Hedgehog信号通路基因Shh、Ptch1、Smo及Gli1在胰腺癌中的表达及意义

目的:检测Hedgehog信号通路基因Sonic Hedgehog(Shh)、Patched1(Ptch1)、Smoothened(Smo)及神经胶质瘤相关癌基因同源蛋白1(glioma-associated oncogene homolog 1,Gli1)在胰腺癌组织中的表达及其生物学意义.方法:采用反转录-聚合酶链反应(RT-PCR)分别检测48例胰腺癌组织和配对的癌旁组织中Shh、Ptch1、Smo及Gli1 mRNA的表达情况.结果:RT-PCR检测结果显示:Shh、Ptch1、Smo及Gli1 mRNA在胰腺癌组织中的相对表达量分别是0.652±0.036、0.604±0.063、0.493±0.011、0.512±0.052,在胰腺癌旁组织中为0.312±0.013、0.319±0.053、0.214±0.046、0.247±0.059(P0.05).与正常胰腺组织相比,胰腺癌组织中Shh、Ptch1、Smo及Gli1 mRNA表达量显著升高(P0.05),胰腺癌组织中Shh、Ptch1、Smo及G l i1mRNA表达与胰腺癌的分化程度有显著性差异(P0.05),与患者年龄、性别、肿瘤直径、TNM分期、淋巴结转移、糖链抗原(CA19-9)无显著性差异(P0.05).结论:Hedgehog信号通路基因Shh、Ptch1、S m o及G l i1在胰腺癌组织中表达增高,Hedgehog信号通路的异常激活可能与胰腺癌发生发展过程相关.     

Hedgehog信号通路相关蛋白在大鼠胰腺癌发生过程中的变化

目的 观察Hedgehog信号通路相关蛋白Ptch、Smo和Gli1在胰腺癌发生过程中的作用。方法 200只SD大鼠分成对照组(20只)、二甲基苯并蒽(DMBA)组(90只)和环巴明治疗组(90只)。通过胰腺被膜下直接置入DMBA方法诱导胰腺癌模型。环巴明组在置人DMBA后每天腹腔注射6.25 ml/kg 体重环巴明溶液。4个月后处死大鼠,观察胰腺组织病理学改变,应用免疫组化SP法检测胰腺癌及正常胰腺组织中Ptch、Smo、Gli1的蛋白表达。结果 DMBA组胰腺癌诱发成功率为57.5％ (46/80),诱发的肿瘤最大径为0.5～＞2.0 cm;环巴明组诱发成功率为17.1％ (14/82),肿瘤最大径为0.5 ～2.0 cm。两组胰腺癌发生率及肿瘤大小的差异具有统计学意义(P值均＜0.05)。DMBA组胰腺癌组织中Ptch、Smo及Gli1阳性表达率分别为82.6％、73.9％和65.2％;环巴明组分别为50.0％、42.9％和28.6％,两组差异具有统计学意义(P值均＜0.05)。3组的正常胰腺组织中均无Ptch、Smo、Gli1阳性表达。结论 较大剂量DMBA置入胰实质内可在短期内获得较高的胰腺癌发生率;胰腺癌组织Hedgehog信号蛋白表达显著增加;环巴明能抑制Hedgehog信号蛋白表达,进而抑制胰腺癌发生和发展。     

