NS5ATP9对饥饿诱导的肝母细胞瘤HepG2细胞凋亡的作用

Effects of NS5ATP9 on cell apoptosis of HepG2 cell

目的观察NS5ATP9对饥饿诱导肝母细胞瘤HepG2细胞凋亡的作用随时间的变化。方法将NS5ATP9的表达载体、siRNA小分子干扰及其各自的对照分别转染HepG2细胞48小时后,分别用EBSS饥饿细胞12小时、24小时、36小时和48小时后,利用Annexin V/7-AAD流式细胞术检测细胞凋亡率的变化。同时利用化学发光法检测胱冬肽酶-3/7(caspase-3/7)的活性变化,观察饥饿不同时间点NS5ATP9对细胞凋亡的作用变化。结果①与对照组相比,NS5ATP9过表达组在HepG2饥饿24小时、36小时和48小时后,总凋亡率显著减低(t=17.132、9.138、17.318,P=0.003、0.012、0.003);12小时、36小时和48小时caspase-3/7酶活性显著减低(t=7.637、6.944、13.490,P=0.017、0.02、0.005),二者均以48小时最显著。②与对照组相比,si-NS5APT9转染组在HepG2饥饿36小时和48小时总凋亡率显著增加(t=-5.064、-4.342,P=0.034、0.049),12小时、36小时和48小时caspase-3/7酶活性显著增加(t=-8.837、-7.469、-18.630,P=0.013、0.017、0.003),二者均以48小时最显著。结论在HepG2细胞中NS5ATP9能显著抑制饥饿诱导的细胞凋亡,随着饥饿时间的延长,NS5ATP9对细胞凋亡率的抑制越明显,以48小时最为显著。

Objective To observe the effects of NS5ATP9 on cell apoptosis of HepG2 cells changes with the different starvation time. Methods NS5ATP9 expression vector, small molecules of NS5ATP9 siRNA interference and their respective control were transiently transfected into HepG2 cells for 48 hours, respectively. The cells were then starvated with EBSS for 12, 24, 36 and 48 hours. Part cells were used to observe cell apoptosis changes staining by Annexin V / 7-AAD. Part cells were used to detected caspase-3/7 activities. Results （~）Compared with control group, the apoptosis rate of HepG2 in NS5ATP9 overexpressed group reduced at 24, 36 and 48 hours （t = 17.132, 9.138, 17.318; P = 0.003,012, 0.003）, and the caspase-3/7 activities reduced at 12, 36 and 48 hours （t = 7.637, 6.944, 13.490; P = 0.017, 0.02, 0.005）. The differences were most significant at 48 hours. （~）Compared with control group, the apoptosis rate of HepG2 in si- NS5APT9 transfected group increased at 36 and 48 hours （t = -5.064, -4.342; P = 0.034, 0.049）, and the caspase-3/7 activities increased at 12, 36, and 48 hours （t = -8.837, -7.469, -18.630; P = 0.013, 0.017, 0.003）. The differences were most significant at 48 hours. Conclusion NS5ATP9 in HepG2 cells can significantly inhibit the apoptosis induced by starvation; with the starvated time prolonged, the effect of NS5ATP9 inhibiting cell apoptosis becomes more obvious and the difference was most significant at 48 hours.