血管生成素2经PI3K/Akt途径对HCT-8细胞增殖的促进作用  

Promoting effects of Ang-2 through PI3K/Akt pathways on proliferation of HCT-8 cells

作　　者：张继红[1] 王红[2] 任立群[3] 李相军[3] 王彩芹[1] 姜晶[1] 王倩 温春阳[4] 

机构地区：[1]北华大学附属医院病理科,吉林吉林132011 [2]吉林职工医科大学实训中心,吉林吉林132011 [3]吉林大学药学院实验药理与毒理学教研室,吉林长春130021 [4]北华大学医学部,吉林吉林132001

出　　处：《吉林大学学报（医学版）》2011年第6期1057-1061,共5页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅白求恩医学专项基金资助课题(200705401)

摘　　要：目的:研究外源性血管生成素2(Ang-2)蛋白对结肠癌细胞株(HCT-8)中Tie-2、1-磷脂酰肌醇3-激酶(PI3K)、Akt基因及蛋白表达的影响,探讨其与PI3K/Akt通路的关系。方法:用不同浓度的Ang-2蛋白(0、0.2、0.4、0.8、1.2、2.0、3.0mg·L-1)作用于HCT-8细胞,用MTT法检测细胞增殖活性,根据实验结果选定Ang-2蛋白的后续实验浓度。细胞分为正常对照组(C组)、无血清培养液组(SD组)、Ang-2组(SA2组)及LY294002组(SA2L组),应用RT-PCR及Western blotting技术分析Tie-2、PI3K、Akt基因及蛋白的变化。结果:加入不同浓度的外源性Ang-2蛋白,HCT-8细胞均有不同程度的增殖,当Ang-2的浓度为2mg·L-1时,细胞的增殖力最强。SA2组中Tie-2、PI3K及Akt 3种基因及蛋白的表达均较SD组增强(Tie-2:均P<0.01;PI3K:P<0.01及P>0.05;Akt:均P<0.01),而在SA2L组中3种基因及蛋白的表达均较SA2组减弱(Tie-2:P<0.05及P<0.01;PI3K:P<0.05及P<0.01;Akt:P<0.01及P<0.05)。结论:Ang-2蛋白在结肠癌细胞中有促增殖作用,且其作用机制与Ang-2/Tie-2/PI3K/Akt调节的通路有关,应用LY294002可抑制该通路,实现抗肿瘤作用。Objective To observe the effects of angiopoietin-2(Ang-2) on the expressions of Tie-2,PI3K,Akt gene and protein in the colon cancer cell line(HCT-8) and analyze the relationship between Ang-2 and PI3K/Akt.Methods Different doses of Ang-2(0,0.2,0.4,0.8,1.2,2.0,and 3.0 mg·L-1) were added into the DMEM medium of HCT-8 cell line and the effective stimulus concentration of Ang-2 on HCT-8 was selected by MTT.The cells were divided into four groups: normal control group(C group),serum-free culture medium group(SD group),Ang-2 group(SA2 group) and LY294002 group(SA2L group),the gene and protein expressions of Tie-2,PI3K and Akt were detected by RT-PCR and Western blotting.Results Different concentrations of exogenous Ang-2 showed various proliferation effects on the HCT-8 and the most effective dose of Ang-2 was 2 mg·L-1.Compared with SD group,the expressions of Tie-2,PI3K and Akt gene and protein in SA2 group were increased significantly(Tie-2: all P<0.01;PI3K: P<0.01 or P> 0.05;Akt: P<0.01);compared with SA2 group,the expressions of three genes and proteins in SA2L group were decreased significantly(Tie-2: P<0.05 or P<0.01;PI3K: P<0.05 or P<0.01;Akt: P<0.01 and P<0.05).Conclusion Ang-2 can promote the proliferation of the colon cancer cells through the Ang-2/Tie-2/PI3K/Akt pathway which can be inhibited by LY294002.

关 键 词：血管生成素2 结肠癌细胞株 增殖 受体 TIE-2 1-磷脂酰肌醇3-激酶 癌基因蛋白质v-Akt 

分 类 号：R735[医药卫生—肿瘤]