Department of Dermatology, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, China.
Science and Technology Innovation Center, Shandong First Medical University & Shandong Academy of Medical Sciences, Jinan, Shandong, China.
Exp Dermatol. 2023 Jan;32(1):24-29. doi: 10.1111/exd.14677. Epub 2022 Oct 2.
In the past 10 years, the systemic treatment of advanced melanoma has undergone tremendous changes through the development of targeted therapy. However, there is still a long way to go. This study aims to characterize the function and interaction of ITGAX, SERPINB8 and furin in BRAF V600E mutant melanoma. Differentially expressed genes related to BRAF V600E mutation and BRAFi treatment were obtained by analysing GSE141484 and GSE22838. two kinds of BRAFi (Vemurafenib, 10 μM; Dabrafenib, 1 μM) were used to treat A375 and 1205Lu cells, respectively. The expression of ITGAX, SERPINB8 and Furin in A375 and 1205Lu cells was down-regulated by specific siRNAs, and cell proliferation, clone formation and invasion were detected by CCK-8, colony formation and transwell assays. The physical binding of furin and SERPINB8 was detected by immunoprecipitation. BRAFi treatment down-regulated the ITGAX and SERPINB8 expression and did not change furin expression. Knockdown of ITGAX and SERPINB8 both inhibited the proliferation and invasion of A375 and 1205Lu cells. Knocking down SERPINB8 down-regulated the expression of ITGAX. Furin knockdown and inhibitors all up-regulated the protein level of ITGAX. SERPINB8 can physically bind to furin. In summary, SERPINB8 and furin regulate the expression of ITGAX in melanoma cells, and ITGAX significantly promotes the proliferation and invasion of melanoma cells.
在过去的 10 年中，通过靶向治疗的发展，晚期黑色素瘤的系统治疗发生了巨大变化。然而，仍有很长的路要走。本研究旨在研究 ITGAX、SERPINB8 和 furin 在 BRAF V600E 突变黑色素瘤中的功能和相互作用。通过分析 GSE141484 和 GSE22838 获得与 BRAF V600E 突变和 BRAFi 治疗相关的差异表达基因。分别用两种 BRAFi（vemurafenib，10μM；dabrafenib，1μM）处理 A375 和 1205Lu 细胞。用特异性 siRNA 下调 A375 和 1205Lu 细胞中 ITGAX、SERPINB8 和 Furin 的表达，用 CCK-8、集落形成和 Transwell 检测细胞增殖、克隆形成和侵袭。用免疫沉淀检测 furin 和 SERPINB8 的物理结合。BRAFi 处理下调 ITGAX 和 SERPINB8 的表达，而不改变 furin 的表达。敲低 ITGAX 和 SERPINB8 均抑制 A375 和 1205Lu 细胞的增殖和侵袭。敲低 SERPINB8 下调 ITGAX 的表达。Furin 敲低和抑制剂均上调 ITGAX 的蛋白水平。SERPINB8 可与 furin 物理结合。总之，SERPINB8 和 furin 调节黑色素瘤细胞中 ITGAX 的表达，ITGAX 显著促进黑色素瘤细胞的增殖和侵袭。
