慢病毒介导SOX9下调体外抑制人黑素瘤细胞MV3的转移潜能

Lentivirus Mediated SOX9 Down-regulation Inhibits Metastatic Potential of Melanoma Cells MV3 in Vitro

目的研究慢病毒介导的性别决定区Y框蛋白9(SOX9)下调对黑素瘤细胞转移潜能的影响。方法在人黑素瘤细胞MV3中转染SOX9 shRNA慢病毒和阴性对照慢病毒,Realtime PCR和Western blot检测干扰效果。CCK-8法检测各组细胞增殖能力变化,克隆形成实验测定各组细胞克隆形成能力,划痕愈合实验检测各组细胞运动能力,Transwell小室检测各组细胞侵袭和迁移能力,Western blot检测各组细胞中EMT相关蛋白Vimentin、E-cadherin和侵袭迁移相关蛋白MMP-2、MMP-9水平变化。结果与阴性对照慢病毒转染的细胞比较,SOX9 shRNA慢病毒转染后细胞中的SOX9 mRNA和蛋白水平降低,同时细胞增殖和克隆形成能力降低,细胞运动、侵袭和迁移能力也下降,细胞中MMP-2、Vimentin、MMP-9蛋白水平降低,E-cadherin蛋白水平升高。结论慢病毒介导的SOX9下调具有体外抑制黑素瘤细胞转移潜能的作用。

Objective To study the effect of lentivirus mediated SOX9 down-regulation on the metastatic potential of melanoma cells.Methods SOX9 shRNA lentivirus and negative control lentivirus were transfected into human melanoma MV3 cells.Realtime PCR and Western blot were used to detect the interference effect.The cell proliferation ability of each group was detected by CCK-8.Colony formation assay was used to determine the colony forming ability of each group,and scratch healing test was used to detect cell motility in each group.The cell invasion and migration ability of each group were detected by Transwell chamber.The levels of EMT-related proteins Vimentin,E-cadherin,MMP-2 and MMP-9 were detected by Western blot.Results Compared with cells transfected with negative control lentivirus,the levels of SOX9 mRNA and protein in SOX9 shRNA lentivirus transfected cells were decreased after transfection.In the meantime,cell proliferation and colony formation ability was decreased,and cell motility,invasion and migration ability was also declined.In addition,the levels of MMP-2,Vimentin and MMP-9 protein were decreased,and the level of E-cadherin protein was increased.Conclusion Lentivirus mediated SOX9 downregulation is able to inhibit the metastatic potential of melanoma cells in vitro.