吸烟对孕妇及胎儿GSTM1、GSTT1及CYP1A1基因多态性的影响

目的探讨吸烟对孕妇及胎儿谷胱甘肽S转移酶M1(GSTM1)、谷胱苷肽硫转移酶T1(GSTT1)及细胞色素P450(CYP1A1)基因多态性的影响。方法选取2016年1月至2017年6月在本院分娩的新生儿及母亲为研究对象,根据母亲是否吸烟分为主动吸烟组(22例)、被动吸烟组(289例)、不吸烟组(138例),采用PCR技术检测母亲及新生儿GSTM1、GSTT1及CYP1A1基因型。结果与不吸烟组比较,孕妇和新生儿GSTM1、GSTT1基因缺失型及CYP1A1基因突变型频率显著增加(P0.05),新生儿GSTM1、GSTT1基因缺失型及CYP1A1基因突变型与低出生体重、畸形等相关。结论吸烟增加孕妇和胎儿的GSTM1、GSTT1基因缺失型及CYP1A1基因突变型频率,并使胎儿低出生体重、畸形等发生率升高。     

中国汉族人群GSTT1 GSTM1基因多态性与血脂水平及冠心病的相关性研究

目的 通过研究中国汉族人群谷胱甘肽硫转移酶T1、M1(glutathione S-transferase,GSTT1、GSTM1)基因多态性与血脂水平及冠心病(coronary heart disease,CHD)之间的关系,探讨冠心病的遗传易感性.方法 采用病例-对照研究的方法,选取255例冠心病患者和与之年龄、性别相匹配的145例健康人作为对照组,采用聚合酶链式反应(polymerase chain reaction,PCR)方法检测GSTT1、GSTM1基因多态性,同时检测各研究对象的空腹血清总胆固醇(TC)、甘油三酯(TG)、载脂蛋白A(apoA)、载脂蛋白B(apoB)、高密度脂蛋白(HDL)、低密度脂蛋白(LDL)、脂蛋白a(Lpa)等临床资料.结果 冠心病组GSTT1、GSTM1、GSTT1/GSTM1阳性基因型频率低于对照组(P=0.007、P=0.003、P=0.005),GSTT1/GSTM1阴性基因型频率高于对照组(P=0.001).GSTM1阴性基因组apoB、LDL水平高于GSTM1阳性基因组(P=0.005、P=0.009);GSTT1/GSTM1阴性基因组apoB、LDL水平高于GSTT1/GSTM1阳性基因组(P=0.014、P=0.013).结论 GSTT1和GSTM1阴性基因型可能是中国汉族人群冠心病的遗传易感基因,GSTM1基因多态性可能通过影响血脂代谢而参与冠心病的发病过程.     

Bmi-1基因在多发性骨髓瘤中的研究进展

Bmi-1(B cell moloney murine leukemia virus insertion site 1,Bmi-1)基因是一种癌基因,属于PcG家族的一员.Bmi-1基因在维持正常造血干细胞(HSCs)和白血病干细胞(LSCs)的自我更新起重要作用,参与正常造血及血液恶性肿瘤的发生.研究证实,人类多种肿瘤的发生发展过程均伴有Bmi-1基因表达异常;同时,Bmi-1基因表达水平还可以作为部分肿瘤患者预后的指标之一,而高表达Bmi-1的肿瘤细胞可认为是“肿瘤干细胞(CSCs)”.目前研究发现,Bmi-1可在体内外调控多发性骨髓瘤(MM)细胞的生长、克隆,故其在此方面的研究日益受到重视.     

