趋化因子CCL5在肾透明细胞癌发生发展中的作用

Role of the chemokine family member CCL5 in the development of clear cell renal cell carcinoma

目的 探讨趋化因子CCL5 (C-C chemokine ligand 5)在肾透明细胞癌(clear cell renal cell carcinoma,ccRCC)发生发展中的作用。方法 使用生物信息挖掘技术对GSE36895、GSE46699、GSE53757基因表达谱芯片数据集进行分析后取交集得到差异表达基因(differential expressed genes,DEGs),使用STRING数据库及Cytoscape的cytoHubba应用程序得到蛋白质互作网络(protein-protein interaction networks,PPI)及关键基因并进行排序,最后将所得关键基因在癌症基因组图谱(the cancer genome atlas,TCGA)及基因型-组织表达(genotype-tissue expression,GTEx)数据库中进行验证。随后使用细胞及动物实验验证CCL5对透明细胞癌细胞增殖的影响。结果 通过对GEO (Gene Expression Omnibus)数据库ccRCC的三个数据集使用R软件等工具进行分析,共得到934个DEGs,其中上调504个,下调430个。构建DEGs的PPI互作网络并通过cytoHubba筛选出最重要的10个基因并与TCGA及GTEx数据库数据进行对比,研究发现了趋化因子家族相关基因在ccRCC发生发展中起重要作用。与空白对照组比较,EdU检测提示CCL5沉默组786-o细胞的增殖显著减少(P<0.05),CCK8检测提示CCL5沉默组786-o细胞的增殖显著亦减少(P<0.001)。通过小鼠异种移植肿瘤模型实验发现,相较空白对照组,在第50天,CCL5沉默组的肿瘤体积明显低于对照组(P<0.001)。结论 趋化因子CCL5在ccRCC发生发展中起到了重要作用。

Objective To investigate the role of the chemokine family member CCL5 in the development of clear cell renal cell carcinoma(ccRCC).Methods The microarray data sets of GSE36895,GSE46699,and GSE53757 gene expression profiles were analyzed by using bioinformation mining technology,and the cross sets were obtained to find differential expressed genes(DEG).Using the STRING database and cyto Hubba of Cytoscape gets PPI(protein-protein interaction) network and hub genes.The hub genes are verified in TCGA(the cancer genome atlas) and GTEx(genotype-tissue expression) database.Cell and animal experiments were used to verify the role of CCL5 in the proliferation of ccRCC.Results Three data sets of ccRCC in GEO(Gene Expression Omnibus) database were analyzed by using R software and other tools,a total of 934 DEGs(differential expressed genes) were found,of which504 were up-regulated and 430 down-regulated.The PPI interaction network of DEGs was constructed and the most important 10 genes were screened out by cytohubba and compared with TCGA and GTEX database data.We found that chemokine family genes play an important role in the occurrence and development of prostate cancer.The EdU tests results showed that the proliferation of 786-o cells was significantly reduced in the CCL5 silencing group compared with the blank control group(P<0.05).CCK8 tests indicated that the proliferation of 786-o cells in the CCL5 silent group was significantly decreased compared with the blank control group(P<0.001).Mouse xenografted tumor models showed that the tumor volume in the CCL5 silencing group was significantly lower than that in the control group on day 50(P<0.001).Conclusion Through deep biological information mining,chemokine family member CCL5 plays an important role in the development of ccRCC.