摘要
目的探讨磷脂酰肌醇3激酶-蛋白激酶B（PI3K-AKT）信号通路对结肠癌细胞SW480的增殖、凋亡及侵袭能力的影响及其可能机制。方法以不同浓度（0、5、10、20、40μmol／L）PI3K-AKT通路特异性抑制剂LY294002作用于SW480细胞，通过四甲基偶氮唑盐法（Mrrr法）检测24、48、72h后细胞活力；以不同浓度（0、10、20、40μmoL／L）LY294002作用于SW480细胞24h，分别用流式细胞法（FCM）检测各组细胞凋亡率，Transwell小室侵袭实验检测各组细胞侵袭能力改变，Western印迹法检测各组细胞血管内皮生长因子（VEGF）、金属基质蛋白酶9（MMP-9）的表达；运用单因素方差分析进行统计学分析。结果MTT法显示除较低浓度组（5μmol／L）外，其余浓度组SW480细胞增殖率均明显低于0μmol／L（对照）组，且有一定的时间一剂量依赖性（均P〈0．05）。流式细胞结果显示，10、20、40μmol／L组凋亡率分别为13．3％±0．9％、30．9％±2．5％、41．2％±4．1％，均明显高于对照组（5．2％±1．8％，均P〈0．05）；细胞侵袭力检测结果显示，10、20、40μmoL／L组穿透膜的细胞数量依次为（87±6）、（65±7）、（46±11）个，除10μmol／L组（P=0．096）外，其余各组均明显低于对照组[（100±10）个，均P〈0．05]。Western印迹法检测各浓度组VEGF、MMP-9表达均少于对照组（均P〈0．05）。结论PI3K-AKT信号通路是结肠癌细胞株SW480增殖、抗凋亡、侵袭转移的重要因素，其机制可能与抑制VEGF、MMP-9表达有关，PI3K-AKT信号通路可能是结肠癌治疗的潜在靶点。
Objective To explore the effects of phosphatidylinositol 3 kinase-protein kinase B （PI3 K-AKT） signal pathway on the proliferation, apoptosis and invasiveness of colon cancer cell line SW480 and explore its possible mechanisms. Methods SW480 cells were cultured with various concentrations （0, 5, 10, 20 and 40 μmol/L） of LY294002 and methyl thiazolyl tetrazolium （MTF） assay was conducted after 24, 48 and 72 hours. SW480 cells were cultured for24 hours with various concentrations （0, 10, 20 and 40 μmol/L） of LY294002. The apoptotic rate was measured by flow cytometry （FCM）. The difference of invasiveness was examined by Transwell invasion test. Western blotting was used to examine the expressions of vascular endothelial growth factor （VEGF） and matrix metalloproteinase-9 （ MMP-9 ）, One-way ANOVA was used for statistical analysis. Results MTF assay showed that the proliferation rate of all SW480 cells except for the lowest concentration group （5 μmol/L） was lower than the control group （0 μmol/L） and a time-dosage dependence existed （all P 〈0. 05）. The results of FCM demonstrated that the apoptotic rates of 10, 20 and 40 μmol/L groups were 13.3% ± 0. 9% , 30. 9% ± 2. 5% and 41. 2% ± 4. 1% respectively, and were all significantly higher than control group （5.2% ± 1.8% , all P 〈0. 05）. The number of cells penetrating through the membrane of 10, 20 and 40 μmol/L groups were 87 ±6, 65 ± 7 and 46± 11 respectively. All invasiveness groups were all lower than control group （ 100 ± 10, all P 〈0. 05） except the 10μmol/L concentration group （P = 0. 096）. Western blotting showed that the expressions of VEGF andMMP-9 were all lower than control group （ all P 〈 0.05）. Conclusions PI3 K-AKT signal pathway plays an important role in the proliferation, apoptosis and inhibition of colonic cancer cells. Its mechanism is probably related with the inhibitions of VEGF and MMP-9. PI3 K-AKT signal pathway may become a potential target for the treatment of colon cancer.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2013年第8期623-626,共4页
National Medical Journal of China
关键词
结肠肿瘤
肿瘤侵润
信号传导
Colonic neoplasms
Neoplasm invasiveness
Signal transduetion
