| 摘要: |
| 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（20 μmol/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 Western blotting 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）， Western blotting 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。 |
| 关键词: 胃癌 极光激酶 A Wnt 信号通路 上皮细胞 - 间充质转化 |
| DOI：10.3969/j.issn.1007-6948.2019.04.001 |
|
| 基金项目:国家自然科学基金（81573737） |
|
| Effect of Aurora Kinase A on Invasion and Migration of Gastric Cancer Cells through Wnt Signaling Pathway |
| WEI Xiao-dong,LIU Xi,TANG Yan-ping |
| Department of Gastroenterology, Tianjin Nankai Hospital, Tianjin (300100), China |
| Abstract: |
| 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 MLN 8237 increased the level of E-cadherin (4 .05± 0.96, P < 0 .05). And the treatment of MLN8238 induced the G 2 /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-egulating Wntsignaling pathway activity. AURKA can be used as a potential target to improve the sensitivity of gastric cancer to chemotherapy. |
| Key words: Gastric cancer Aurora Kinase A |
