Li Yanfei, Yun Xia, Li Jiaojiao, Bai Meilin
Department of Dermatology, People's Hospital of Inner Mongolia Autonomous Region, Hohhot, Inner Mongolia 010017, PR China; Department of Dermatology and STD, The First Hospital of Hohhot, Hohhot, Inner Mongolia 010030, PR China.
Department of Science and Education, The First Hospital of Hohhot, Hohhot, Inner Mongolia 010030, PR China.
Tissue Cell. 2023 Apr;81:102029. doi: 10.1016/j.tice.2023.102029. Epub 2023 Jan 18.
Melanoma progression involves multiple molecular pathways. In this study, we explored the effect of immunoglobulin heavy constant gamma 1 (IGHG1) on melanoma progression.
IGHG1, zinc finger E-box binding homeobox 1 (ZEB1), and cleavage stimulation factor subunit 2 tau variant (CSTF2T) expression levels were measured by quantitative reverse-transcription polymerase chain reaction and western blot. BALB/c nude mice were subcutaneously injected with A375 cells to develop a transplantation tumor model, 4 weeks after which the tumor tissues were collected. Cell proliferation, cell invasion, and cell migration of A375 and SK-MEL-28 cells were measured after gain- and loss-of-function of IGHG1, ZEB1, and CSTF2T. Binding of ZEB1 to the IGHG1 promoter was assayed by chromatin immunoprecipitation and dual-luciferase reporter gene assays, while binding of CSTF2T to ZEB1 mRNA was investigated by RNA immunoprecipitation and RNA pull-down assays.
IGHG1, ZEB1, and CSTF2T were all highly expressed in human melanoma cell lines. In A375 and SK-MEL-28 cells, ZEB1 binds directly to the IGHG1 promoter. ZEB1 silencing reduced IGHG1 expression and melanoma cell proliferative, invasive, and migratory properties, and these cellular effects were nullified by overexpression of IGHG1. CSTF2T binds directly to ZEB1 mRNA. Silencing of CSTF2T diminished ZEB1 expression and restrained melanoma cell proliferative, invasive, and migratory capabilities. The impacts of CSTF2T silencing on melanoma cells were counteracted by concomitant overexpression of ZEB1. CSTF2T knockdown reduced tumor volume and weight in nude mice.
The CSTF2T-ZEB1-IGHG1 axis promotes melanoma cell proliferation and invasion.
黑色素瘤进展涉及多个分子途径。在本研究中，我们探讨了免疫球蛋白重链恒定γ1（IGHG1）对黑色素瘤进展的影响。
通过定量逆转录聚合酶链反应和蛋白质印迹法检测IGHG1、锌指E盒结合同源框1（ZEB1）和切割刺激因子亚基2τ变体（CSTF2T）的表达水平。将A375细胞皮下注射到BALB/c裸鼠体内以建立移植瘤模型，4周后收集肿瘤组织。在IGHG1、ZEB1和CSTF2T功能获得和丧失后，检测A375和SK-MEL-28细胞的细胞增殖、细胞侵袭和细胞迁移。通过染色质免疫沉淀和双荧光素酶报告基因测定法检测ZEB1与IGHG1启动子的结合，同时通过RNA免疫沉淀和RNA下拉测定法研究CSTF2T与ZEB1 mRNA的结合。
IGHG1、ZEB1和CSTF2T在人黑色素瘤细胞系中均高表达。在A375和SK-MEL-28细胞中，ZEB1直接与IGHG1启动子结合。ZEB1沉默降低了IGHG1表达以及黑色素瘤细胞的增殖、侵袭和迁移特性，而IGHG1的过表达消除了这些细胞效应。CSTF2T直接与ZEB1 mRNA结合。CSTF2T沉默减少了ZEB1表达并抑制了黑色素瘤细胞的增殖、侵袭和迁移能力。ZEB1的共过表达抵消了CSTF2T沉默对黑色素瘤细胞的影响。CSTF2T敲低减少了裸鼠的肿瘤体积和重量。
CSTF2T-ZEB1-IGHG1轴促进黑色素瘤细胞增殖和侵袭。