GSTM1、GSTT1基因多态性与白血病易感性的分析

目的分析GSTM1缺失基因型与白血病遗传易感性的关系。方法 131例白血病患者纳入病例组,200例体检健康者纳入对照组。采用巢式PCR检测GSTM1、GSTT1基因型。应用χ2检验和精确概率法比较各基因型频率在病例组与对照组之间的差异,用比值比(OR)及其95%的可信区间(CI)表示各基因型发生白血病的风险度。结果病例组GSTM1缺失基因型频率与对照组比较,差异有统计学意义(P=0.000 1,其OR值为2.152,95%CI为2.301~5.985);在病例组与对照组中均未检测到GSTT1缺失基因型;CML、AML与ALL组间进行GSTM1缺失基因型频率比较,差异无统计学意义(P0.05)。结论GSTM1缺失基因型可能是白血病的重要危险因素。     

骨髓增生异常综合征患者GSTT1、GSTM1基因分布频率和NQO1基因多态性分析

目的探讨谷胱甘肽S转移酶（GST）基因T1、M1（GSTY1、GSTM1）和苯醌氧化还原酶基因（NQO1C609T）多态性与骨髓增生异常综合征（MDS）易感性及MDS染色体核型异常的关系。方法用多重PCR方法检测52例MDS患者和241名与患者无血缘关系的正常人GSTF1和GSTM1基因型，用PCR-限制性片段长度多态性（RFLP）方法分析NQO1C609T基因型。结果与正常人对照组相比，MDS患者GSTT1和GSTM1无效型（nu11）比例明显增高（P值均〈0．01），其比值比（OR）分别为2．873（95％可信区间：1．491～5．537）和3．591（95％可信区间：1．717～7．508）。染色体核型正常的MDS患者GSTT1无效型比例较正常人对照组显著增高（OR=5．336，P〈0．01），而GSTM1无效型比例与正常人对照组比较差异无统计学意义染色体核型异常的MDS患者GSTM1无效型比例较正常人对照组显著增高（OR=3．740，P〈0．01），而GSTT1无效型比例与正常人对照组比较差异无统计学意义MDS患者的NQI1C609T各基因型与正常人对照组差异无统计学意义。结论GSTT1和GSTM1基因无效型可能与MDS发生相关，对判断MDS患者是否出现染色体核型异常有一定意义。     

沉默DNMT1基因对骨髓瘤细胞SOCS-1基因甲基化的影响

目的:探讨DNA甲基化转移酶1(DNA methyltransferase,DNMT1)基因沉默对多发性骨髓瘤RPMI8226细胞内细胞因子信号转导抑制分子1(suppressor of cytokine signaling,SOCS-1)甲基化的影响。方法:应用靶向沉默DNMT1基因的短发卡RNA(RNA-Short hairpin RNA,shRNA)通过脂质体(lipofectamineTM2000)介导转染骨髓瘤细胞RPMI8226,应用RT-PCR和Western blot方法分别检测转染前后DNMT1基因和蛋白表达,应用甲基化特异性PCR检测转染前后骨髓瘤细胞SOCS-1基因甲基化水平的变化。结果:转染后骨髓瘤细胞DNMT1 mRNA表达水平(0.176±0.004)和蛋白表达水平(0.065±0.014)均明显低于阴性对照组(0.956±0.033,0.415±0.027),其差异具有统计学意义(P0.01)。转染后SOCS-1基因甲基化水平较转染前明显降低。结论:利用shRNA能靶向沉默DNMT1基因的表达,并可以有效逆转SOCS-1基因高甲基化状态。     

GSTT1、GSTM1、NQO1、RAD51和XRCC3基因多态性与慢性粒细胞白血病发生关系的研究

目的探讨GSTT1、GSTM1、NQO1、RAD51和XRCC3基因多态性与我国慢性粒细胞白血病(CML)发生遗传易感之间的关系。方法共120例CML患者和458名与患者无血缘关系的正常人,用多重PCR方法检测GSTT1和GSTM1基因型,用PCR-RFLP方法分析RAD51,XRCC3,NQO1基因型。结果CML患者GSTT1和GSTM1缺失型比例分别为50.8%和59.2%,与正常对照组无显著差异(分别为42.8%和53.1%)。CML患者NQO1C/T和T/T基因型的比例(60.0%)、RAD51G135CG/C基因型比例(26.9%)和XRCC3-241Met杂合子缺失型(Thr/Met)的比例(9.2%)均与正常对照组(分别为65.3%,12.4%和9.2%)无统计学差异。结论本研究结果提示GSTT1、GSTM1、NQO1、RAD51和XRCC3基因型与我国CML的发生无显著相关性。     

