上海交通大学学报（医学版）

›› 2012, Vol. 32 ›› Issue (3): 247-.doi: 10.3969/j.issn.1674-8115.2012.03.003

• 论著（基础研究） • 上一篇    下一篇

CRELD1基因过表达对心脏瓣膜相关基质蛋白的影响

陈 璇1, 郭 颖2, 袁 浪1, 张 丽1, 黄 敏1, 沈 捷1   

  1. 1.上海交通大学附属儿童医院 上海市儿童医院心内科, 上海 200040； 2.上海交通大学 医学院附属上海儿童医学中心心内科, 上海 200127
  • 出版日期:2012-03-28 发布日期:2012-03-28
  • 通讯作者: 沈 捷, 电子信箱: shenxiaomin@yahoo.com。
  • 作者简介:陈 璇（1986—）, 女, 硕士生；电子信箱: chenxuan010101@163.com。
  • 基金资助:

    国家自然科学基金（30772348）；浦东新区科技发展基金创新资金

Effects of overexpression of CRELD1 gene on heart valve-related matrix proteins

CHEN Xuan1, GUO Ying2, YUAN Lang1, ZHANG Li1, HUANG Min1, SHEN Jie1   

  1. 1. Department of Cardiology, Shanghai Children's Hospital, Children's Hospital, Shanghai Jiaotong University, Shanghai 200040, China；2.Department of Cardiology, Shanghai Children's Medical Center, Shanghai Jiaotong University School of Medicine, Shanghai 200127, China
  • Online:2012-03-28 Published:2012-03-28
  • Supported by:

    National Natural Science Foundation of China, 30772348；Science and Technology Foundation of Pudong New District

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摘要：

目的 探讨CRELD1基因过表达对心脏瓣膜相关基质蛋白的影响。方法 采用PCR技术获得CRELD1基因，利用质粒pcDNA3.1(-)构建目的基因的真核表达载体，酶切及DNA测序鉴定。利用脂质体Lipofectamine 2000介导重组质粒转染人胚肺成纤维细胞株（HFL-I）（重组质粒组），以空载体转染细胞和未转染细胞分别作为转染对照组和空白对照组。采用Real-Time PCR技术和Western blotting方法检测细胞中心脏瓣膜基质蛋白Tenascin-C和Aggrecan 的mRNA和蛋白相对表达量。结果 重组质粒测序鉴定结果与GenBank数据库比对序列一致。重组质粒组HFL-I中Aggrecan的mRNA和蛋白相对表达量均显著低于转染对照组和空白对照组（P<0.05）；各组HFL-I中Tenascin-C的mRNA和蛋白相对表达量比较，差异均无统计学意义（P>0.05）。结论 CRELD1基因过表达时可以下调人胚肺成纤维细胞中心脏瓣膜基质蛋白Aggrecan的表达量，为阐明CRELD1基因在房室间隔缺损中的作用提供了一定的理论基础。

关键词: CRELD1基因, 过表达, Tenascin-C, Aggrecan, 房室间隔缺损

Abstract:

Objective To investigate the effects of overexpression of CRELD1 gene on heart valve-related matrix proteins. Methods CRELD1 gene was obtained by PCR. Target gene eukaryote expression vectors were constructed by pcDNA3.1 (-) vector plasmid, and were identified by restriction enzyme digestion and DNA sequence analysis. Recombinant plasmid was transfected into human embryonic lung fibroblasts （HFL-I）with lipofectamine 2000 (recombinant plasmid group), and HFL-I cells transfected with empty vectors and those without transfection were served as transfection control group and blank control group respectively. Real-Time PCR and Western blotting were employed to detect the relative expression of mRNA and protein of heart valve-related matrix proteins Tenascin-C and Aggrecan in each group. Results DNA sequence of recombinant plasmid agreed very well with that in GenBank according to DNA sequence analysis. The relative expression of Aggrecan mRNA and protein in recombinant plasmid group was significantly lower than that in transfection control group and blank control group (P＜0.05), while there was no significant difference in the expression of Tenascin-C mRNA and protein among groups (P>0.05). Conclusion CRELD1 gene overexpression can decrease the heart valve-related matrix protein Aggrecan in human embryonic lung fibroblasts, which serves as a theoretical framework to demonstrate the roles of CRELD1 gene on atrioventricular septal defect.

Key words: CRELD1 gene, overexpression, Tenascin-C, Aggrecan, atrioventricular septal defect