沉默 NEK2 通过调节 Wnt 信号通路抑制非小细胞肺癌的细胞增殖

目的 探讨沉默有丝分裂相关激酶 2 ( never in mitosis related kinase 2, NEK2) 对非小细胞肺癌(non-small cell lung cancer, NSCLC) 细胞生物学行为的影响及其可能机制。 方法 收集 27 对 NSCLC 和邻近的正常组织, 免疫组化检测组织 NEK2 表达, 收集临床病理数据。 生物信息学分析研究 NEK2 在肺腺癌中的表达和预后价值。 通过 NCI-H1299 和 A549 细胞体外转染 siRNA 敲低 NEK2, 通过免疫印迹及 qPCR 实验验证敲低效果, CCK-8 评估细胞增殖能力, 流式细胞术评估细胞凋亡及周期, 蛋白印迹测定 Wnt 信号通路相关蛋白表达。 结果 在肺腺癌组织中的 NEK2 表达显著高于癌旁组织, ⅡB-ⅢB 级组显著高于ⅠA-ⅡA级组, 从 TCGA 数据库获得的数据也显示肺腺癌患者的 NEK2 明显高于相应的对照组, 并与预后不良显著相关。 转染 NEK2 siRNA 显著降低了 NCI-H1299 和 A549 细胞的增殖, 细胞阻滞在 G0/ G1期, 细胞凋亡率增加。 此外, 转染 NEK2 siRNA 的 NCI-H1299 和 A549 细胞中 Wnt 和 β-catenin 的表达水平显著下调, 而 p-GSK3β 的表达水平上调。 结论 siRNA 干扰 NEK2 的表达可抑制非小细胞肺癌细胞的增殖, 其机制可能是通过抑制 Wnt / β-catenin 通路的促进细胞凋亡和周期阻滞。

Silencing NEK2 Inhibits Cell Proliferation of Non-small Cell LungCancer by Regulating Wnt Signaling Pathway

Objective To explore the effect of silencing NEK2 on the biological behavior ofnon-small cell lung cancer (NSCLC) cells and its possible mechanism. Methods A total of 27pairs of NSCLC and adjacent normal tissues were collected, the expression level of NEK2 in thosetissues were detected immunohistochemically, and the clinicopathological data were collected. Bioinformatics methods were performed to analyze the expression level and the prognostic value ofNEK2 in lung adenocarcinoma. NCI-H1299 and A549 cells were transfected with siRNA to knockdown the expression of NEK2 in vitro, and the knockdown efficiency was assessed by Western blotting and qPCR experiments. CCK-8 was used to measure the proliferation rate of cells, and flow cytometry was used to detect the cell apoptosis and cell cycle. Western blotting was used to detect theexpression levels of Wnt signaling pathway related proteins. Results The expression level of NEK2in lung adenocarcinoma tissues was significantly higher than that in the adjacent tissues, and its expression level in the ⅡB-ⅢB group was significantly higher than that in the IA-ⅡA group. The dataobtained from the TCGA database also showed that the expression level of NEK2 in patients withlung adenocarcinoma was significantly higher than that in patients of the corresponding controlgroup, and that its high expression level was significantly associated with a poor prognosis. Transfection of NEK2 siRNA significantly inhibited the proliferation and enhanced the apoptosisof NCI-H1299 and A549 cells, made the cells been arrested in the G0/ G1 phase. In addition, theexpression levels of Wnt and β-catenin were significantly lower in the NCI-H1299 and A549 cellstransfected with NEK2 siRNA, while the expression level of p-GSK3β was higher. Conclusion Silencing of NEK2 by siRNA can inhibit the proliferation of non-small cell lung cancer cells, themechanism may relate to the promotion of cell apoptosis and cell cycle arrest through the inhibition ofthe Wnt / β-catenin pathway.