SMPD1基因多态性与帕金森病发病关系的研究

目的分析SMPD1基因多态性与帕金森病(PD)发病的关系,为PD患者的治疗提供新的靶点。方法收集2017年6月至2019年1月该院神经内科收治的216例PD患者(PD组)和同期在本院体检的216名健康者(健康组)作为研究对象,参照SMPD1基因序列及相关文献设计引物序列,采用PCR和限制性内切酶酶切技术检测两组SMPD1基因多态性。结果 PD组rs1050228位点TT、CT、CC基因频率、等位基因T、C频率(分别为73.15%、5.56%、21.30%、76.07%、23.93%)与健康组(分别为72.22%、6.02%、21.76%、78.03%、21.97%)相比较差异均无统计学意义(P0.05);rs7951904位点的CC、CT基因频率、等位基因C、T频率(分别为99.07%、0.93%、99.64%、0.36%)与健康组(分别为99.54%、0.46%、99.81%、0.19%)相比较,差异均无统计学意义(P0.05);rs202081954位点的CC、CG基因频率、等位基因C、G频率(分别为99.07%、0.93%、99.64%、0.36%)与健康组(分别为99.54%、0.46%、99.81%、0.19%)相比较,差异均无统计学意义(P0.05);rs1050239位点的等位基因G、A频率(分别为81.43%、18.57%)与健康组(分别为91.62%、8.38%)相比较差异有统计学意义(P0.05);PD组和健康组均发现缬氨酸重复次数的多态,且PD组缬氨酸重复次数分布概率与健康组相比较,差异有统计学意义(P0.05)。结论 SMPD1基因突变型p.G508R及缬氨酸7个重复多态在PD的诊断或预防中可提供有用参考信息。     

Zo-1与id4基因异常甲基化在多发性骨髓瘤中的临床意义

多发性骨髓瘤(multiple myeloma,MM)是浆细胞恶性增殖性疾病,常规化疗不能治愈,为探讨zo-1、id4基因异常甲基化在MM诊断、预后、微小残留病检测及治疗中基因标志作用的临床意义,采用甲基化特异性聚合酶链反应(methylation specific PCR,MS-PCR)方法检测MM细胞系U266、H929及IM9(B淋巴母细胞,MM患者骨髓来源)中zo-1、id4基因的甲基化状况;应用MS-PCR方法分析20例MM患者及6例健康供者骨髓标本zo-1、id4基因启动子区甲基化状况。结果显示:zo-1、id4基因在MM细胞系中均为甲基化阳性(完全甲基化或部分甲基化);zo-1、id4基因在5例健康人标本中均呈完全非甲基化状态;MM患者骨髓中zo-1、id4基因甲基化阳性率分别是50%和85%,二者阳性覆盖率高达95%,二者均阳性的标本阳性率为40%,明显高于健康人(0%),差异均有统计学意义(p0.05);MM患者不同重链及不同轻链间甲基化阳性率没有统计学差异;MM患者是否具有B组症状并不影响患者的甲基化阳性率,这可能与标本数量有关。结论:MM患者中zo-1、id4基因甲基化状态发生改变并具有特异性,可能是MM新的基因标记。     

