IFI30调控NF-κB信号通路对胶质母细胞瘤细胞增殖和迁移侵袭的影响

Effects of IFI30 on the proliferation, migration and invasion of glioblastoma cells through activating NF-κB signaling pathway

目的 探讨干扰素γ诱导蛋白30(IFI30)对胶质母细胞瘤(GBM)细胞增殖和迁移侵袭的影响及其分子机制。方法 利用GEPIA网站分析GBM组织的IFI30水平,实时荧光定量PCR(qPCR)检测GBM细胞的IFI30水平。采用脂质体法向U251细胞转染靶向IFI30的小干扰RNA序列(IFI30 siRNA组)或无义序列(NC组),另采用核因子(NF)-κΒ激活剂1(Act1)处理IFI30-siRNA转染的细胞(IFI30 siRNA+Act1组),qPCR和Western blotting检测IFI30以及NF-κB p50亚基和抑制蛋白-α(IκB-α)的磷酸化。结果 IFI30在GBM组织表达上调(P<0.05),且IFI30高表达GBM患者的总生存期短于IFI30低表达患者(P<0.05)。N299、U87、A172和U251细胞的IFI30的相对表达水平高于NHA细胞(P<0.05)。相较于NC组,IFI30 siRNA组U251细胞的IkBα磷酸化水平升高,而p50磷酸化表达水平降低,差异有统计学意义(P<0.05);相较于IFI30 siRNA组,IFI30 siRNA+Act1组U251细胞的IkBα磷酸化水平降低,而p50的磷酸化水平升高,差异有统计学意义(P<0.05)。IFI30 siRNA组较NC组的U251细胞增殖活力减弱,侵袭数量和划痕愈合率减少,而IFI30 siRNA+Act1组较IFI30 siRNA组的U251细胞增殖活力增强,侵袭数量和划痕愈合率增多,差异有统计学意义(P<0.05)。结论 IFI30通过激活NF-κB信号通路在GBM中发挥致癌作用,显示其作为GBM潜在治疗靶点的可能性。

Objective To explore the effect of interferon-gamma-inducible protein 30(IFI30) on the proliferation, migration and invasion of glioblastoma cells as well as its molecular mechanism. Methods GEPIA was performed to analysis IFI30 expression in glioblastoma tissues, and real-time fluorescent quantitative PCR(qPCR) was used to detect the levels of IFI30 in GBM cells. Liposome method was used to transfect small interfering RNA sequence targeting IFI30(IFI30 siRNA group) or nonsense sequence(NC group) into U251 cells, and nuclear factor(NF)-κΒ Activator 1(Act1) was used to treat cells transfected with IFI30 siRNA(IFI30 siRNA+Act1 group). Then, qPCR and Western blotting were applied to detect levels of IFI30 and phosphorylation of nuclear NF-κΒ inhibitory protein IκB-α and NF-κΒ p50 subunit. Results IFI30 was up-regulated in GBM tissues(P<0.05), and the overall survival of GBM patients with high IFI30 expression was shorter than that of patients with low IFI30 expression(P<0.05). Relative expression level of IFI30 in N299, U87, A172 and U251 cells was higher than that in NHA cells(P<0.05). Compared with NC group, the phosphorylated level of IkBα increased, while the phosphorylated level of p50 decreased in IFI30 siRNA group(P<0.05). Compared with IFI30 siRNA group, the phosphorylated level of IkBα decreased, while the phosphorylation level of p50 increased in IFI30 siRNA+Act1 group(P<0.05). Compared with NC group, the proliferation activity of U251 cells in IFI30 siRNA group was weaker, and the number of invasions and the rate of scratch healing were reduced(P<0.05). The proliferation activity of U251 cells in IFI30 siRNA+Act1 group was increased, and the number of invasions and the rate of scratch healing were increased in comparison with IFI30 siRNA group(P<0.05). Conclusion IFI30 functions to be oncogenic in glioblastoma through activating NF-κB signaling pathway, indicating that IFI30 may be a potential therapeutic target in glioblastoma.