Wnt/β-catenin信号通路对非小细胞肺癌NCI-H23细胞干性的调控作用

Wnt/β-catenin signaling pathway regulates stemness of non-small cell lung cancer cell NCI-H23

目的探讨Wnt/β-连环蛋白(β-catenin)信号通路对非小细胞肺癌NCI-H23细胞干性的调控作用。方法以非小细胞肺癌NCI-H23细胞为对象,以糖原合成酶激酶-3β(GSK-3β)抑制剂(CHIR-99021)和β-catenin siRNA为干预剂,分别采用噻唑蓝比色法、体外干细胞成球培养、划痕实验及免疫印迹法检测其增殖能力、体外成球能力和迁移能力方面的改变。结果 CHIR-99021作用24 h后NCI-H23细胞的增殖能力、干细胞成球率均明显提升,与对照组的差异均有统计学意义(P<0.05),划痕愈合时间缩短。CHIR-999021干预后的NCI-H23细胞、NCI-H23干细胞中,GSK-3β磷酸化显著降低,增殖细胞核抗原、CD133、乙醛脱氢酶1A1、Nanog、基质金属蛋白酶-2表达增加。β-catenin siRNA沉默β-catenin后NCI-H23中CD133、乙醛脱氢酶1A1和Nanog表达减少,干细胞成球率也较对照组降低(P<0.05)。结论 Wnt/β-catenin通路参与了非小细胞肺癌NCI-H23细胞干性调节,这对于非小细胞肺癌的防治有一定价值。

Objective To explore the effect of Wnt/β-catenin signaling pathway on stemness maintenance in NCI-H23 cell. Methods The NCI-H23 was treated with the GSK-3β inhibitor （CHIR-99021） or β-catenin siRNA. Then, the MTT assay, stem cell culture, wound healing assay and Western blotting were used to detect proliferation ability, sphering ability and migration ability, re- spectively. Results Compared with control, there were increased proliferation ability and spher rate in NCI-H23 cell after treatment with CHIR-99021 for 24 h （P〈0. 05）. Wound healing showed that the CH-treated cells were also faster than non-treatment. Mean- while, CH-treated cells appeared to exhibit a declined level of p-GSK-3β. The expression of proliferating cell nuclear antigen, CD133, aldehyde dehydrogenase 1A1, Nanog and matrix metalloproteinase-2 showed a significantly enhance in both of NCI-H23 and NCI-H23 stem cell. After silencing β-catenin with β-catenin siRNA, the expression of CD133, ALDH1A1 and Nanog were declined and the sphere rate was decreased comparing with control. Conclusion Wnt/β-catenin signaling pathway involved in regulation of sternness in NCI-H23 cell.