LY294002和PD98059对结直肠癌相关血管内皮细胞增殖和凋亡的作用

Role of LY294002 and PD98059 in Proliferation and Apoptosis of Colorectal Carcinoma Related Vascular Endothelial Cells

目的探讨LY294002和PD98059对结直肠癌相关血管内皮细胞增殖和凋亡的作用及其可能机制。方法用结直肠癌LOVO细胞培养上清诱导人脐静脉内皮细胞产生肿瘤相关内皮细胞,设置LY294002组和PD98059组,分别用含LY294002(10μmol/L)和PD98059(10μmol/L)的完全培养液处理肿瘤相关血管内皮细胞;设置空白对照组和二甲基亚砜(DMSO)对照组,分别用完全培养液及含0. 10%DMSO完全培养液培养细胞;采用细胞计数法检测各组细胞增殖,Annexin V-FITC/PI法检测各组细胞凋亡率,比较不同组别细胞增殖和凋亡情况。结果第1天至第5天,空白组细胞数目呈"S"型曲线增长;随时间的延长,LY294002及PD98059对肿瘤相关内皮细胞的增殖的抑制作用趋于明显,各组间、不同时点间、组间和时点间交互作用比较差异均有统计学意义(P <0. 01)。LY294002组、PD98059组凋亡率高于空白对照组[(13. 2±2. 8)%、(15. 1±3. 4)%比(3. 3±1. 7)%](P <0. 05),LY294002组和PD98059组内皮细胞凋亡率比较差异无统计学意义(P=0. 372)。结论 LY294002抑制磷脂酰肌醇-3-激酶/蛋白激酶B和PD98059抑制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均能抑制结直肠癌相关内皮细胞的增殖和促进结直肠癌相关内皮细胞的凋亡。

Objective To investigate the effect and possible mechanism of LY294002 and PD98059 on the proliferation and apoptosis of tumor related vascular endothelial cells. Methods Human umbilical vein endothelial cells were induced to produce tumor associated endothelial cells by being treated with supernatant of colorectal carcinoma LOVO cells.Two experimental groups which were treated with complete medium containing signal pathway inhibitor LY294002（10 μmol/L） and signaling pathway inhibitor PD98059（10 μmol/L） respectively were set up,and two control groups（blank control group and DMSO group） which were treated with complete culture and complete culture containing 0.1% dimethyl sulfoxide（DMSO） respectively were set up.Cell proliferation was assessed by cell counting method,and apoptotic cell death was detected by Annexin V-FITC/PI method.Cell proliferation and apoptosis were compared between different groups. Results From the first to the fifth day,the number of cells in the blank group increased in ＂S＂ curve,and the inhibitory effect of LY294002 and PD98059 on the proliferation of tumor-associated endothelial cells tended to be obvious with the extension of time.The apoptotic rate of LY294002 and PD98059 group was higher than that of blank control group [（13.2±2.8）%,（15.1± 3.4）% vs （3.3±1.7）%]（ P 〈0.05）.There was no significant difference in the apoptotic rate of endothelial cells between the LY294002 and PD98059 group（ P =0.372）. Conclusion Inhibition of phosphatidylinositol 3-kinase/protein kinase B signaling pathway by LY294002 and mitogen extracellular signal-regulated kinase/extracellular signal-regulated kinase signaling pathway by PD98059 can both inhibit the proliferation of colorectal cancer related endothelial cells and promote the apoptosis of colorectal cancer related endothelial cells.