Hedgehog通路抑制剂逆转人胰腺癌SW1990细胞获得性耐药的实验研究

目的 探讨人胰腺癌SW1990耐药细胞株中Hedgehog通路成员(Shh、SMO、Gli1、ABCB1、ABCG2)表达的变化及应用Hedgehog信号通路抑制剂对其耐药性的影响.方法 建立人胰腺癌SW1990耐吉西他滨细胞株(SW1990/GZ).采用Hedgehog信号通路抑制剂环巴明作用耐药细胞,应用实时PCR、蛋白质印迹法检测用药前后细胞Shh、SMO、Gli1、ABCB1、ABCG2 mRNA和蛋白的表达及CD44+ CD24+、CD133+细胞亚群比例.以100 μmol/L吉西他滨作用环巴明处理后的SW1990/GZ细胞,应用Annexin V加PI双染法检测细胞凋亡率.结果 SW1990/GZ细胞中CD44+ CD24+细胞比例从亲代的(29.45±1.99)％增加到(93.16±2.46)％、CD133+细胞比例从(59.37±1.69)％增加到(95.88±1.47)％(P＜0.01);ABCB1、ABCG2、Shh、Gli1 mRNA的表达分别增加(274.90±31.44)、(3.48±0.33)、(12.07±1.71)、(4.15±0.42)倍(P＜0.01),并检测出SMO mRNA表达.蛋白表达结果与mRNA表达一致.2μmol/L环巴明作用SW1990/GZ细胞1周后,CD44+CD24+比例下降到(36.68±2.44)％、CD133+细胞比例下降到(62.76±1.28)％(P＜0.05).2、5μmol/L环巴明处理SW1990/GZ细胞后再用100 μmol/L吉西他滨处理细胞,细胞的凋亡加死亡率分别为(53.68±5.24)％和(69.99±3.16)％,显著高于对照组的(4.55±0.87)％(P＜0.01).结论 人胰腺癌SW1990耐药细胞株中高富集肿瘤干细胞,高表达Hedgehog信号通路成员SMO及Gli1.抑制耐药株的Hedgehog通路,可以降低其肿瘤干细胞比例,下调SMO及Gli1表达,部分恢复对吉西他滨的敏感性.     

RUNX3、 CyclinD1在胰腺癌组织中的表达及临床意义

目的 检测RUNX3、CyclinD1蛋白在胰腺癌组织中的表达,分析其临床意义。方法 采用免疫组化SP法检测47例胰腺癌、18例胰腺囊腺瘤及12例正常胰腺组织中RUNX3和CyclinD1蛋白的表达,分析它们与临床病理参数间的关系。结果 胰腺癌、胰腺囊腺瘤和正常胰腺组织的RUNX3蛋白阳性表达率分别为57.4％(27/47)、94.4％ (17/18)、100％( 12/12);CyclinDl蛋白阳性表达率分别为72.3％ (34/47)、44.4％(8/18)、8.3％(1/12)。胰腺癌RUNX3阳性表达与患者性别、年龄无关,与胰腺癌的临床分期、淋巴结转移、分化程度呈负相关(P＜0.05)。CyclinD1阳性表达与患者性别、年龄无关,与胰腺癌临床分期、淋巴结转移、分化程度呈正相关(P＜0.05)。胰腺癌组织RUNX3与CyclinDl的表达呈负相关(r= -0.375,P=0.009)。结论 胰腺癌组织中RUNX3呈低表达,Cyclin D1呈过表达,它们均与胰腺癌的发生、发展有关。     

Livin和Smac蛋白在胰腺癌中的表达及其意义

目的 探讨凋亡抑制蛋白Livin和促凋亡蛋白Smac在胰腺癌组织中的表达及其临床意义。方法 利用免疫组化SP法检测46例胰腺癌、15例胰岛素瘤和14例正常胰腺组织中Livin和Smac的表达,分析其与胰腺癌临床病理特征的关系。结果 胰腺癌、胰岛素瘤和正常胰腺组织Livin蛋白表达率分别为73.9％ (34/46)、73.3％ (11/15)和14.3％ (2/14)。胰腺癌、胰岛素瘤均高表达,两者间差异无统计学意义,但均显著高于正常胰腺组织(P值均＜0.05)。Livin蛋白的表达与胰腺癌淋巴结转移、组织病理分级及临床分期有关(P＜0.05或＜0.01)。胰腺癌、胰岛素瘤和正常胰腺组织Smac蛋白表达率分别为39.1％ (18/46)、100％( 15/15)和92.9％ (13/14)。正常胰腺和胰岛素瘤组织均高表达,两者间差异无统计学意义,但均显著高于胰腺癌组织(P值均＜0.05)。Smac蛋白的表达与胰腺癌淋巴结转移、组织病理分级、临床分期及患者年龄有关(P ＜0.05或＜0.01)。结论 Livin可能在胰腺癌的发生、发展中起重要促进作用,而Smac在防止胰腺癌的发生、发展中起一定的作用。     

