型跨膜蛋白α亚型缺失突变体对人牙周膜成纤维细胞周期的影响

目的：研究内质网信号通路Ⅰ型跨膜蛋白α亚型（IRE1α）缺失突变体对人牙周膜成纤维细胞（hPDLFs）细胞周期的影响。方法在成功构建人IRE1α基因全长重组质粒的基础上，采用重叠聚合酶链反应构建其两个主要功能域Kinase和Rnase的缺失突变体（pD-Kinase、pD-Rnase）；然后分别染入hPDLFs细胞，Westernblot检测重组基因表达情况，流式细胞仪（FCM）检测转染后hPDLFs细胞的细胞周期变化。结果酶切及测序结果证实构建的IRE1α缺失突变体重组质粒构建成功；Westernblot分析结果显示，3种重组基因均能正确表达；FCM结果分析显示：与衣霉素（TM）组相比，IR...

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Published in华西口腔医学杂志 Vol. 32; no. 3; pp. 221 - 224
Main Author 李苹苹 罗俊 彭志庆 初颜兵 王燕
Format Journal Article
LanguageChinese
Published 重庆医科大学附属口腔医院牙体牙髓科,重庆,400015%重庆医科大学附属永川医院口腔科,重庆,402160%第三军医大学附属西南医院口腔科,重庆,400038%重庆医科大学附属口腔医院预防与儿童牙病科,重庆,400015 2014
Subjects
型跨膜蛋白α亚型
牙周膜成纤维细胞
细胞周期
缺失体
细胞周期
牙周膜成纤维细胞
缺失体
type Ⅰ transmembrane protein
deletion
cell cycle
型跨膜蛋白α亚型
periodontal ligament fibroblasts
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ISSN1000-1182
DOI10.7518/hxkq.2014.03.003

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Summary:目的：研究内质网信号通路Ⅰ型跨膜蛋白α亚型（IRE1α）缺失突变体对人牙周膜成纤维细胞（hPDLFs）细胞周期的影响。方法在成功构建人IRE1α基因全长重组质粒的基础上，采用重叠聚合酶链反应构建其两个主要功能域Kinase和Rnase的缺失突变体（pD-Kinase、pD-Rnase）；然后分别染入hPDLFs细胞，Westernblot检测重组基因表达情况，流式细胞仪（FCM）检测转染后hPDLFs细胞的细胞周期变化。结果酶切及测序结果证实构建的IRE1α缺失突变体重组质粒构建成功；Westernblot分析结果显示，3种重组基因均能正确表达；FCM结果分析显示：与衣霉素（TM）组相比，IRE1α实验组hPDLFs细胞S期比例增加而G1期减少（P〈0.05）；D-Rnase突变体组hPDLFs细胞S期比例减少而G1期增加（P〈0.05）；D-Kinase突变体组对hPDLFS细胞的增殖和各周期分布影响则无显著差异（P〉0.05）。结论在内质网应激状态下，IRE1α可促进hPDLFs细胞从G1期进入S期，D-Rnase突变体导致hPDLFs细胞生长阻滞于G1期，而D-Kinase则对hPDLFS细胞周期分布无明显影响。
Bibliography:51-1169/R
Li Pingping, Luo Jun, Peng Zhiqing, Chu Yanbing, Wang Yan （1. Dept. of Endodontics, The Affiliated Hospital of Sto- matology, Chongqing Medical University, Chongqing 400015, China; 2. Dept. of Stomatology, The Yongchuan Affiliated Hospital, Chongqing Medical University, Chongqing 402160, China; 3. Dept. of Stomatology, Southwest Hospital, The Third Military Medical University, Chongqing 400038, China; 4. Dept. of Preventive and Pediatric Dentistry, The Affiliated Hos- pital of Stomatology, Chongqing Medical University, Chongqing 400015, China）
type Ⅰ transmembrane protein;deletion;periodontal ligament fibroblasts;cell cycle
Objective To determine the effect of type Ⅰ transmembrane protein （IRE1α） deletions on the cell cycle of human periodontal ligament fibroblasts （hPDLFs） cells. Methods Based on the IRE1α deletions, a full-length model was successfully constructed. Moreover, overlapping polymerase chain reaction mutagenesis facilitated the establishment of two deletion mutants of IRE1α （pD-Kinase, pD
ISSN:1000-1182
DOI:10.7518/hxkq.2014.03.003