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CRELD1基因过表达对心脏瓣膜相关基质蛋白的影响 被引量：1

Effects of overexpression of CRELD1 gene on heart valve-related matrix proteins
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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-TimePCR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。 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（P0.05）,while there was no significant difference in the expression of Tenascin-C mRNA and protein among groups（P0.05）.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.
作者 陈璇 郭颖 袁浪 张丽 黄敏 沈捷
机构地区 上海交通大学附属儿童医院上海市儿童医院心内科 上海交通大学医学院附属上海儿童医学中心心内科
出处 《上海交通大学学报（医学版）》 CAS CSCD 北大核心 2012年第3期247-251,共5页 Journal of Shanghai Jiao tong University:Medical Science
基金 国家自然科学基金(30772348) 浦东新区科技发展基金创新资金~~
关键词 CRELD1基因 过表达 TENASCIN-C AGGRECAN 房室间隔缺损 CRELD1 gene overexpression Tenascin-C Aggrecan atrioventricular septal defect
分类号 Q78 [生物学—分子生物学] R541.1 [医药卫生—心血管疾病]
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参考文献13

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共引文献71

同被引文献25

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上海交通大学学报（医学版）
2012年 第3期

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