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|本期目录/Table of Contents|

下调GPM6B基因抑制平滑肌表达特异性标记蛋白

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2018年第5期
页码:
508-511,516
栏目:
基础研究
出版日期:
2018-06-25

文章信息/Info

Title:
Knockdown of GPM6B represses expression of smooth muscle cell specific genes
作者:
张晓萌1; 王 姗2; 常 盼3; 赵会寿2; 谢华宁4; 闫 凤2; 夏云龙2; 张静隆2; 张 玲2; 陶 凌2; 王海昌1
（空军军医大学:1.唐都医院心血管内科，陕西 西安 710038；2.西京医院心血管内科，陕西 西安 710032；3.西安医学院第二附属医院中心实验室，陕西 西安 710038；4.陕西省中医医院心病科，陕西 西安 710000）
Author(s):
ZHANG Xiao-meng1;  WANG Shan2;  CHANG Pan3;  ZHAO Hui-shou2;  XIE Hua-ning4;  YAN Feng2;  XIA Yun-long2;  ZHANG Jing-long2;  ZHANG Ling2;  TAO Ling2;  WANG Hai-chang1
(1.Department of Cardiology, Tangdu Hospital, Air Force Military Medical University, Xi’an 710038, Shaanxi, China; 2.Department of Cardiology, Xijing Hospital, Air Force Military Medical University, Xi’an 710032, Shaanxi, China; 3.Center Laboratory, Second Affiliated Hospital, Xi’an Medical University, Xi’an 710038, Shaanxi, China; 4.Department of Heart Disease, Shaanxi Traditional Medicine Hospital, Xi’an 710000, Shaanxi, China)
关键词:
GPM6B基因; 间充质细胞; 平滑肌; 分化; 慢病毒
Keywords:
GPM6B;  mesenchymal cells;  smooth muscle cells;  differentiation; lentivirus
分类号:
R542.2
DOI:
-
文献标识码:
A
摘要:
目的 研究糖蛋白M6B（Glycoprotein M6B,GPM6B）基因对间充质细胞C3H10T1/2向平滑肌细胞分化的影响。方法 将慢病毒感染后的sh-GPM6B细胞作为处理组，慢病毒感染后的sh-Scramble细胞作为对照组进行后续实验。用sh-GPM6B质粒和工具质粒在293T细胞系包装慢病毒。用病毒液感染C3H10T1/2小鼠间充质细胞系制备sh-GPM6B稳转细胞系。荧光显微镜观察病毒转染效率，Western法检测GPM6B基因的干涉效率。慢病毒shRNA干涉抑制GPM6B基因表达后，通过TGF-β1诱导对照组和sh-GPM6B细胞向平滑肌分化。Western blot法检测平滑肌特异标记蛋白SM-MHC和α-SMA表达水平。q-PCR法检测SM-MHC和α-SMA的mRNA转录水平。结果 慢病毒感染后sh-GPM6B干涉细胞GPM6B基因蛋白表达降低至对照组（40~70）%水平。Sh-GPM6B处理组细胞与sh-Scramble对照组细胞相比，SM-MHC蛋白表达水平显著下调（P<0.05）；SM-MHC和α-SMA mRNA转录水平明显降低（P<0.01）。结论 下调GPM6B基因可抑制TGF-β1诱导的间充质细胞向平滑肌细胞分化。
Abstract:
AIM To determine the effects of the Glycoprotein M6B (GPM6B) gene on mesenchymal cells differentiating into smooth muscle cells. METHODS Infected sh-GPM6B cells were used as treatment group cells, and sh-Scramle cells were used as a control group. sh-GPM6B plasmids and tool plasmids were used to pack lentivirus in the 293T cell line. The virus was used to infect C3H10T1/2 cells into a sh-GPM6B stable transferring cell line. A fluorescent microscope was used to detect transfer efficiency and Western blot was used to detect interference efficiency. After incubation with TGF-β1 for 3 days in control cells and sh-GPM6B cells, the expression and transcription levels of smooth muscle cell specific marker proteins (SM-MHC and α-SMA) were detected by Western blot and q-PCR. RESULTS Compared with the sh-Scramble control group cells, expression levels of GPM6B in the sh-GPM6B treatment group cells infected with lenti-sh-GPM6B virus decreased to 40-70%. Compared with the sh-Scramble control group cells, the expression levels of SM-MHC and α-SMA proteins in the sh-GPM6B treatment group cells was significantly decreased (P<0.01). The transcription levels of SM-MHC and α-SMA mRNA were significantly decreased (P<0.01). CONCLUSION Knockdown of GPM6B represses mesenchymal cells differentiating into smooth muscle cells induced by TGF-β1.

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备注/Memo

备注/Memo:
收稿日期：2017-11-26.基金项目：国家自然科学基金项目资助（81670204，81730011，81470478，81600683） 通讯作者：王海昌，主任医师，主要从事冠心病的诊治和基础研究 Email：wanghcz@fmmu.edu.cn 共同通讯作者：陶凌，主任医师，主要从事糖尿病心肌损伤研究 Email：lingtao@fmmu.edu.cn 作者简介：张晓萌，硕士生 Email：fmmu_zxm@qq.com
更新日期/Last Update: 1900-01-01