健康更年期后女性AR基因(CAG)n多态性的调查

目的 研究健康更年期后女性雄激素受体基因(CAG)n重复序列多态性.方法 使用DNA测序方法 对60例更年期后健康女性AR基因外显子1 NH2端转录调节区内(CAG)n重复序列长度进行测定.结果 (1)健康更年期后女性AR基因(CAG)n重复数最短为18,最长为34,平均值为23.34±2.44;(2)更年期后女性(CAG)n重复序列多态性与育龄妇女有差异(P＞0.05).结论(1)健康更年期后女性AR基因(CAG)n重复序列长度存在多态性;(2)健康更年期后女性AR基因(CAG)n的重复数大于育龄期女性.     

健康老年男性雄激素受体基因CAG重复序列多态性研究

目的研究健康老年男性雄激素受体(AR)基因CAG重复序列多态性。方法使用DNA测序方法对102例健康老年男性AR基因外显子1 NH_2-端转录调节区内CAG重复序列长度进行测定。结果男性AR基因CAG重复数最短为8,最长为35。CAG重复数平均值为22．83±3．97。CAG重复数分布频率最多的为21(17．6%),22 (14．7%),23(15．7%)和24(6．9%),共占54．9%。结论健康老年男性AR基因CAG重复序列呈现多态性,多数集中在21-24。为进一步研究AR基因变异与疾病的关系提供依据。     

雄激素受体基因(CAG)n多态性与男性心血管病危险因素

众所周知，年龄、性别、家族史、吸烟、高血压、血脂异常、糖尿病、体力活动减少、肥胖和精神压力等为冠心病的十大危险因素。男性性别已成为心血管疾病的独立危险因子。目前，雄激素与男性心血管疾病的关系并不十分明确，而体内雄激素是通过雄激素受体（AR）发挥生理作用。AR基因转录活性的细微调节已被发现在AR NH2-端由多聚谷氨酸（Gln）n的可变长度决定，并由其外显子1三核苷酸（CAG）n重复序列编码。     

动脉粥样硬化遗传易感性与HUMARA（GGC）n基因多态性的相关性研究

目的 :研究雄激素受体基因 （HU MARA ）多态性在陕西地区老年动脉粥样硬化 （AS）人群中分布规律及其该位点与 AS遗传易感性的关系。方法 :应用 PCR- Am p- FL P（多聚酶链反应扩增片段长度多态性 ）技术对老年 AS患者 10 0例及非 AS老年人 78例 HU MARA - STR（短串联重复序列 short tandem repeats）位点进行多态性分析。结果 :在正常人群中 HU MARA基因 GGC位点共检测出 16种基因型 ,13个等位基因 ,其片段大小在 177～ 2 6 7bp,杂合度为 85 %,多态信息量 （polymorphism information content,PIC）为 0 .80 ;AS人群 HU MARA基因 GGC位点共检出 15种基因型 ,13个等位基因 ,其片段大小为 195～ 2 37bp,杂合度为 77%,多态信息量为 0 .72 ,两群体中基因频率分布有显著差异 （P<0 .0 1） ,发现某些片段存在基因频率分布高峰 ,在两群体中亦有差异性。结论 :推测 HU -MARA基因 STR多态位点与 AS的遗传易感性相关。     