吸烟和CYP1A1、GSTM1基因多态对肺癌易感性影响

目的:探讨细胞色素P4501A1(cytochrome P-450 1A1,CYP1A1)与谷胱甘肤硫转移酶M1(glutathione S-transferase M1,GSTM1)基因多态与支气管肺癌的关系.方法:采用PCR限制性片段多态检测法,检测肺癌组103例和正常对照组138例患者的CYP1A1与GSTM1基因多态,以非条件性Logistic回归模型分别对年龄、性别进行校正后计算优势比及95%CI.结果:CYP1A1 ml突变型等位基因频率在对照组和肺癌组分别为27.6%和42.7%;GSTM1的缺陷型基因(D)频率在对照组、肺癌组分别为44.2%和61.2%.Logistic回归分析表明,CYP1A1(w1/m1)杂合型(B型)患肺癌的升高3.19倍,纯合突变型(C型)基因患肺癌的升高2.61倍,GSTM1缺陷型(D)患肺癌危险度升高2.09倍,差异均有统计学意义(P<0.05).GSTMl(D)/CYP1A1(B或C)基因型的个体患肺癌危险度为5.72倍.GSTMI(D)和CYP1A1 ml突变型等位基因均可明显增加鳞癌和小细胞癌的患病危险度.结论:CYP1A ml突变型等位基因和GSTM1(D)均是患肺癌的危险因素.CYP1A m1突变型等位基因和GSTM1(D)存在明显的交互作用,二者与吸烟有协同作用.     

GSTT1 and GSTM1 gene deletions are not associated with hepatotoxicity caused by antitubercular drugs

Background and objective: Susceptibility to antitubercular drug (ATD)‐induced hepatotoxicity may be genetically mediated, with variant alleles of genes such as N‐acetyltransferase (NAT2) and CYP2E1 reported as risk factors. Two studies of Asian populations have reported that GSTM1*0/*0 (null) genotype was a likely predictor of hepatotoxicity, whereas another of a Caucasian population implicated GSTT1*0/*0. We undertook a prospective case–control study to investigate whether GSTM*0/*0 and GSTT1*0/*0 were risk factors for ATD‐induced hepatotoxicity. Methods: Pulmonary tuberculosis patients on isoniazid, rifampicin and pyrazinamide who developed hepatotoxicity using defined criteria were prospectively identified. These cases were then matched with at least one control subject on the same drugs but without hepatotoxicity. Genotyping for GSTM1 and GSTT1 was performed by multiplex PCR on genomic DNA. The odds ratios for the frequency of specific GSTM1 and GSTT1 homozygotes in the case and control subjects were calculated to test for association between the genotypes and hepatotoxicity. Results and discussion: Hundred and fifty‐one subjects (51 cases, 100 controls) were enrolled. Odds ratio for GSTM1 null genotype was 1·00 (95% CI 0·51–1·97) and GSTT1 null was 2·02 (95% CI 0·39–10·39), respectively, showing that these genotypes are not associated with hepatotoxicity. Conclusion: GSTM1 *0/*0 or GSTT1 *0/*0 or both null genotypes, do not appear to be associated with ATD‐induced hepatotoxicity in our Indian population.     

CYP2E1, GSTM1 and GSTT1 genetic polymorphisms and susceptibility to antituberculosis drug-induced hepatotoxicity: a nested case-control study

