泸州医学院学报
瀘州醫學院學報
로주의학원학보
JOURNAL OF LUZHOU MEDICAL COLLEGE
2013年
4期
328-332
,共5页
张旭%江燕%张翠薇%荣智利%高岑%马跃荣
張旭%江燕%張翠薇%榮智利%高岑%馬躍榮
장욱%강연%장취미%영지리%고잠%마약영
E-cadherin%α-SMA%甲基化%上皮-间质转分化%肾小管上皮细胞
E-cadherin%α-SMA%甲基化%上皮-間質轉分化%腎小管上皮細胞
E-cadherin%α-SMA%갑기화%상피-간질전분화%신소관상피세포
E-cadherin%α-SMA%Methylation%Renal tubular epithelial cells%Epithelial-mesenchymal transition
目的:了解大鼠肾小管上皮细胞间质转分化与E-钙黏附蛋白(E-cadherin)和α-平滑肌肌动蛋白(α-SMA)基因启动子区甲基化状态之间的关系。方法:将大鼠肾小管上皮细胞分为对照组和TGF-β1处理组。对两组细胞采用免疫细胞化学方法检测E-cadherin和α-SMA蛋白表达情况,用ELISA法检测培养液中Ⅰ型胶原蛋白的含量。同时分别提取两组细胞中的基因组DNA,应用焦磷酸测序技术对E-cadherin和α-SMA基因启动子区甲基化水平进行检测。结果:TGF-β1处理组细胞中E-cadherin基因启动子区有2个CpG位点甲基化频率高于对照组,而α-SMA基因启动子区有2个CpG位点甲基化频率低于对照组。结论:E-cadherin和α-SMA基因启动子区甲基化状态的改变参与了肾小管上皮细胞间质转分化过程。
目的:瞭解大鼠腎小管上皮細胞間質轉分化與E-鈣黏附蛋白(E-cadherin)和α-平滑肌肌動蛋白(α-SMA)基因啟動子區甲基化狀態之間的關繫。方法:將大鼠腎小管上皮細胞分為對照組和TGF-β1處理組。對兩組細胞採用免疫細胞化學方法檢測E-cadherin和α-SMA蛋白錶達情況,用ELISA法檢測培養液中Ⅰ型膠原蛋白的含量。同時分彆提取兩組細胞中的基因組DNA,應用焦燐痠測序技術對E-cadherin和α-SMA基因啟動子區甲基化水平進行檢測。結果:TGF-β1處理組細胞中E-cadherin基因啟動子區有2箇CpG位點甲基化頻率高于對照組,而α-SMA基因啟動子區有2箇CpG位點甲基化頻率低于對照組。結論:E-cadherin和α-SMA基因啟動子區甲基化狀態的改變參與瞭腎小管上皮細胞間質轉分化過程。
목적:료해대서신소관상피세포간질전분화여E-개점부단백(E-cadherin)화α-평활기기동단백(α-SMA)기인계동자구갑기화상태지간적관계。방법:장대서신소관상피세포분위대조조화TGF-β1처리조。대량조세포채용면역세포화학방법검측E-cadherin화α-SMA단백표체정황,용ELISA법검측배양액중Ⅰ형효원단백적함량。동시분별제취량조세포중적기인조DNA,응용초린산측서기술대E-cadherin화α-SMA기인계동자구갑기화수평진행검측。결과:TGF-β1처리조세포중E-cadherin기인계동자구유2개CpG위점갑기화빈솔고우대조조,이α-SMA기인계동자구유2개CpG위점갑기화빈솔저우대조조。결론:E-cadherin화α-SMA기인계동자구갑기화상태적개변삼여료신소관상피세포간질전분화과정。
Objective:To investigate the relationship between renal tubular epithelial-mesenchymal transition and promoter methylation status of E-cadherin and α-SMA gene. Methods: Renal tubular epithelial cells of rat were divided into control group and TGF-β1 treated group. E-cadherin and α-SMA expression were detected by immunocytochemical staining; type Ⅰ collagen concentration in culture medium was tested by ELISA method;meanwhile genomic DNA was extracted, then the E-cadherin and α-SMA promoter methylation status was detected using pyrosequencing technique. Results: There was higher methylation level and lower methylation level in promoter region ofα-SMA gene in TGF-β1 group than in control group. Conclusion:Methylation changes in E-cadherin andα-SMA gene promoter region is involved in renal tubular epithelial-mesenchymal transition.