雄激素受体基因(CAG)n多态性与绝经后骨质疏松关系的研究

目的 探讨雄激素受体(AR)基因第1外显子三核苷酸胞嘧啶-腺嘌呤-鸟嘌呤基因重复序列[(CAG)n]多态性在绝经后骨质疏松(PMO)发生中的作用.方法 采用美国NORLAND公司的XR-46系列双能X线骨密度仪测量股骨颈及腰椎2～4(L2～4)骨密度,根据股骨颈和L2～4骨密度诊断为PMO分别为78例和108例(PMO组),非PMO分别为73例和60例(对照组).采用基因扫描及DNA测序技术检测AR基因(CAG),重复次数和基因型,探讨(CAG)n多态性与PMO的关系.结果 检测到(GAG)n多态性的18、20、21、23、24、25、26、27、28、29及30次重复11个等位基因及16种基因型.基因型和等位基因频率分布在根据股骨颈骨密度诊断的PMO组(SS:25.6%,SL:39.7%,LL:34.6%;S:45.5%,L:54.5%)与对照组(SS:23.3%,SL:45.2%,LL:31.5%;S:45.9%,L:54.1%)比较,差异无统计学意义(均为P＞0.05);SL基因型(OR=0.798,95%CI:0.335～1.797)、LL基因型(OR=0.998,95% CI:0.425～2.341)或SL+LL基因型(OR=0.880,95%CI:0.419～1.852)者与SS型者比较,患PMO风险差异无统计学意义(均为P＞0.05).L2～4骨密度诊断的PMO组的基因型和等位基因频率分布(SS:18.5%,SL:49.1%,LL:32.4%;S:43.1%,L:56.9%)与对照组(SS:21.7%,SL:45.0%,LL:33.3%;S:44.2%,L:55.8%)比较,差异无统计学意义(均为P＞0.05);SL基因型(OR=1.276,95%CI:0.552～2.950)、LL基因型(OR=1.137,95%CI:0.468～2.766)或SL+LL基因型(OR=1.217,95%CI:0.556～2.663)者与SS型者比较,患PMO风险差异亦无统计学意义(均为P＞0.05).调整年龄、绝经时间、绝经年龄及体质指数后,Logistic回归分析显示,(CAG)n多态性与股骨颈及L2～4骨密度诊断的PMO均无相关性(P＞0.05).结论 AR基因(CAG)n多态性可能与PMO的发生无关.     

低密度脂蛋白受体基因二核苷酸串联重复序列(TA)n在正常中国汉族人群的多态性分布

目的　研究 LDL- R基因二核基酸串联重复序列 (TA) n在正常中国汉族人群的多态分布。方法　应用 PCR扩增 (TA) n所在区域的 DNA,通过变性聚丙烯酰胺凝胶电泳 ,结合银染技术检测扩增产物 ,测定了 2 0 6例正常中国汉族人的等位基因片段。结果　检测到 5种不同长度的等位基因片段和 1 0种不同基因型 ,多态信息容量 (PIC)为 0 .51 97,杂合度 0 .52 0 9。结论　该位点在中国人群中具有长度多态性 ,且其多态性分布与西方人群存在明显差异。     

多巴胺转运体基因可变数目串联重复多态在帕金森病易感性中的作用

目的　探讨多巴胺转运体 (DopamineTransporter ,DAT)基因可变数目串联重复多态(variablenumberoftandemrepeats,VNTR)在帕金森病的疾病易感性中的作用。　方法　采用扩增片段长度多态法 (Amplifiedfragmentlengthpolymorphism ,Amp FLP) ,在上海汉族人群中选择 14 4例帕金森病患者和 184例健康人 ,进行DAT基因VNTR多态与帕金森病的遗传关联研究。　结果　(1)上海地区汉族人群中 ,DAT基因VNTR多态以 4 80bp重复片段为主 ,其基因频率为 93% ;(2 )帕金森病患者与健康人群间无DAT基因VNTR多态分布的差异 ,该多态与帕金森病亦无关联 (P >0 0 5 )。按发病年龄中位数为界将帕金森病组分层后 ,无论是 <6 0岁组人群 ,还是≥ 6 0岁组人群 ,都不存在该多态性与帕金森病的相关。　结论　上海地区汉族人群中DAT基因多态性与帕金森病无关。     