Hedgehog信号通路在人肝细胞癌细胞系及肝癌组织中的表达与意义

目的探讨Hedgehog信号通路在人肝细胞癌细胞系及Shh基因在肝癌组织中的表达及意义。方法采用半定量RT-PCR法检测Hep3B和HCC-LM3细胞中Shh、Ihh、Ptch、Smo、Gli mRNA及21例肝细胞癌癌组织和癌旁肝组织中Shh mRNA的表达状况。结果Shh、Ihh、Ptch、Smo、Gli mRNA在两个肝癌细胞系中均有表达。其中,Shh、Gli1 mRNA在Hep3B细胞中的表达强度显著高于HCC-LM3细胞,而Gli2和Gli3的表达强度正相反;Ihh、Smo mRNA在Hep3B细胞中的表达强度高于HCC-LM3细胞,但两者间无显著性差异;Ptch mRNA在两个细胞系细胞中的表达无显著性差异;Shh mRNA在肝细胞癌癌组织中的阳性检出率为57.1%（12/21）,在癌旁组织中为4.8%（1/21）,两者间有显著性差异（P〈0.05）。其中,6例高分化、8例中分化和7例低分化肝细胞癌中Shh mRNA的阳性检出率分别为83.3%（5/6）、62.5%（5/8）和28.6%（2/7）,表现为随肝细胞癌由高到低的分化而呈显著性下降的趋势（P〈0.05）;Shh mRNA的表达强度在肝细胞癌癌组织之间、癌与癌旁组织之间无显著性差异。结论Hedgehog信号通路的异常激活参与了肝细胞癌的发生发展过程,Shh mRNA在部分高中分化肝细胞癌中表达上调,这或许为探讨肝癌的发生机制、早期诊断及治疗提供了新的依据。     

内分泌腺来源的血管内皮细胞生长因子在人胰腺癌组织中的表达及临床意义

目的 探讨内分泌腺来源的血管内皮细胞生长因子(EG-VEGF)在人胰腺癌组织中的表达及临床意义.方法 应用免疫组化技术检测EG-VEGF在60例胰腺癌组织和10例正常胰腺组织中的表达情况,并分析其与临床病理因素之间的关系.结果 EG-VEGF在60例胰腺癌组织中的阳性表达率为95％(57/60),而在10例正常胰腺组织中的阳性表达率为10％( 1/10),两者间差异有统计学意义(P＜0.05).EG-VEGF的表达与胰腺癌的临床病理分期以及淋巴结转移呈正相关(P＜0.05),而与性别、年龄、病理分级无明显相关性(P＞0.05).结论 EG-VEGF在胰腺癌的发生发展过程中起着重要作用,可作为判断胰腺癌预后的有用指标.     

DLL4在胰腺癌组织中的表达及其与肿瘤血管生成的关系

目的 检测delta-like ligand 4(DLL4)在胰腺癌中的表达,分析其与肿瘤血管生成及临床病理特征的关系.方法 采用免疫组化SP法检测60例胰腺癌组织及20例癌旁正常胰腺组织中DLL4的表达;观察微血管内皮细胞CD34表达,计算微血管密度(MVD);分析DLL4表达、MVD与胰腺癌临床病理特征之间的关系以及两者的相关性.结果 DLL4在胰腺癌组织中的表达明显高于正常胰腺组织(68.3％比20.0％,P＜0.01);胰腺癌组织DLL4的高表达与肿瘤分化程度、TNM分型、肿瘤转移及浸润等呈正相关(P值均＜0.05),而与肿瘤部位、大小、组织病理分型无关.胰腺癌组织MVD明显高于正常胰腺组织(34.9±13.2比18.9±2.2,P＜0.01);胰腺癌的MVD与肿瘤分化程度、TNM分期、肿瘤转移及浸润有关(P值均＜0.05),而与肿瘤部位、大小、组织病理分型等无明显相关性.DLL4表达阳性的胰腺癌组织MVD明显高于DLL4表达阴性者(38.8±10.7比29.0±15.2,P＜0.05),且DLL4表达与MVD呈显著正相关(r=0.669,P＜0.05).结论 DLL4表达可促进肿瘤血管生成,且与胰腺癌的转移、浸润相关.     

