Institute of Dermatology, Chinese Academy of Medical Sciences, Nanjing, P. R. China.
Department of Dermatology, Xiangya Hospital, Central South University, Changsha, P. R. China.
PLoS One. 2018 Apr 23;13(4):e0195610. doi: 10.1371/journal.pone.0195610. eCollection 2018.
We previously identified ovostatin 2 (OVOS2) as a new candidate gene for cutaneous malignant melanoma (CMM) in a Chinese population. In this study, we aimed to investigate the exact role of OVOS2 in cell proliferation, invasion, and tumorigenesis of melanoma A375 cells.
The downregulation of OVOS2 expression was performed using lentiviral vectors with specific shRNA. The effects of OVOS2 expression on cell proliferation, cell cycle, cell migration, cell invasion, and potential of tumorigenesis were further investigated.
The downregulation of OVOS2 significantly suppressed the proliferation of A375 cells and led to a G2/M phase block. The transwell cell migration assay showed that the reduced expression of OVOS2 also significantly inhibited the transmigration of A375 cells. The western blot results showed downregulated expression of p-FAK, p-AKT, and p-ERK. This was accompanied by the upregulated epithelial phenotypes E-cadherin and β-catenin, and downregulated expression of mesenchymal phenotype N-cadherin after OVOS2 knockdown. The transplantation tumor experiment in BALB/C nude mouse showed that after an observation period of 32 days, the growth speed and weight of the transplanted tumors were significantly suppressed in the BALB/c nude mice subcutaneously injected with OVOS2 knocked-down A375 cells.
The inhibition of OVOS2 had significant suppressive effects on the proliferation, motility, and migration capabilities of A375 cells, suggesting a crucial promotive role of OVOS2 in the pathogenesis and progression of CMM. The involved mechanisms are at least partly associated with the overactivation of FAK/MAPK/ERK and FAK/PI3K/AKT signals.
我们之前在中国人群中发现卵母细胞抑制素 2(OVOS2)是皮肤恶性黑色素瘤(CMM)的一个新候选基因。在这项研究中,我们旨在研究 OVOS2 在黑色素瘤 A375 细胞增殖、侵袭和致瘤性的确切作用。
使用具有特异性 shRNA 的慢病毒载体下调 OVOS2 表达。进一步研究 OVOS2 表达对细胞增殖、细胞周期、细胞迁移、细胞侵袭和潜在致瘤性的影响。
下调 OVOS2 显著抑制了 A375 细胞的增殖,并导致 G2/M 期阻滞。Transwell 细胞迁移实验表明,OVOS2 表达减少也显著抑制了 A375 细胞的迁移。Western blot 结果显示,下调 OVOS2 表达后,p-FAK、p-AKT 和 p-ERK 的表达水平降低。同时,上皮表型 E-钙黏蛋白和β-连环蛋白表达上调,间充质表型 N-钙黏蛋白表达下调。在 BALB/C 裸鼠移植瘤实验中,观察 32 天后,皮下注射 OVOS2 敲低 A375 细胞的 BALB/c 裸鼠中,移植瘤的生长速度和重量明显受到抑制。
OVOS2 的抑制对 A375 细胞的增殖、运动和迁移能力具有显著的抑制作用,提示 OVOS2 在 CMM 的发病机制和进展中具有重要的促进作用。涉及的机制至少部分与 FAK/MAPK/ERK 和 FAK/PI3K/AKT 信号的过度激活有关。