FABP3对P19细胞增殖的影响及其生物信息学分析

Effect of FABP3 on proliferation of P19 cells and bioinformatics analysis of FABP3

目的探讨脂肪酸结合蛋白(FABP)3基因过表达对P19细胞分化、增殖的影响及FABP3基因的生物信息学特征。方法构建FABP3过表达载体及相应的空载体,转染P19细胞,经0.9%二甲基亚砜(DMSO)诱导分化为心肌细胞,定量PCR检测肌钙蛋白T(cTnT)和转录调控因子GATA4基因的表达,MTT法检测P19细胞增殖,应用在线数据库软件分析FABP3的核苷酸基本特征、蛋白质理化性质、亚细胞定位等。结果与空载体相比,FABP3基因过表达能抑制cTnT、GATA4 mRNA表达及P19细胞增殖(P<0.05或P<0.01)。生物信息学分析显示,FABP3基因mRNA全长1097 bp,开放阅读框长402 bp,编码133个氨基酸,相对分子质量为14 858 Da,蛋白主要位于胞浆。结论 FABP3基因过表达显著抑制心肌细胞分化、增殖,生物信息学分析为进一步研究该基因奠定了基础。

Objective To investigate the effect of overexpression of fat acid binding protein（FABP）3 on the differentiation and proliferation of P19 cells and the bioinformatic characteristics of FABP3 gene.Methods The overexpressing vector of FABP3 and its corresponding empty vector were constructed and transfected into P19 cells.Then the P19 cells overexpressed FABP3 were induced by 0.9% DMSO.The expression levels of cardiac troponin T（cTnT） and zinc finger-containing transcription factor GATA4 gene were detected by RT-PCR and the proliferation of P19 cells was assayed by MTT method.The basic characteristic of nucleotide,physico-chemical property of protein and subcellular localization of FABP3 were analyzed by online database software.Results Compared with empty vector,overexpressed FABP3 gene could inhibit the mRNA expressions of cTnT and GATA4 and the proliferation of P19 cells（P〈0.05 or P〈0.01）.Bioinformatics analysis showed that the total length of FABP3 mRNA was 1097 bp and the open reading frame was 402 bp with encoding of 133 amino acids and relative molecular mass of 14 858 Da,and FABP3 mainly localized in the cytoplasm.Conclusion Overexpressed FABP3 gene significantly inhibits the differentiation and proliferation of P19 cells and the bioinformatics analysis may provide the physical and chemical informations for further study.