载脂蛋白E和补体C4b1在人胰腺癌组织中的表达及意义

目的 检测载脂蛋白E(ApoE)和补体C4b1在人胰腺癌组织中的表达,探讨其意义.方法 应用免疫组化法检测38例胰腺癌和相应癌旁正常胰腺组织ApoE、C4b1蛋白表达;应用RT-PCR法检测20例胰腺癌和相应癌旁正常胰腺组织ApoE、C4b1 mRNA表达.分析ApoE、C4b1的表达与胰腺癌生物特征的相关性.结果 胰腺癌组织ApoE、C4b1蛋白表达阳性率分别为86.8%(33/38)和76.3%(29/38),显著高于癌旁正常胰腺组织的42.1%(16/38)和26.3%(10/38)(P值均＜0.01);有淋巴结转移的胰腺癌组织ApoE、C4b1蛋白表达阳性率分别为78.3%(18/23)和73.9%(17/23),明显高于无转移者的33.3%(5/15)和40.0%(6/15)(P值均＜0.05).胰腺癌ApoE、C4b1 mRNA表达量分别为4.83±0.65、7.94±0.95,明显高于癌旁正常胰腺组织的1.78±0.74、1.22±0.57(P值均＜0.01);有淋巴结转移的胰腺癌组织ApoE、C4b1 mRNA的表达量为5.05±0.71、8.24±1.07,显著高于无转移者的4.42±0.25、7.39±0.15(P值均＜0.05).结论 ApoE、C4b1在胰腺癌组织中高表达,并与胰腺癌淋巴结转移相关.     

miR-135b、LZTS1、β-catenin在胰腺癌中的表达及临床意义

目的:探讨miR-135b、LZTS1及β-catenin在胰腺癌中的表达意义及三者之间的相关性.方法:分别采用锁定核酸原位杂交技术(LNA-ISH)和免疫组织化学法检测miR-135b、LZTS1和β-catenin蛋白在70例胰腺癌及相应癌旁组织中的表达情况.结果:miR-135b在胰腺癌中的表达率高于癌旁正常组织(71.4%vs 42.9%,P=0.001),LZTS1、β-catenin蛋白在胰腺癌中的表达率均低于癌旁正常组织(34.3%vs 68.6%、34.3%vs 74.3%,P0.05).在胰腺癌组织中miR-135b与LZTS1的表达水平呈负相关(r=-0.61,P0.05),与β-catenin无相关性(r=0.06,P0.05);LZTS1与β-catenin呈正相关(r=0.37,P0.05),且miR-135b、LZTS1和β-catenin的阳性表达水平与胰腺癌的临床分期及淋巴结转移密切相关(P0.05),与胰腺癌患者年龄、性别、肿瘤部位及组织学分级无关(P0.05).结论:miR-135b在胰腺癌中表达升高,LZTS1与β-catenin在胰腺癌中表达减少,并且表达升高的miR135b可能通过下调LZTS1基因表达而参与了胰腺癌的发生发展.     

Characterization of MTHFR,GSTM1, GSTT1, GSTP1, and CYP1A1 genotypes in childhood acute leukemia

