内质网自噬受体1对胃癌细胞生物学功能的影响

Effect of FAM134B on biological functions of gastric cancer cells

目的内质网自噬受体1(FAM 134 B)在胃癌中的具体作用机制尚不明确。文中旨在研究FAM 134 B在胃癌细胞中表达上调对胃癌细胞增殖、迁移、侵袭和凋亡的影响,并分析FAM 134 B相关的信号通路及相互作用蛋白对FAM 134 B生物学功能。方法构建FAM 134 B过表达的小鼠胃癌MFC细胞系模型,将含有pcDNA3.1-FAM134B的MFC细胞系(FAM 134 B组)滴加到6孔板中继续培养;另于MFC细胞系中转染pcDNA3.1-vector质粒(对照组),1 d后行蛋白印迹实验评估质粒转染情况。通过细胞划痕实验检测FAM 134 B表达上调对胃癌细胞迁移的影响,通过CCK8法检测FAM 134 B表达上调对胃癌细胞增殖的影响,通过Transwell法检测FAM 134 B表达上调对胃癌细胞侵袭的影响,通过Annexin P/PI双染色法检测FAN134B表达上调对胃癌细胞凋亡的影响。最后利用基因富集分析和构建相互作用蛋白网络图对FAM 134 B相关生物学功能进行探索。结果FAM 134 B组细胞24 h后的愈合率[(48.333±7.095)%]较对照组[(72.667±4.509)%]明显降低(P=0.018)。FAM 134 B组A 450nm波长的密度值(0.643±0801)较对照组(1.071±0.783)明显降低(P=0.008)。FAM 134 B组细胞计数[(34.750±3.500)个]较对照组[(57.000±4.690)个]明显降低(P=0.001)。FAM 134 B组细胞凋亡数较对照组明显升高(P=0.007)。富集涉及178个基因集共56514个基因,随FAM 134 B表达水平上升而上调包括102个基因集,且FAM 134 B主要参与了32条信号通路(P<0.05,FDR<0.05)。结论FAM 134 B表达上调可以抑制胃癌细胞的增殖、迁移、侵袭,并促进胃癌细胞的凋亡,且FAM 134 B相关信号通路及相互作用蛋白密切参与肿瘤的进展,FAM 134 B可能成为胃癌诊疗过程中重要的靶点。

Objective The specific mechanism of FAM134B in gastric cancer is still unclear.This study aims to study the effect of the up-regulated expression of FAM134B on the proliferation,migration,invasion and apoptosis of gastric cancer cells,and to analyse the signal pathways and interacting proteins related to FAM134B on the biological functions of FAM134B.Methods A mouse gastric carcinoma cell line MFC with over-expression of FAM 134 B was established.MFC cell line with pcDNA3.1-FAM124B was cultured in 6-well plates(FAM134B group).In MFC cell line,pcDNA3.1-vector was transfected(control group),which was detected by Western blot after one day.The effect of up-regulation of FAM 134 B expression on migration,proliferation,invasion and apoptosis of gastric cancer cells,was respectively detected by cell scratch assay,CCK8 assay,Transwell assay and Annexin P/PI double staining.Ultimately,the gene enrichment analysis(GSEA)and interaction protein network mapping were used to further explore the biological functions of FAM134B.Results After 24 hours in vitro experiment,the healing rate,the density of wavelength and the cells of FAM134B group were lower.Meanwhile,the apoptosis of FAM134B was higher.GSEA has 178 gene-sets and 56514 genes.With the up-regulated expression of FAM134B including 102 genes,FAM 134 B was mainly involved in 32 signaling pathways(P<0.05,FDR<0.05).Conclusion The up-regulated expression of FAM 134 B can inhibit the proliferation,migration and invasion of gastric cancer cells,and promote the apoptosis of gastric cancer cells.Moreover,the FAM134B-related signaling pathway and interacting proteins are closely involved in tumor progression.FAM 134 B may become an important target in the diagnosis and treatment of gastric cancer.