Lnc RNA TUC338影响人类宫颈癌细胞的蛋白组学分析

Proteomics Analysis of Lnc RNA TUC338 Affecting Human Cervical Cancer Cells

目的探究长链非编码RNA TUC338影响人类宫颈癌细胞的分子机制。方法将人类宫颈癌细胞Hela细胞分为实验组(TUC338组)与对照组(rLV组),分别感染rLV-hTUC338-ZsGreen-Puro慢病毒及rLV-ZsGreen-Puro慢病毒,RT-PCR检测两组细胞Lnc RNA TUC338表达量,并对两组细胞进行蛋白质谱分析。结果感染慢病毒后所有细胞均有绿色荧光显示,RT-PCR结果显示TUC338组Lnc RNA TUC338表达量较rLV组增加,两组均显示较多的蛋白条带;非标记定量法蛋白测序共检测到4323个蛋白。蛋白差异分析显示:与rLV组比较,在TUC338组中下调的蛋白有194个,上调的蛋白有225个。其中,重要的下调蛋白有:LAMB1、HNRNPH3、COX2等,上调蛋白有:EPPK1、ITPR1、AKR1C2等。最显著的下调GO为磷酸化修饰功能,上调GO为细胞骨架功能,最显著的下调KEGG通路为半乳糖代谢通路,上调KEGG通路为肌动蛋白细胞骨架调节通路。共检测到含WD40 repeat结构域(WD40 repeat)等20个蛋白结构域。rLV组和rLV-TUC338组中有显著差异的结构域包括Haem peroxidase,animal等。主要亚细胞成分及主要差异亚细胞成分均为核蛋白,分别占34.23%、24.22%。rLV组和rLVTUC338组中下调的互作网主要包括:prot1(P37268)-prot2(O95864)、prot1(Q6FGH9)-prot2(Q13363)等。上调的互作网主要包括:prot1(P60520)-prot2(A0A024RBQ5)、prot1(P60520)-prot2(Q06330)等。结论Lnc RNA TUC338影响了人类宫颈癌细胞的蛋白组学,是宫颈癌的抑癌基因,并通过糖代谢、磷酸化、细胞骨架这3个通路抑制肿瘤发展。

Objective To explore the molecular mechanism of long-chain non-coding RNA TUC338 affecting human cervical cancer cells.Methods Human cervical cancer Hela cells were divided into experimental group(TUC338 group)and control group(rLV group).rLV-hTUC338-ZsGreen-Puro lentivirus and rLV-ZsGreen-Puro lentivirus were transfected respectively.The expression of Lnc RNA TUC338 in the two groups was detected by RT-PCR,and the protein spectrum of the two groups was analyzed.Results All cells were fluorescently green after infection with lentivirus.The results of RT-PCR showed that the expression of Lnc RNA TUC338 in TUC338 group was higher than that in rLV group,and both groups showed more protein bands.A total of 4323 proteins were detected by non-labeled quantitative protein sequencing.Protein difference analysis showed that compared with the rLV group,194 proteins were down-regulated and 225 proteins were up-regulated in the TUC338 group.Among them,the important down-regulated proteins were LAMB1,HNRNPH3,COX2 and up-regulated proteins were EPPK1,ITPR1,AKR1C2 and so on.The most significant down-regulation of GO was phosphorylation modification function,and up-regulation of GO was cytoskeleton function.The most significant down-regulation of KEGG pathway was galactose metabolism pathway,and up-regulation of KEGG pathway was actin cytoskeleton regulation pathway.A total of 20 protein domains including WD40 repeat domain were detected.In r LV group and rLV-TUC338 group,there were significant differences in domains including Haem peroxidase,animals,etc.The main subcellular components and the main differential subcellular components were nuclear proteins,accounting for 34.23% and 24.22%,respectively.The down-regulated interaction network in rLV group and rLV-TUC338 group mainly included:prot1(P37268)-prot2(O95864),prot1(Q6FGH9)-prot2(Q13363),etc.The up-regulated interaction network mainly included:prot1(P60520)-prot2(A0A024RBQ5),prot1(P60520)-prot2(Q06330),etc.Conclusion Lnc RNA TUC338 affects the proteomics of human cervical cancer cells,is a tumor suppressor gene of cervical cancer,and inhibits tumor development through glucose metabolism,phosphorylation and cytoskeleton.