The role of methylenetetrahydrofolate reductase (MTHFR C677T), glutathione S-transferases (GSTM1 and GSTT1 null, GSTP1 Ile105Val), and cytochromes p450 (CYP1A1*2A) genotypes in the etiology of childhood leukemia was simultaneously investigated. 144 Turkish children with acute lymphoblastic leukemia (ALL) and 33 with acute nonlymphoblastic leukemia (ANLL) were studied and compared with 185 healthy pediatric controls. The frequency of MTHFR genotype was insignificantly higher in ALL (7.7%) and ANLL (6.3%) than in controls (4.4%). Equal distribution of the GSTM1 null genotype was detected between ALL patients and controls (55%), while its incidence was slightly higher in ANLL patients (61.3%). Although GSTT1 null genotype was insignificantly lower in ALL patients (20.9%) than controls (22.7%), it was significantly underrepresented in ANLL patients (6.5%) (P = 0.05, OR 0.24, 95% CI 0.05-1.03). The homozygous frequency of GSTP1 genotype did not differ significantly between groups of ALL (3.7%), ANLL patients (9.1%) and controls (4.9%). Homozygous CYP1A1*2A genotype was underrepresented in ALL patients (1%) as compared to control (4.8%) but the differences did not reach to statistical significance (OR 0.21; 95% CI 0.03-1.72). Homozygosity for this genotype was not detected in ANLL patients. No particular association was noted between different combinations of combined genotypes and risk of development of childhood ALL and ANLL. These results suggested that there are no significant associations between the studied genotypes and the risk of developing either form of acute leukemia except GSTT1 null and homozygosity for CYP1A1 genotypes that may play protective roles in the development of ANLL in Turkish children.     

PICCO指导重症急性胰腺炎早期液体复苏的效果评价

目的 探讨脉搏连续心排血量（PICCO）指导早期液体复苏对重症急性胰腺炎（SAP）的临床意义。方法 选择我院消化科自2013年1月～2015年1月收治并应用PICCO指导早期液体复苏的37例SAP患 者作为PICCO组，同期选择应用中心静脉压（CVP）指导液体复苏的39例SAP患者作为对照组，比较两组48h内液体出入量、血管活性药物使用时间，以及机械通气时间、ICU住院时间和28天病死率。并应用受试者工作特征性（ROC）曲线分析28天病死率的危险因素。结果 共有76例患者入选，其中男41例，年龄58.76±13.84岁。两组间年龄、性别比例、入院血糖、血乳酸、血肌酐、氧和指数、平均动脉压、APACHE II 评分均无显著差异（P>0.05）。PiCCO组患者的0～6h补液量明显多于对照组（P<0.05），而6～72h补液量较对照组明显减少（P＜0.05）。PICCO组患者的血液净化率、机械通气时间、ICU住院时间均显著减少（P<0.05），但是两组患者应用血管活性药物的比例、导管相关感染率和28天病死率均无显著差别（P>0.05）。ROC曲线发现年龄（AUC 0.71, 95% CI, 0.63～0.76，P=0.03）和APACHE II评分（AUC 0.78, 95% CI, 0.67～0.91，P=0.02）为预测28天病死率的重要因素。结论PiCCO可以精确指导SAP患者早期液体复苏，并减少机械通气时间和ICU住院时间。     

The polymorphisms of GSTM1, GSTT1, HYL1*2, and XRCC1, and aflatoxin B1-related hepatocellular carcinoma in Guangxi population, China.

AIM: High incidence rates of hepatocellular carcinoma (HCC) in Guangxi, China, are primarily due to heavy aflatoxin B1 (AFB1) exposure via corn and groundnut consumption. This study was designed to examine the polymorphisms associated of three carcinogen-metabolizing genes (namely: GSTM1, GSTT1, and HYL1*2) and one DNA-repair gene (namely: XRCC1), and investigate their role as susceptibility markers for HCC. METHODS: We conducted a case-control study including 257 cases of cancer and 649 hospital-based age, sex, ethnicity, and hepatitis B virus infection-matched controls to examine the role of genetic polymorphisms of four genes (GSTM1, GSTT1, HYL1*2, and XRCC1) in the context of HCC risk for the Guangxi population. Genomic DNA isolated from 2ml whole blood was used to genotype GSTM1, GSTT1, HYL1*2, and XRCC1 by means of polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis. RESULTS: GSTT1-null genotype was not significantly associated with the risk of HCC, but GSTM1-null genotype [adjusted odds ratio (OR)=2.29, 95% confidence interval (CI)=1.59-3.31], HYL1*2 genotypes with 113 His allele (namely: YH/HH, adjusted OR=2.55, CI=1.78-3.65), and XRCC1 genotypes with 399 Gln allele (namely: AG/GG, adjusted OR=2.47, CI=1.72-3.54) increased the HCC risk. Compared with those individuals who did not express any putative risk genotypes as reference (OR=1), individuals featuring all of the putative risk genotypes [GSTM1-null, HYL1*2-YH/HH, and XRCC1-AG/GG] did experience a significantly greater cancer risk (adjusted OR=10.83, CI=5.44-21.59, P(interaction)<0.01). Additionally, the risk of HCC did appear to differ more significantly among individuals featuring risk genotypes and high-level or long-term AFB1 exposure, whose adjusted ORs (CIs) were 52.44 (17.51-157.08) and 326.93 (38.58-2770.52), respectively. CONCLUSIONS: The results suggest that carcinogen metabolism and DNA-repair pathways may simultaneously modulate the risk of HCC for Guangxi population, and, particularly for these having high-level or long-term AFB1 exposure.     

