Xuan Xiuyun, Wang Yuqi, Sun Yanhong, Huang Changzheng
Department of Dermatology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430022, People's Republic of China.
Department of Radiology, Municipal Hospital Affiliated to Xuzhou Medical University, Xuzhou, Jiangsu, 221002, People's Republic of China.
Int J Gen Med. 2021 Nov 19;14:8495-8508. doi: 10.2147/IJGM.S336295. eCollection 2021.
Melanoma is one of the most malignant and aggressive skin tumors, and its incidence is increasing worldwide. However, few studies have investigated the process of tumorigenesis from normal skin to melanoma.
Several bioinformatics analyses, including GEO databases, Oncomine database, TCGA database, STRING, MCODE and cytoHubba plug-ins, GEPIA, TIMER and TRRUST and DGIdb, were performed to disclose the hub genes and immunology implicated in primary melanoma tumorigenesis. Finally, immunohistochemistry (IHC) and quantitative real-time PCR (qPCR) were used to validate the results of bioinformatics analysis in vitro.
A total of 295 overlapping DEGs (ODEGs) (157 upregulated and 138 downregulated) and 9 hub genes were identified between primary melanoma and normal skin tissues. Functional analysis of these 9 hub genes indicated that the genes were primarily enriched in cell chemotaxis, the chemokine-mediated signaling pathway, the extracellular region, the extracellular space, chemokine activity and CXCR3 chemokine receptor binding. KEGG pathway enrichment showed that these genes were primarily involved in the chemokine signaling pathway, cytokine-cytokine receptor interaction, the toll-like receptor signaling pathway, the cytosolic DNA-sensing pathway and the TNF signaling pathway. Upregulated CCL5, CCL4, CXCL9 and CXCL10 demonstrated good overall survival (OS), and most of them have a higher expression in stage 0 and 1 of melanoma. Moreover, immune infiltration analysis showed that the above hub genes showed a strong positive correlation between their expression and infiltration of the six immune cell subsets. Transcription factor regulation network suggested that RELA and NFKB1 are the transcription factors of CCL4, CCL5, CXCL10 and CXCL2, while IRF7, IRF3 and IRF1 are the transcription factors of CCL5 and CXCL10. Drug-gene interaction analysis identified 46 drug-gene interactions. In vitro data demonstrated that the level of CCL4, CCL5, CXCL9 and CXCL10 is higher in melanoma than that in normal skin tissues, either at tissue or cell lines level.
In summary, we identified 4 key chemokine members related to tumorigenesis and progression in primary melanoma, and these results may help to elucidate melanoma tumorigenesis and facilitate its treatment.
黑色素瘤是最具恶性和侵袭性的皮肤肿瘤之一，其在全球的发病率正在上升。然而，很少有研究调查从正常皮肤到黑色素瘤的肿瘤发生过程。
进行了多项生物信息学分析，包括GEO数据库、Oncomine数据库、TCGA数据库、STRING、MCODE和cytoHubba插件、GEPIA、TIMER以及TRRUST和DGIdb，以揭示原发性黑色素瘤肿瘤发生中涉及的关键基因和免疫学特征。最后，采用免疫组织化学（IHC）和定量实时聚合酶链反应（qPCR）在体外验证生物信息学分析结果。
在原发性黑色素瘤和正常皮肤组织之间共鉴定出295个重叠差异表达基因（ODEG）（157个上调和138个下调）以及9个关键基因。对这9个关键基因的功能分析表明，这些基因主要富集于细胞趋化性、趋化因子介导的信号通路、细胞外区域、细胞外空间、趋化因子活性以及CXCR3趋化因子受体结合。KEGG通路富集显示，这些基因主要参与趋化因子信号通路、细胞因子-细胞因子受体相互作用、Toll样受体信号通路、胞质DNA传感通路和TNF信号通路。上调的CCL5、CCL4、CXCL9和CXCL10显示出良好的总生存期（OS），并且它们中的大多数在黑色素瘤的0期和1期表达较高。此外，免疫浸润分析表明，上述关键基因的表达与其在六个免疫细胞亚群中的浸润之间呈强烈正相关。转录因子调控网络表明，RELA和NFKB1是CCL4、CCL5、CXCL10和CXCL2的转录因子，而IRF7、IRF3和IRF1是CCL5和CXCL10的转录因子。药物-基因相互作用分析确定了46种药物-基因相互作用。体外数据表明，无论是在组织还是细胞系水平，黑色素瘤中CCL4、CCL5、CXCL9和CXCL10的水平均高于正常皮肤组织。
总之，我们鉴定出了与原发性黑色素瘤肿瘤发生和进展相关的4个关键趋化因子成员，这些结果可能有助于阐明黑色素瘤的肿瘤发生机制并促进其治疗。
