极光激酶A通过调控Wnt信号通路影响胃癌细胞增殖和侵袭

Effect of Aurora Kinase A on Invasion and Migration of Gastric Cancer Cells through Wnt Signaling Pathway

目的:研究极光激酶A(AURKA)基因通过调控Wnt信号通路抑制胃癌细胞的增殖和侵袭的作用机制。方法:AURKA特异性抑制剂MLN8237(20μmol/L)处理SGC-7901胃癌细胞系作为实验组,以DMSO处理作为对照组,实时定量PCR(qRT-PCR)检测细胞内AURKA的表达水平,藉此验证该抑制剂效果;同时应用qRT-PCR和Westernblotting方法检测Wnt信号通路关键蛋白β-catenin和EMT相关蛋白N-cadherin、E-cadherin和Twist表达水平,检测细胞周期变化,用Transwell法和平板克隆形成实验检测细胞侵袭和增殖。结果:PCR结构提示对照组细胞内AURKA的表达为1.00±0.13,实验组为0.36±0.09(P<0.01);与对照组相比,实验组SGC-7901细胞内β-catenin(0.41±0.07)(P<0.01)、N-cadherin(0.26±0.08)(P<0.01)、Twist(0.33±0.12)(P<0.01)表达水平降低,E-cadherin(4.05±0.96)表达水平上调(P<0.05),Westernblotting实验的结果与其一致,细胞周期实验结果示细胞出现了明显的G2/M期阻滞(P<0.05);transwell实验结果示实验组(28.33±3.82)穿过基膜的细胞较对照组(83.67±4.28)明显减少(P<0.05);平板克隆实验结果显示实验组克隆形成数(104.67±5.73)较对照组(417.00±7.25)明显减少(P<0.05)。结论:AURKA可以通过下调Wnt信号通路活性抑制细胞侵袭、增殖过程,可作为提高胃癌临床化疗敏感性的潜在靶点。

Objective To investigate the effect of Aurora Kinase A (AURKA) on invasion and migration of gastric cancer cells through Wnt signaling pathway. Methods Gastric cancer cells SGC-7901 were treated by AURKA inhibitor, MLN8237 (20μmol/L). The expression of AURKA,β-catenin, N-cadherin, E-cadherin and Twist was detected by real-time quantitative PCR (qRT-PCR) and Western blotting. Cell invasion abilities and proliferation were detected by Transwell and clone formation assay. Results MLN8237 decreased the expression of AURKA (0.36±0.09, P <0.01),β-catenin (0.41±0.07, P <0.01), N-cadherin (0.26±0.08, P <0.01) and Twist (0.33±0.12, P <0.01). However, the treatment of MLN8237 increased the level of E-cadherin (4.05±0.96, P <0.05). And the treatment of MLN8238 induced the G2/M arrest (P <0.05). The Transwell assay showed that the number of cells passing through basement membrane in the experimental group (28.33±3.82) was significantly lower than that in the control group (83.67±4.28, P <0.05). The plate cloning experiment showed that the number of clones formed in the experimental group (104.67±5.73) was significantly lower than that in the control group (417.00±7.25, P <0.05). Conclusion AURKA could inhibit the invasion and proliferation of glioma cells by down-regulating Wnt signaling pathway activity. AURKA can be used as a potential target to improve the sensitivity of gastric cancer to chemotherapy.