MChip, a low density microarray, differentiates among seasonal human H1N1, North American swine H1N1, and the 2009 pandemic H1N1

Please cite this paper as: Heil et al. (2010) MChip, a low density microarray, differentiates among seasonal human H1N1, North American swine H1N1, and the 2009 pandemic H1N1. Influenza and Other Respiratory Viruses 4(6), 411–416. Background  The MChip uses data from the hybridization of amplified viral RNA to 15 distinct oligonucleotides that target the influenza A matrix (M) gene segment. An artificial neural network (ANN) automates the interpretation of subtle differences in fluorescence intensity patterns from the microarray. The complete process from clinical specimen to identification including amplification of viral RNA can be completed in <8 hours for under US$10. Objectives  The work presented here represents an effort to expand and test the capabilities of the MChip to differentiate influenza A/H1N1 of various species origin. Methods  The MChip ANN was trained to recognize fluorescence image patterns of a variety of known influenza A viruses, including examples of human H1N1, human H3N2, swine H1N1, 2009 pandemic influenza A H1N1, and a wide variety of avian, equine, canine, and swine influenza viruses. Robustness of the MChip ANN was evaluated using 296 blinded isolates. Results  Training of the ANN was expanded by the addition of 71 well‐characterized influenza A isolates and yielded relatively high accuracy (little misclassification) in distinguishing unique H1N1 strains: nine human A/H1N1 (88·9% correct), 35 human A/H3N2 (97·1% correct), 31 North American swine A/H1N1 (80·6% correct), 14 2009 pandemic A/H1N1 (87·7% correct), and 23 negative samples (91·3% correct). Genetic diversity among the swine H1N1 isolates may have contributed to the lower success rate for these viruses. Conclusions  The current study demonstrates the MChip has the capability to differentiate the genetic variations among influenza viruses with appropriate ANN training. Further selective enrichment of the ANN will improve its ability to rapidly and reliably characterize influenza viruses of unknown origin.     

CYP1A1 , GSTM1 , GSTT1 and NQO1 polymorphisms and colorectal adenomas in Japanese men

AIM: To investigate the role of functional genetic poly-morphisms of metabolic enzymes of tobacco carcinogens in the development of colorectal adenomas. METHODS: The study subjects were 455 patients with colorectal adenomas and 1052 controls with no polyps who underwent total colonoscopy in a preretirement health examination at two Self Defense Forces hospitals. The genetic polymorphisms studied wereCYP1A1*2A (rs 4646903), CYP1A1*2C (rs 1048943), GSTM1 (null or non-null genotype), GSTT1 (null or non-null genotype) and NQO1 C609T (rs 1800566). Genotypes were determined by the polymerase chain reaction (PCR)-restriction fragment length polymorphism or PCR method using genomic DNA extracted from the buffy coat. Cigarette smoking and other life-style factors were ascertained by a self-administered questionnaire. The associations of the polymorphisms with colorectal adenomas were examined by means of OR and 95%CI, which were derived from logistic regression analysis. Statistical adjustment was made for smoking, alcohol use, body mass index and other factors. The gene-gene interaction and effect modification of smoking were evaluated by the likelihood ratio test. RESULTS: None of the five polymorphisms showed a significant association with colorectal adenomas, nor was the combination of GSTM1 and GSTT1 . A borderline significant interaction was observed for the combination of CYP1A1*2C and NQO1 (P = 0.051). The OR associated with CYP1A1*2C was significantly lower than unity among individuals with the NQO1 609CC genotype. The adjusted OR for the combination of the CYP1A1*2C allele and NQO1 609CC genotype was 0.61 (95%CI: 0.42-0.91). Although the interaction was not statistically significant (P = 0.24), the OR for individuals carrying the CYP1A1*2C allele and GSTT1 null genotype decreased significantly compared with those who had neither CYP1A1*2C allele nor GSTT1 null genotype (adjusted OR: 0.69, 95%CI: 0.49-0.97). Smoking did not modify the associations of the individual polymorphisms with colorectal adenomas. There w     