雄激素受体基因多态性与男性冠心病关系的研究

目的:雄激素受体(AR)基因外显子1区基因片断CAG多态性与男性冠心病(CAD)及其危险因素的关系。方法:125例冠脉造影证实的男性冠心病人均接受检查,CAD严重程度采用冠脉狭窄≥50%的支数(0～3),分别计算心肌梗塞栓塞(TIMI)危险积分,外周血AR转录激活区CAG重复序列的长度采用PCR法测定。结果:CAG重复长度为13～30次,CAG重复≥24次者为长AR基因组,<24次者为短AR基因组。与长AR基因组比较:短AR基因组患者具有较低的平均体重指数(BMI)(t=-3.024,P=0.005)和较低水平的高密度脂蛋白(HDL)(t=-2.610,P=0.010);短AR基因组出现明显冠脉狭窄的可能性较大(x2=4.82,P=0.028),TIMI危险积分较高(M-W检验,z=-2.644,P=0.008)。Spearman相关分析表明,AR基因CAG长度和TIMI危险积分显著负相关(r=-0.230,P=0.010),这种相关性独立于年龄、BMI、LDL、UA和HDL之外(偏相关系数r=-0.250,P=0.006)。结论:具有短AR基因者与更加严重的冠心病相关,提示CAG多态性与男性CAD病情发展相关,AR表达增多,雄激素敏感性增高可能是男性CAD发病率较高的原因之一。     

MMP-9基因二核苷酸重复序列多态标志与2型糖尿病视网膜并发症相关

采用基因扫描方法 ,检测 2型糖尿病视网膜并发症患者 (DR )及无该并发症 (DR -)患者基质金属蛋白酶 9(MMP 9)基因二核苷酸串联重复序列多态标记 (AC)n的基因型。与DR -组相比 ,DR 组AC2 1基因型及等位基因频率显著减少 ,提示MMP 9基因的AC2 1等位基因可能与中国人 2型糖尿病视网膜并发症相关     

Androgen receptor gene CAG(n) trinucleotide repeats polymorphism in Chinese women with polycystic ovary syndrome

Trinucleotide repeats CAG(n) in androgen receptor gene is thought to be a potential site of genetic susceptibility to polycystic ovary syndrome (PCOS). However, previous studies of PCOS have shown variable association of CAG(n )polymorphism with PCOS. In order to evaluate CAG(n )polymorphism in Chinese women with PCOS, we have genotyped CAG(n) repeat numbers in female Chinese subjects (148 PCOS patients and 104 control subjects). The mean CAG(n) repeat lengths of PCOS patients and control subjects were similar (22.88 +/- 1.76 vs. 22.85 +/- 1.60; P = NS). No difference in the mean CAG(n) repeat lengths of hyperandrogenic and nonhyperandrogenic subgroups of PCOS patients was found (22.86 +/- 1.68 vs. 22.91 +/- 1.84; P = NS). Moreover, no difference was found in the term of mean CAG(n) repeat lengths in the nonhyperandrogenic subgroup and the control subjects (22.86 +/- 1.68 vs. 22.85 +/- 1.60; P = NS). However, mean CAG(n) repeat lengths were negatively correlated with serum total cholesterol and low-density lipoprotein-cholesterol concentration in PCOS patients (r = -0.182, P < 0.05 and r = -0.210, P < 0.05, respectively), but not with total testosterone, body mass index, waist and hip circumferences. The CAG(n) repeat length polymorphism may not be a major determinant of PCOS, but it may influence the lipid metabolism of PCOS patients.     

Association of tri-nucleotide (CAG and GGC) repeat polymorphism of androgen receptor gene in Taiwanese women with refractory or remission rheumatoid arthritis

