PRKG1基因非编码区多态与蒙古族原发性高血压的易感性研究

目的:高血压是由遗传和环境因素共同作用而引起的复杂性疾病,本研究预探讨PRKG1基因非编码区多态在蒙古族原发性高血压中的作用。方法:采集内蒙古自治区锡林郭勒盟蒙古族标本429例,其中高血压212例,正常血压者217例。对PRKG1基因的启动子和3’非翻译区进行测序后对突变位点进行基因分型。结果:PRKG1基因测序共发现3个突变位点,分别是PRKG1-1,PRKG1-2和PRKG1-3。在蒙古族人群中,PRKG1基因的PRKG1-2和PRKG1-3位点各检测到A等位基因和AA基因型,未检测到另一等位基因和和其对应的基因型,PRKG1-1位点的等位基因频率和基因型频率在高血压组和正常对照组间差异无统计学意义。结论:PRKG1基因启动子和3’非翻译区多态与蒙古族原发性高血压无明显相关性。

PRKG1在膀胱癌组织中的表达分析和临床意义研究

目的分析cGMP依赖性蛋白激酶1(PRKG1)在膀胱癌中的表达水平及其临床意义。方法获取TCGA数据库中膀胱癌和癌旁正常组织中PRKG1 mRNA表达信息,分析PRKG1在膀胱癌和癌旁正常组织中的表达水平。收集本院膀胱癌石蜡标本及膀胱癌患者的临床资料,采用免疫组化方法检测PRKG1蛋白表达水平,分析其表达和膀胱癌分期、分级等临床资料的相关性。同时在体外通过腺病毒感染细胞,获取稳定高表达PRKG1细胞后,评估PRKG1对膀胱癌细胞增殖和凋亡的调控。结果PRKG1在膀胱癌组织中表达水平下降,且低表达PRKG1和高临床分期、肌层浸润相关。体外实验结果表明,过表达PRKG1可诱导膀胱癌细胞凋亡,抑制细胞增殖。结论膀胱癌组织中PRKG1表达下降和高临床分期、分级相关,体外过表达PRKG1可调控细胞增殖、凋亡,PRKG1可能为膀胱癌潜在的生物标志物和生物治疗靶点。

PRKG1 mutation identified by whole-exome sequencing:a potential genetic etiology for He-Zhao deficiency

Objective:He-Zhao deficiency was originally described as a severe type of nonsyndromic hypodontia,and the causative gene locus was mapped to chromosome 10q11.2.The aim of this study was to identify potential genetic mutations that could cause He-Zhao deficiency.Methods:Patients with He-Zhao deficiency and their unaffected relatives of the large pedigree were investigated.The whole-exome sequencing using next-generation sequencing was employed to identify genetic variants.The data generated from the whole-exome sequencing using the Illumina Novaseq 6000 system were further analyzed by Burrows-Wheeler Aligner software,Sequence Alignment/Map tools and ANNOVAR tool.In vitro luciferase assay was used to investigate the effect of the detected mutation on gene expression.R environment was used to conduct t-tests.The study protocol was approved by the Research Ethics Committee of Bio-X Institutes,Shanghai Jiao Tong University(M2011004).Results:The exomes of five patients with He-Zhao deficiency and two of their unaffected relatives identified a mutation in PRKG1αas the molecular etiology of the disease.The variant c.-144 C>A of PRKG1 isoform 1 cosegregated with permanent tooth agenesis in 93 family members who were older than 12,at which time the primary teeth should have been replaced with permanent teeth.Functional studies suggested that the mutant allele promotes gene transcription by increasing its promoter activity.Conclusion:c.-144 C>A variant of PRKG1αinvolving odontoclast-associated root resorption is responsible for He-Zhao deficiency,unlike other forms of hypodontia,which typically involve odontoblast dysfunction.

抑制miR181b通过上调PKG-1减轻大鼠心肌细胞肥大

目的:通过检测miR181b在心肌肥厚患者外周血中的表达,研究miR181b与PRKG-1在心肌肥大中的作用机制,为心肌肥厚临床诊断和治疗提供新的靶点。方法:采用实时定量PCR方法检测50例临床诊断为心肌肥厚患者及25例正常健康人的外周血miR181b的水平,分析其表达水平与临床病理特征的关系。结果:miR181b在心肌肥厚患者外周血中表达(3.35±0.22)升高,差异有统计学意义(P<0.05);体外大鼠原代心肌细胞在镜下呈多角形,采用α-横纹肌肌动蛋白抗体和DAPI进行免疫荧光染色,镜下90%以上细胞发出绿色荧光,表明原代培养心肌细胞纯度较高;采用去氧肾上腺素处理心肌细胞72 h后,心肌细胞总蛋白表达显著升高(P<0.05);流式细胞术结果显示PE组心肌细胞大小明显增加(P<0.05);实时定量PCR检测结果表明,PE组中心肌肥大相关基因β-MHC、α-SA、ANP显著升高(P<0.05),证明体外构建心肌肥大模型成功。结论:miR181b在心肌肥厚患者外周血中表达显著上调;体外成功培养乳鼠原代心肌细胞,通过PE处理法成功构建体外心肌肥大模型;在肥大心肌细胞中,miR181b与prkg-1 m RNA及蛋白的表达有显著相关性;miR181b可能通过调控prkg-1 m RNA及蛋白水平的表达参与心肌肥大,有望成为心肌肥厚临床诊断和治疗的新的靶点。