Analysis of HLA-DRB1, DQA1, DQB1 haplotypes in Sardinian centenarians

Some genetic determinants of longevity might reside in those polymorphisms for the immune system genes that regulate immune responses. Many longevity association studies focused their attention on HLA (the human MHC) polymorphisms, but discordant results have been obtained. Sardinians are a relatively isolate population and represent a suitable population for association studies. Some HLA-DR and DQ alleles form very stable haplotypes with a strong linkage disequilibrium. In a previous study on Sardinian centenarians we have suggested that HLA-DRB1 *15 allele might be marginally associated to longevity. HLA-DR,DQ haplotypes are in strong linkage disequilibrium and well conserved playing a role in the association to diseases. Hence, we have evaluated, by amplification refractory mutation system/polymerase chain reaction (ARMS-PCR) the HLADQA1 and HLA-DQB1 allele frequencies in 123 centenarians and 92 controls from Sardinia to assess whether the association to HLA-DRB1 *15 allele may be due to the other genes involved in the HLA-DR,DQ haplotypes. The frequencies of HLA-DQA1, DQB1 haplotypes were not significantly modified in centenarians. Nevertheless by evaluating the frequency of DRB1 *15 linked haplotypes, we observed a not significant increase in centenarians of HLA-DQA1 *01, DQB1 *05 and HLA-DQA1 *01,DQB1 *06 haplotypes. These data suggest that these haplotypes might have a role in determining life span expectancy and longevity.     

Association analyses of genetic polymorphisms of GSTM1, GSTT1, NQO1, NAT2, LPL, PRSS1, PSTI, and CFTR with chronic alcoholic pancreatitis in Japan

Background: Excessive consumption of alcohol is involved in the onset of pancreatitis. However, most of heavy drinkers do not always develop chronic pancreatitis. Various genetic factors appear to be involved in these individual differences in onset of chronic alcoholic pancreatitis. Here we investigated a possible association of alcoholic pancreatitis with polymorphisms of the various genes belong to the phase II detoxification enzymes responsible for metabolism of the oxidative compounds, and the several genes that have relevance to inherited pancreatitis. Methods: The subjects consisted of 53 patients with chronic alcoholic pancreatitis, 54 alcoholic patients without pancreatic dysfunction, and 42 healthy individuals. DNA was extracted from the peripheral nucleated blood cells of all subjects and genetic mutations and subtypes were analyzed by the PCR and RFLP methods. We examined the correlation between chronic alcoholic pancreatitis and variants of the phase II detoxification enzymes such as Glutathione S-transferase M1 (GSTM1), glutathione S-transferase theta 1 (GSTT1), NADPH-quinone oxidoreductase 1 (NQO1), and N-acetyl transferase (NAT2). In addition, genes of lipoprotein lipase (LPL), cationic trypsinogen (PRSS1), pancreatic secretory trypsin inhibitor (PSTI), and cystic fibrosis transmembrane conductance regulator (CFTR) were also analyzed. Results: Frequencies of the gene deletion of GSTM1 and GSTT1 in addition to the C-allele frequency of NQO1 tended to be higher in the alcoholic patients with (AlCP) or without pancreatic dysfunction (Alc) than in the healthy controls although the difference was not significant. The NAT2 gene showed no relation with Alc and AlCP patients. PSTI, LPL, PRSS1, and CFTR genes presented no association with chronic alcoholic pancreatitis. Conclusions: All genes analyzed in the present study lacked association with chronic alcoholic pancreatitis. However, the gene deletion of GSTM1 and GSTT1, and the C-allele of NQO1 cannot be ruled out for association with alcoholism.     