We investigated the relationship between CAG and GGC repeat polymorphism of the androgen receptor (AR) gene and rheumatoid arthritis (RA) in female patients with different disease subtypes. This case-control study enrolled 215 women in three groups: RA patients refractory to standardized therapy (n = 51); RA patients at complete remission phase (n = 60); and healthy controls (n = 104). CAG and GGC repeat lengths were determined by automated fluorescence-based DNA fragment-sizing method. Demographic data, allele lengths, allele distribution, and zygosity status of CAG/GGC repeats were assessed for the three groups. Refractory RA patients tend to have a significantly younger onset age of RA and more elevated erythrocyte sedimentation rates than do remission RA patients. Mean and median values of CAG and GGC repeat lengths are similar in both RA and control patients. However, RA patients harboring any long CAG alleles with more than 23 repeats had an increased risk of a refractory course, whereas differences in risk were not observed between these patients and RA subtypes harboring any long GGC alleles with more than 16 repeats. In addition, the homozygous frequency of CAG but not GGC alleles was lower in refractory RA than in remission RA patients or in controls (p = 0.042). Neither CAG nor GGC repeat lengths had a significant relationship with rheumatoid factor reactivity. Our observations indicate that short CAG repeats of the AR gene with higher transactivation activity may have protective effects against refractory course of RA development and that homozygous frequency of CAG alleles may be involved in the disease remission subtype. In contrast, lack of association of GGC polymorphism and RA was also observed. Together, these data imply that CAG but not GGC alleles in the AR polymorphism may play an important role in modulating the disease pattern of RA among Taiwanese women.     

Correlation study between the polymorphism of repetitive sequence in gene CAG of androgen receptor and the occurrence and progression of prostate cancer

Objective:To explore the relation between the polymorphism of repetitive sequence in gene CAG of androgen receptor(AR)and the susceptibility and clinical stages as well as pathological grading of prostate cancer among Han population.Method:Sixty-eight cases with prostate cancer hospitalized in Urinary Surgery Department from Feb.2010 to Feb.2012 and 60 healthy cases were chosen as research subjects.Methods of PCR and direct sequencing were adopted to detect DNA sequence of AR gene and the length of repetitive sequence in CAG.Results:The lengths of repetitive sequence in CAG of patients with prostate cancer and healthy people were(22.3±4.6)and(23.0±4.9),respectively showing no statistical significance.Comparing length(repetitive sequence of CAG)22,those with that22 suffer a remarkably higher risk of prostate cancer(P0.05).The number of repetitive sequence in CAG of patients at clinical stage C-D was less than that of patients at stage B,and the number of repetitive sequence in CAG of patients with poorly differentiated prostate cancer was also less than that of patients with moderately and highly differentiated prostate cancer.But there was no statistical significance int the difference(P0.05);the proportion of patients with length22 at clinical stage C-D was much larger than that of patients at clinical stage B(P0.05),and as the aggravation of pathological grading,the proportion of patients with the length22 was also remarkably increased and there was significant difference between patients with highly differentiated prostate cancer and those with poorly differentiated prostate cancer(P0.05).Conclusions:There is correlation between the occurrence and development of prostate cancer in Han population and the polymorphism of repetitive sequence in gene CAG of androgen receptor.The less the number of repetitive sequence in CAG is,the higher the risk of prostate cancer will be and the more severe the clinical stage and pathological grading will be.     

Tandem CAG repeats of the androgen receptor gene and prostate cancer risk in black and white men

The most common malignancy in men worldwide is cancer of the prostate. Androgens play a direct role in normal and malignant growth of prostate cells via the androgen receptor (AR). This study analyzed the polymorphic CAG repeat sequence in exon 1 of the AR gene to determine if the number of repeats might be an indicator of prostate cancer risk or aggressive disease. DNA was extracted from blood samples of 20 black and 20 white men with well-documented prostate cancer and 40 healthy, controls (20 blacks and 20 whites). PCR amplifications was followed by gel electrophoresis and DNA sequencing. This region normally contains between 9 and 29 repeats. Patients and controls both had minor variations in the number of repeats, which ranged from 13 to 27 with 21 being the most frequent allele. Black controls and patients both had a mean of 20±3, repeats; in whites the mean was significantly lower in patients than controls (21±2 versus 23±2; p=0.004). Combined black and white patients also had a lower number than the combined group of controls (20±3 versus 22±3; p=0.02). Similarly, black and white patients with aggressive disease has a lower number than patients whose disease was more slowly progressive (19±2 versus 22±3; p=0.02). We conclude that the small differences in the number of CAG repeats in both black and white patients do not appear to be a strong indicator of risk or aggressive disease but that this size polymorphism may be one of many genetic and environmental risk factors involved in prostate cancer.     

