基于基因芯片整合分析筛选及验证BeWo细胞合体化相关调控因子

Screening and validation of the regulatory factors for BeWo cell fusion based on gene microarray integration

摘要:

目的：合体滋养层细胞（syncytiotrophoblast，STB）是一类多核化上皮细胞位于人类胎盘最外层，由单核滋养细胞（cy－totrophoblast，CTB）分化、融合而来，肩负妊娠维持关键激素的分泌和母胎间营养交换的重要职责，也是母胎屏障重要组成部分之一。然而，目前关于胎盘滋养细胞的合体化过程的调控机制仍不明确。本研究即利用生物信息学的方法筛选并初步验证潜在调控绒毛膜癌BeWo细胞合体化调控的关键基因。方法：从GEO数据库检索并下载BeWo细胞合体化前后的基因芯片数据集，并利用生物信息学分析方法对所有相关数据集进行综合分析，筛选出差异表达基因并进行GO及KEGG聚类分析，进一步通过PPI蛋白互作网络寻找节点基因，最后利用qRT-PCR验证候选基因在合体化前后的差异表达。结果：从2组数据中共筛选出137个差异表达基因，其中25个差异基因为2组数据集共有，参与细胞迁移、细胞黏附、激素代谢、MAPK信号通路等。进一步借助蛋白质互作网络分析，得到3个候选基因并通过qRT-PCR验证，结果与信息分析结果一致。结论：本研究为滋养细胞合体化研究提供了潜在靶基因，也为解析其分子调控机制提供了线索。

Abstract:

Objective：To investigate the screening and preliminary validation of potential key genes for the regulation of BeWo cell fusion based on bioinformatics，since syncytiotrophoblasts（STBs） are a type of multinucleated epithelial cells located at the outermost layer of the placenta and are generated by the differentiation and fusion of cytotrophoblasts（CTBs）. STBs play an important role in the secretion of key hormones for pregnancy maintenance and nutrition exchange between mother and fetus during pregnancy. However，the regulatory mechanism of placenta trophoblast cell fusion remains unclear. Methods：The microarray datasets of BeWo cells before and after cell fusion were searched and downloaded from GEO database. A bioinformatics analysis was performed for a comprehensive analysis of all datasets to screen out the differentially expressed genes（DEGs），and then a cluster analysis was performed based on GO and KEGG. The PPI network was used to screen out node genes，and qRT-PCR was used to verify the differential expression of can－didate genes before and after cell fusion. Results：A total of 137 differentially expressed genes were screened out from two datasets，among which 25 were shared by the two datasets and were involved in cell migration，cell adhesion，hormone metabolism，and the MAPK signaling pathway. Three candidate genes were screened out by the PPI network and validated by qRT-PCR，and the results were consistent with the information analysis. Conclusion：This study provides some potential target genes for the research on trophoblast fusion and gives a clue to molecular regulation mechanism.