Role of CYP2D6, CYP1A1, CYP2E1, GSTT1, and GSTM1 genes in the susceptibility to acute leukemias

Acute leukemias (ALs) are heterogeneous diseases. Functional polymorphisms in the genes encoding detoxification enzymes cause inter-individual differences, which contribute to leukemia susceptibility. The CYP2D6, CYP1A1, CYP2E1, GSTT1, and GSTM1 polymorphisms in ALL (n = 156) and AML (n = 94) patients and 140 healthy controls were genotyped by PCR and/or PCR-RFLP using blood or bone marrow samples. No association was observed between the GSTT1 gene deletion and patients (OR = 0.8, 95% CI = 0.4-1.7 for AMLs and OR = 0.9, 95% CI = 0.5-1.6 for ALLs). Patients with ALL and AML had a higher prevalence of the GSTM1 deletions compared to controls but only the difference among adult AML patients (OR = 2.1, 95% CI = 1.0-4.2) was statistically significant. The CYP2D6*3 variant allele frequency was lower in the overall acute leukemia patients (0.6%) compared to controls (P = 0.03). CYP2D6*1/*3 genotype frequency also showed a protective association in AML patients (OR = 0.09, 95% CI = 0.01-1.7; P = 0.04). We also found a risk association for CYP2E1*5 in ALL and AML (OR = 3.6, 95% CI = 1.4-9.4 and OR = 3.9, 95% CI = 1.4-10.5, respectively). No association was found for the studied CYP2D6*4, CYP1A1*2A, and GSTT1"null" variants and the risk of acute leuke-mia (ALL or AML). This case-control study suggests a contribution of CYP2E1, CYP2D6, and GSTM1 "null" variants to the development of acute leukemias.     

CYP1A1, CYP2E1, GSTM1, GSTT1, EPHX1 exons 3 and 4, and NAT2 polymorphisms, smoking, consumption of alcohol and fruit and vegetables and risk of head and neck cancer

Purpose As risk-modifiers of alcohol and tobacco effects, metabolic genes polymorphisms were investigated as susceptibility candidates for squamous cell carcinoma of the head and neck (SCCHN). Methods A total of 210 cases and 245 hospital controls, age and gender matched, were genotyped for CYP1A1, CYP2E1, GSTM1, GSTT1, EPHX1 exons 3 and 4, and NAT2 polymorphisms. A measurement of the biological interaction among two risk factors was estimated by the attributable proportion (AP) due to interaction and its 95% confidence interval (CI). Results SCCHN risk was associated with high-levels of alcohol intake [OR = 3.50 (95%CI: 1.93–6.35) and OR = 6.47 (95%CI: 2.92–14.35) for 19–30 g/day and >30 g/day, respectively], cigarette smoking [OR = 3.47 (95%CI: 1.88–6.41) and OR = 7.65 (95%CI: 4.20–13.90) for 1–25 and >25 pack-years of smoking, respectively] and low-fruit and vegetables consumption (OR = 2.45; 95%CI: 1.53–3.92). No differences were observed for the genotypes or haplotypes distributions among cases and controls, and no biological interaction emerged from gene–gene and gene–environment interaction analyses. An attributable proportion (AP) due to biological interaction of 0.65 (95%CI: 0.40–0.90) was detected for heavy drinkers with a low intake of fruit and vegetables, and an AP of 0.40 (95%CI: 0.10–0.72) resulted forever smokers with low fruit and vegetables consumption. Conclusions Even in presence of high alcohol consumption or cigarette smoking, a high intake of fruit and vegetables might prevent the development of around one quarter of SCCHN cases. The lack of interaction between the studied polymorphisms and the environmental exposures suggests that chronic consumption of tobacco and alcohol overwhelm enzyme defences, irrespective of genotype.