CAJ | 학술논문

目的探讨胰岛素因子受体激活后凋亡抑制因子survivin在肝癌细胞株HepG2中的表达变化及其信号传导途径.方法利用实时定量PCR和Western bolt方法,检测HepG2细胞在胰岛素刺激前后survivin的表达变化;在肝癌细胞HepG2中加入Ly294002,特异性阻断磷脂酰肌醇-3羟基激酶(P13K)信号通路,以观察survivin的表达变化与P13K信号传导途径的关系.结果在无血清培养条件下,survivin在HepG2中少量表达;胰岛素能诱导survivin表达,且呈剂量依赖性和时间依赖性:胰岛素浓度在0.26 nmol/L时,survivin的表达为胰岛素诱导前的7倍;当胰岛素浓度达到26 nmol/L时,survivin的表达明显增加,为胰岛素诱导前的35倍(P<0.05).胰岛素诱导HepG2细胞48 h后,survivin的表达也明显增加,为胰岛素诱导前的30倍(P<0.05).Ly294002预处理的HepG2细胞经胰岛素诱导,可阻断survlvin的表达.结论胰岛素通过PISK信号通路诱导凋亡抑制因子survivin的表达,提示通过干扰PI3K/AKt途径中的关键基因可阻断survivin表达,从而促进肝癌细胞凋亡.
목적 탐토이도소인자수체격활후조망억제인자survivin재간암세포주HepG2중적표체변화급기신호전도도경.방법 이용실시정량PCR화Western bolt방법,검측HepG2세포재이도소자격전후survivin적표체변화;재간암세포HepG2중가입Ly294002,특이성조단린지선기순-3간기격매(P13K)신호통로,이관찰survivin적표체변화여P13K신호전도도경적관계.결과 재무혈청배양조건하,survivin재HepG2중소량표체;이도소능유도survivin표체,차정제량의뢰성화시간의뢰성:이도소농도재0.26 nmol/L시,survivin적표체위이도소유도전적7배;당이도소농도체도26 nmol/L시,survivin적표체명현증가,위이도소유도전적35배(P<0.05).이도소유도HepG2세포48 h후,survivin적표체야명현증가,위이도소유도전적30배(P<0.05).Ly294002예처리적HepG2세포경이도소유도,가조단survlvin적표체.결론 이도소통과PISK신호통로유도조망억제인자survivin적표체,제시통과간우PI3K/AKt도경중적관건기인가조단survivin표체,종이촉진간암세포조망.
Objective To determine the expression level changes of survivin, a inhibitor of apeptosis protein, followed by activation of insulin receptors in human hepatocellular carcinoma HepG2 cell line, and to investigate the signalling pathway involved in the regulation. Methods Human hepatocelhlar carcinoma HepG2 cells were treated with insulin alone or pre-treated with LY294002, a specific inhibitor of PI3K signalling pathway, to determine whether blocking PI3K signaling can attenuate the up-regulation of survivin expression. Real time RT-PCR and Western blot analysis were used to measure survivin mRNA and protein changes before and after treatment, respectively. Results Without serum supplement, HepG2 cells expressed a small amount of survivin. Insulin induced survivin expression in a dose- and time-dependent fashion. Survivin expression was blocked ff ceils were pre-treated with LY294002 prior to insulin stimulation. Conclusion Insulin induces survivin expression via PI3K signalling pathway, suggesting that to interfere the key gene in this signalling pathway may block survivin expression, therefore, promoting apeptosis in hepatocellular carcinoma ceils.