基于单细胞转录组测序数据分析PECAM1对胃肠道间质瘤细胞迁移和侵袭能力的影响

Analysis of the effect of PECAM1 on the migration and invasion of gastrointestinal stromal tumor cells based on single-cell transcriptome sequencing data

目的:探讨血小板和内皮细胞黏附分子1(platelet and endothelial cell adhesion molecule 1,PECAM1)表达对胃肠道间质瘤(gastrointestinal stromal tumor,GIST)细胞迁移和侵袭能力的影响。方法:基于GIST单细胞转录组测序数据,利用R语言筛选目标基因;细胞转染siRNA建立低表达实验组;伤口愈合实验、Transwell实验验证细胞迁移和侵袭能力的变化。结果:根据免疫细胞标志物PTPRC是否表达,将18个GIST细胞分群中的6个分群定义为非免疫细胞群;根据肿瘤干细胞标志物CD44、CD34和ABCB1的差异表达水平,将非免疫细胞群进一步划分成4个肿瘤干细胞亚群;对4个肿瘤干细胞亚群的差异表达基因作KEGG分析筛选出PECAM1基因;进一步基因集富集分析结果显示PECAM1与细胞迁移信号通路相关;细胞实验结果表明,相较于对照组,PECAM1表达降低后,细胞迁移和侵袭能力显著降低(P<0.05)。结论:PECAM1正向调控GIST细胞迁移和侵袭过程。

Objective:To investigate the effect of platelet and endothelial cell adhesion molecule 1(PECAM1)expression on migration and invasion ability in gastrointestinal stromal tumor(GIST)cells.Methods:The target gene was screened based on GIST single-cell transcriptome sequencing data by R language.Cells were transfected with siRNA to establish a low expression group.The effect of PECAM1 on cell migration and invasion was analyzed by wound healing assay and Transwell assay.Results:Six of eighteen GIST cell clusters were defined as non-immune cell populations based on the positive expression of PTPRC.According to the different expression levels of cancer stem cell markers CD44,CD34 and ABCB1,the non-immune cell populations were further divided into four cancer stem cell subpopulations.PECAM1 gene was screened by KEGG analysis with differential expression genes in four cancer stem cell subpopulations.Further gene set enrichment analysis showed that PECAM1 was associated with cell migration signaling pathway.Cell experiments showed that cell migration and invasion ability were significantly decreased after inhibiting expression level of PECAM1,compared to the negative control group(P<0.05).Conclusion:PECAM1 positively regulates cell migration and invasion in GIST cells.