What is Known and Objective: The pathogenic mechanism of antituberculosis drug‐induced hepatotoxicity (ATDH) is thought to involve drug‐metabolizing enzymes including N‐acetyl transferase2 (NAT2), cytochrome P4502E1 (CYP2E1) and glutathione S‐transferase (GST) M1, T1. The associations between genetic polymorphisms of those genes and ATDH have been reported but with inconsistent results. Moreover, most studies were hospital‐based retrospective studies and not prospective. We aimed to investigate possible associations of CYP2E1, GSTM1 and GSTT1 genetic polymorphisms with ATDH using a more robust case–control study nested in a population‐based prospective antituberculosis treatment cohort. Methods: A total of 4304 patients with smear‐positive tuberculosis (TB) who received standard short‐course chemotherapy were monitored for 6–9 months. Incidence density sampling method was adopted to select controls and 4 : 1 matched with each ATDH cases by age (±5 years), sex, treatment history, disease severity and drug dosage. The CYP2E1, GSTM1 and GSTT1 polymorphisms were genotyped using PCR–RFLP and multiplex PCR methods. Conditional logistic regression model was used to calculate odds ratio (OR) and 95% confidence interval (CI), as well as corresponding P‐values. Results and Discussion: A total of 89 ATDH cases and 356 controls were included in this study. There was no statistically significant association between CYP2E1 RsaI c1/c1 genotype or DraI C/C genotype and ATDH (OR = 0·99, 95% CI:0·62–1·59; OR = 1·13, 95% CI: 0·40–3·20, respectively) compared with CYP2E1 RsaI c1/c2 or c2/c2 genotypes or DraI D/D genotype, or between GSTM1/GSTT1 null genotypes and ATDH (OR = 1·22, 95% CI: 0·76–1·96; OR = 0·96, 95% CI: 0·60–1·52, respectively) compared with non‐null genotypes. What is new and Conclusion: This is the first study of the involvement of CYP2E1, GSTM1 and GSTT1 genetic polymorphisms in ATDH using a nested case–control population‐based prospective cohort design. We could not confirm positive associations of genetic polymorphisms of CYP2E1 RsaI, CYP2E1 DraI, GSTM1 null and GSTT1 null with ATDH reported by various groups, in our Chinese TB population.     

骨髓微环境对多发性骨髓瘤瘤细胞AP-1家族成员基因表达的调节作用

本研究旨在探讨骨髓微环境对多发性骨髓瘤(MM)细胞AP-1家族成员基因的调节作用。采用免疫磁珠方法分选8例多发性骨髓瘤患者CD138+的瘤细胞并与破骨细胞共培养,用实时定量PCR方法检测与破骨细胞共培养的瘤细胞和与破骨细胞共培养的瘤细胞AP-1家族成员c-jun、junD、fos和fosB的表达量。结果显示,多发性骨髓瘤患者的外周血单个核细胞经破骨细胞培养液培养后10-14天形成破骨细胞,瘤细胞与破骨细胞共培养后与未与破骨细胞共培养相比较,细胞活力明显增高,c-jun、junD、fos和fosB表达量均降低。结论:骨髓微环境可抑制AP-1家族成员的表达,促进MM瘤细胞存活,抑制瘤细胞凋亡。     

垂体瘤转化基因和C-myc基因在多发性骨髓瘤中表达的研究

本研究探讨垂体瘤转化基因（pituitary tumor transforming gene,阿TG）和c—myc基因在多发性骨髓瘤（multiplemyeloma,MM）患者中的表达并分析它们与MM发生的关系。采用RT—PCR方法检测33例MM患者及10例正常人骨髓单个核细胞（bone marrow mononuclear cell,BMMNC）中mG和c—myc基因的表达。结果表明：MM患者PTTG和c—myc基因的表达明显高于正常对照组（P〈0．05）。PTTG基因与c—myc基因的表达呈正相关（r=0．801,P〈0．05）。结论：mG和c—myc基因的过度表达与MM的发生发展有关。     

Xbp-1基因沉默对硼替佐米诱导人多发性骨髓瘤细胞凋亡的影响

本研究旨在探讨xbp-1基因沉默对硼替佐米诱导骨髓瘤细胞株NCI-H929(H929)凋亡的影响。通过小发夹RNA(shRNA)干扰xbp-1基因表达,采用Annexin V-FITC/PI双染流式细胞术检测靶细胞凋亡率,采用Wes-ternblot检测XBP-1蛋白水平。结果表明:通过慢病毒载体PLL3.7介导的xbp-1 shRNA表达,能有效抑制H929细胞XBP-1蛋白表达,xbp-1 shRNA表达细胞的凋亡率显著高于对照组[(10.13±0.61)%vs(2.5±0.2)%,p0.05];经硼替佐米处理后,xbp-1基因功能缺陷的H929细胞的凋亡率显著高于空载体组[(45.07±1)%vs(2.73±0.25)%,p0.05]。结论:xbp-1基因沉默能够显著增强硼替佐米对多发性骨髓瘤细胞的促凋亡活性。