Cai Weilin, Chen Minliang
Medical School of Chinese PLA, Beijing, People's Republic of China.
Senior Department of Burns and Plastic Surgery, the Fourth Medical Center of PLA General Hospital, Beijing, People's Republic of China.
J Inflamm Res. 2024 Mar 15;17:1687-1706. doi: 10.2147/JIR.S447934. eCollection 2024.
Tumor growth induces the tumor margin to become a transition zone rich in immune cells. EVPL is a potential prognostic biomarker for melanoma. Melanoma is difficult to cure because of its high metastasis, so it is urgent to find effective genes to inhibit tumor progression and regulate tumor microenvironment.
Firstly, differentially expressed genes (DEGs) among normal skin, nevus and melanoma samples in GSE3189 were screened. Bioinformatics was used to further explore the hub genes and enriched pathways closely related to the inflammatory response of DEGs in melanoma. We selected EVPL, which is associated with the Ras/Raf signaling pathway, for in vitro study. CCK-8, colony formation, wound healing, Transwell and flow cytometry assays were respectively used to evaluate the proliferation, migration, invasion, and apoptosis of cancer cells. Enzyme-linked immunosorbent assay was conducted for the monitoring of changes in the tumor microenvironment. To evaluate the effect of EVPL on macrophage recruitment, we established a co-culture system in a Transwell chamber. The polarization of macrophages was examined after treatment of cells with RAS/ERK signaling inhibitors SCH772984 and sh-EVPL. Additionally, changes in the expression of pathway proteins were measured by Western blot.
Among the screened hub genes, EVPL was associated with the Ras/Raf pathway, a key signaling pathway in melanoma, and may be involved in regulating the inflammatory microenvironment of melanoma. Oe-EVPL was proved to suppress melanoma cell malignant progression. By inhibiting EVPL expression, the inhibitory effects on melanoma progression induced by the addition of SCH772984 were reversed. Furthermore, EVPL was found to inhibit the expression of chemokines, the recruitment of macrophages, and the polarization of macrophages through the Ras/Raf/ERK signaling pathway.
EVPL can inhibit the progression of melanoma through the RAS/ERK signaling pathway, change the inflammatory tumor microenvironment of melanoma, and inhibit the recruitment of macrophages.
肿瘤生长促使肿瘤边缘成为富含免疫细胞的过渡区。EVPL是黑色素瘤潜在的预后生物标志物。黑色素瘤因其高转移性而难以治愈，因此迫切需要找到有效的基因来抑制肿瘤进展并调节肿瘤微环境。
首先，筛选GSE3189中正常皮肤、痣和黑色素瘤样本之间的差异表达基因（DEGs）。利用生物信息学进一步探索与黑色素瘤中DEGs炎症反应密切相关的枢纽基因和富集通路。我们选择与Ras/Raf信号通路相关的EVPL进行体外研究。分别采用CCK-8、集落形成、伤口愈合、Transwell和流式细胞术检测评估癌细胞的增殖、迁移、侵袭和凋亡。进行酶联免疫吸附测定以监测肿瘤微环境的变化。为评估EVPL对巨噬细胞募集的影响，我们在Transwell小室中建立了共培养系统。在用RAS/ERK信号抑制剂SCH772984和sh-EVPL处理细胞后，检测巨噬细胞的极化情况。此外，通过蛋白质印迹法检测通路蛋白表达的变化。
在筛选出的枢纽基因中，EVPL与黑色素瘤的关键信号通路Ras/Raf通路相关，可能参与调节黑色素瘤的炎症微环境。过表达EVPL被证明可抑制黑色素瘤细胞的恶性进展。通过抑制EVPL表达，添加SCH772984对黑色素瘤进展的抑制作用被逆转。此外，发现EVPL通过Ras/Raf/ERK信号通路抑制趋化因子的表达、巨噬细胞的募集以及巨噬细胞的极化。
EVPL可通过RAS/ERK信号通路抑制黑色素瘤的进展，改变黑色素瘤的炎症肿瘤微环境，并抑制巨噬细胞的募集。
