抑制磷脂酰肌醇3激酶／丝氨酸苏氨酸蛋白激酶和丝裂原细胞外信号调节激酶／细胞外信号调节激酶信号通路对结肠癌血管内皮细胞功能的影响

Inhibitotory effects of phosphatidyllnositol 3 - kinase/serine/threonine protein kinase and mitogen extracellular signal - regulated kinase/extracellular signal - regulated kinase signaling pathway on function of vascular endothelial cells in colon cancer

目的观察磷脂酰肌醇3激酶（P13K）／丝氨酸苏氨酸蛋白激酶（Akt）及丝裂原细胞外信号调节激酶（MEK）!细胞外信号调节激酶（ERK）信号通路在结肠癌血管形成过程中对结肠癌相关血管内皮细胞（CCRVEC）功能的影响。方法用不同浓度（2．5、7．5、15．0μmol／L）的信号通路抑制剂分别干预CCRVEC的P13K／Akt和MEK／ERK信号通路，通过细胞计数法、细胞划痕、定向迁移、Transwell迁移、侵袭和管道形成实验检测不同的信号通路对CCRVEC的增殖、迁移、侵袭与管道形成能力的影响。结果抑制CCRVEC的P13K／Akt和MEK／ERK信号通路后，其增殖、迁移、侵袭及管道形成能力均受抑制，且该效应随着抑制剂浓度增高而呈增强趋势；15μmol／L的P13K／Akt和MEK／ERK信号通路抑制剂分别处理CCRVEC6h后，其管道形成长度为（1．28±0．13）mm和（1．87±0．13）mln，明显短于对照组[（4．07±0．26）mln，P〈0．05]；24h后，侵袭细胞数为（207．38±24．29）、（264．02±15．54）个，少于对照组[（475．81±16．66）个，P〈0．05]；细胞迁移数分别为（229．50±20．82）、（271．12±16．10）、（472．33±11．83）个（P〈0．05）；30h后，各组间细胞迁移距离差异有统计学意义（P〈0．05）；抑制剂处理6d后细胞增殖数分别为（56．29±3．85）、（46．23±3．31）和（91．23±4．82）个／孔（P〈0．05）；相同的抑制剂浓度抑制P13K／Akt对细胞迁移、侵袭和管道形成抑制作用较MEK／ERK通路抑制效应更明显，但MEK／ERK信号通路对CCRVEC增殖的影响较P13K／Akt信号通路明显（P〈0．05）。结论P13K／Akt和MEK／ERK信号通路均参与结肠癌血管形成中的增殖、迁移、侵袭及管道形成过程；与P13K／Akt信号通路主要影响CCRVEC迁移、侵袭与管道形成等过程不同，MEK／ERK信号通路尽管也影响CCRVEC迁移、侵袭及管道形成，但对增殖的影响则更明显。

Objective To observe the effect of phosphatidylinositol 3 - kinase （PI3K）/serine/ threonine protein kinase （Akt） and mitogen extracellular signal -regulated kinase （MEK）/extracellular signal - regulated kinase （ERK） signaling pathway on colon cancer - related vascular endothelial ceils （CCRVECs） function in the process of the tumor angiogenesis in colon cancer. Methods CCRVECs were treated with different concentrations （2. 5, 7. 5, 15. 0 μmol/L） inhibitors of PI3K/Akt and MEK/ERK signal pathway. The proliferation of CCRVECs was measured by cell counting. The cell migration and inva- sion ability was tested by cell scratch, orientation migration, Transwell assays, respectively, simultaneous- ly, tube formation technique was also performed to detect the tube formation on CCRVECs. Results In CCRVECs, following blocking PI3K/Akt and MEK/ERK signal pathway with special inhibitor, the ability of proliferation, migration and invasion was inhibited. Moreover, the inhibiting effect was increased with the increase of the concentration. After CCVECs were exposed to 15 μmol/L of PI3K/Akt and MEK/ERK inhibitors for 6 h, the length of tube formation in three groups was （ 1.28 ± O. 13 ） mm, （ 1.87 ±O. 13 ） mm and （4. 07± 0. 26 ） mm respectively, and the length in experiment groups was decreased notably as compared with control group （P 〈0. 05 ）. At 24 h after treatment with special inhibitor, the number of in- vasive ceils in control group was 475.81 ± 16. 66, and that in treated groups was 207.38 ± 24. 29 and 264.02 ± 15.54 respectively, with the difference being significant between experiment groups and the con- trol group （ P 〈 0.05 ）. After treatment for 30 h, the cell migration distance showed significant difference among the groups （P 〈 0.05 ）. After CCVECs were exposed to inhibitor for 6 days, the number of proliter- ation （：ells in each well in treated groups and control group was 56. 29 ±3. 85, 46. 23 ±3. 31 and 91.23 ± 4.82 rspectively with the difference being significant among the groups （ P 〈 0. 05 ）. The inhibition effect of PI3K/Akt signal inhibitor was superior to that of MEK/ERK inhibitor in cell migration, invasion, tube formation ability at the same concentration, except cell proliferation ability. Conclusion PI3K/Akt and MEK/ERK signaling pathway partieipate in the process of the cell proliferation, migration, invasion and tube formation of CCVECs in eolon cancer. Unlike PI3K/Akt signaling pathway having main effects on migration, invasion and tune formation of CCVECs, MEK/ERK signaling pathway has more significant influence on cell proliferation of CCVECs.