【摘要】 目的:研究PI3K特异性抑制剂LY294002对人大肠癌细胞系HCT-8生长的影响。方法:将对数生长期的HCT-8细胞分组培养,对照组加入不含LY294002的培养液,实验1～4组加入LY294002,其终浓度分别为5、10、20和40μmol/L,培养24、48、72、96h后,MTT法测定细胞增殖抑制率。取传代培养并贴壁的HCT-8细胞分组培养,对照组常规培养,实验组培养液中加终浓度为10μmol/L的LY294002,培养24、48和72h后流式细胞仪检测细胞周期。取对数生长期的HCT-8细胞,对照组常规培养,实验组培养液中加终浓度为10μmol/L的LY294002,培养24h,培养前后,免疫细胞化学SP法检测P-Akt和NF-κB。结果:与对照组比较,随着LY294002剂量、作用时间的增加,对HCT-8细胞增殖抑制率增大(P<0.01)。10μmol/L的LY294002作用于HCT-8细胞,随着作用时间的延长,G0/G1期细胞逐渐增加,S和G2/M期细胞逐渐减少(P<0.05)。对照组培养前后HCT-8细胞P-Akt和NF-κB蛋白表达无明显变化,10μmol/L的LY294002作用后HCT-8细胞P-Akt和NF-κB蛋白表达降低(P<0.01)。结论:LY294002可能通过抑制P-Akt的表达,从而抑制人大肠癌HCT-8细胞的生长。更多还原

【Abstract】 Aim:To study the effect of LY294002 inhibitor on the proliferation of human colorectal cancer cell line HCT-8.Methods:The prohiferation inhibitory rate of HCT-8 cells treated by LY294002 at doses of 5,10,20,40 μmol/L for 24,48,72 and 96 h,respectively,was evaluated using MTT method.Cell cycle of HCT-8 cells treated by LY294002 at a doses of 10 μmol/L for 24,48,and 72 h,respectively,was detected by flow cytometry.The expression of P-Akt and NF-κB in HCT-8 cells treated by LY294002 at a dose of 10 μmol/L for 24 h was detected by immunohistochemistry.HCT-8 cells treated without LY294002 were the control.Results:The proliferation of HCT-8 cells was inhibited in time-and dose-dependent manner by LY294002.After the treatment by 10 μmol/L LY294002 for 24 h,the proportion of HCF-8 cells in G0/G1 phase was higher(P<0.05),that in S and G2/M phase was lower(P<0.05).The protein expression of P-Akt and NF-κB in HCT-8 cells was inhibited by LY294002(P<0.01).Conclusion:LY294002 can inhibit the expression of P-Akt of human colorectal cancer cell line HCT-8,and inhibit the proliferation of HCT-8.更多还原