2019,10:107.
[16] CUELLO AC,HALL H,DO CARMO S.Experimental pharmacology in transgenic rodent models of Alzheimer’s disease[J].Front Pharmacol,2019,10:189.
[2] RICHARDSON A,HARIDASS SA,WARD E,et al.Investigation and treatment of premature ovarian insufficiency:A multi-disciplinary review of practice[J].Post Reprod Health,2018,2053369118811233.
[3] COULAM CB,ADAMSON SC,ANNEGERS JF.Incidence of premature ovarian failure[J].Obst Gynecol,1986,67(4):604-606.
[4] 钱星凯,夏杨柳,窦同意,等.儿茶酚氧位甲基转移酶与疾病的关系[J].药学学报,2016,51(4):543-551.
[5] GARNER EI,STOKES EE,BERKOWITZ RS,et al.Polymorphisms of the estrogen-metabolizing genes CYP17 and catechol-O-methyltransferase and risk of epithelial ovarian cancer[J].Cancer Res,2002,62(11):3058-3062.
[6] HILL LD,EWENS KG,MAHER BS,et al.Catechol-O-methyl-transferase (COMT) single nucleotide polymorphisms and haplotypes are not major risk factors for polycystic ovary syndrome[J].Mol Cell Endocrinol,2012,350(1):72-77.
[7] CORDTS EB,SANTOS MC,PELUSO C,et al.COMT polymorphism influences decrease of ovarian follicles and emerges as a predictive factor for premature ovarian insufficiency[J].J Ovarian Res,2014,7:47.
[8] QIN C,CHEN Y,LIN Q,et al.The significance of polymorphism and expression of oestrogen metabolism-related genes in Chinese women with premature ovarian insufficiency[J].Reprod Biomed Online,2017,35(5):609-615.
[9] MOUFARRIJ S,DANDAPANI M,ARTHOFER E,et al.Epigenetic therapy for ovarian cancer:Promise and progress[J].Clin Epigenetics,2019,11(1):7.
[10] KADER F,GHAI M,MAHARAJ L.The effects of DNA methylation on human psychology[J].Behav Brain Res,2018,346:47-65.
[11] HIRATA H,HINODA Y,OKAYAMA N,et al.COMT polymor-phisms affecting protein expression are risk factors for endometrial cancer[J].Mol Carcinog,2008,47 (10):768-774.
[12] DOHERTY JA,WEISS NS,FREEMAN RJ,et al.Genetic factors in catechol estrogen metabolism in relation to the risk of endometrial cancer[J].Cancer Epidemiol Biomarkers Prev,2005,14(2):357-366.
[13] LIU JX,LUO RC,LI R,et al.Lack of associations of the COMT Val158Met polymorphism with risk of endometrial and ovarian cancer:A pooled analysis of case-control studies[J].Asian Pac J Cancer Prev,2014,15(15):6181-6186.
[14] MOORE LD,LE T,FAN G.DNA methylation and its basic function[J].Neuropsychopharmacology,2013,38(1):23-38.
[15] XIAO FH,WANG HT,KONG QP.Dynamic DNA methylation during aging:A “prophet” of age-related outcomes[J].Front Genet,2019,10:107.
[16] CUELLO AC,HALL H,DO CARMO S.Experimental pharmacology in transgenic rodent models of Alzheimer’s disease[J].Front Pharmacol,2019,10:189.