Analysis of the Distribution of CAG Repeats and X-Chromosome Inactivation Status of HUMARA Gene in Healthy Female Subjects Using Improved Fluorescence-Based Assay

We investigated the polymorphic CAG-repeat distribution and the X-inactivation status of the human androgen receptor (HUMARA) gene in 58 female Japanese volunteers. Polymerase chain reaction amplification was performed using a fluorescent-dye-labeled primer under conditions specific for GC-rich targets, and fragments were analyzed. To estimate the length of these fragments, FAM-labeled (blue fluorescent) products were simultaneously compared with ROM-labeled size markers (red) that were created by sequencing various HUMARA fragments. The number of polymorphic CAG repeats of HUMARA in 116 alleles from 58 female subjects ranged from 15 to 28. Of the 58 volunteers, 51 (88.0%) were heterozygous. In 96% of the heterozygous female subjects, the allelic differences were no greater than 6 repeats. X-chromosome inactivation was calculated as the ratio of the area of the smaller peak to the sum of the areas of the smaller and larger peaks. The average ratio was 0.38 (range, 0.09-0.50). Preferential use of 1 allele, by more than 75% (ratio. <0.25). was observed in 5 volunteers (10.9%). The clonal nature of a patient with chronic myelogenous leukemia was easily identified. This method is sensitive enough to discriminate a difference of 1 triplet repeat.     

Combined profile of the tandem repeats CAG, TA and CA of the androgen and estrogen receptor genes in breast cancer

Purpose Case–control studies have reported inconsistent results concerning the association between polymorphisms in the androgen and estrogen receptor (ER) genes and breast cancer. While several studies investigated the association between the androgen receptor (AR) gene, CAG repeat and breast cancer, for the CA and TA repeats in the ER genes there are considerably fewer studies (one for CA and none for TA).Methods We have investigated the potential link between three tandem repeats (CAG, TA, and CA) in the AR, ERs α and β genes, respectively, and breast cancer. DNA was isolated from 153 invasive breast tumors and 318 controls, and the three tandem repeats were sized by polyacrylamide electrophoresis. Number of repeats in each allele and the total repeats of both alleles were taken as variables for classification into dichotomous groups using the median of each variable in the control group as cut-off point. Relationship between polymorphic tandem repeats and breast cancer was assessed by multivariate logistic regression models.Results Three variables combined, longer CAGsum (≥28), shorter TA (<23) and CA (<23) repeats could constitute a possible genetic profile associated with breast cancer.Conclusions Our results confirm previous reports regarding an association between longer CAG repeats and breast cancer. In addition to that, we found that the combination of long CAG, short TA and CA repeats are strongly associated with breast cancer.This paper was initially submitted with an additional co-author who withdrew the co-authorship during the review process. All listed authors have now signed that they agree with the content of the current paper.     

Insulin Receptor Gene in Hypertension

The first molecular genetic association with human essential hypertension (HT) involved the insulin receptor gene (INSR). This highly significant result in Caucasians was for an insertion/deletion polymorphism in intron 9. A polymorphism in exon 8 showed a weak association, but a microsatellite in intron 2 proved negative for HT, although has shown an association with plasma insulin in Japanese. A similar spectrum of genetic associations for variants spanning INSR has been noted for insulin-dependent diabetic patients with rapidly-progressing renal disease, a subgroup having a strong family history of essential HT. Association with HT has also been found for an INSR variant in Chinese. Insulin resistance secondary to an INSR ‘defect’, or other causes, would increase insulin, which has cardiovascular effects, and insulin can raise angiotensinogen. Also, insulin is co-secreted with amylin, which can increase renin secretion. In the spontaneously HT rat there is evidence for reduced down-regulation of INSR expression in response to NaCl-loading, consistent with a promoter effect. When combined with observations of insulin resistance in essential HT patients and their pre-HT offspring, the possibility of dys-regulation of INSR merits attention in disease etiology in a proportion of essential HT patients.