摘要
目的:探索干扰Bmi-1基因后对其可能的下游基因Akt/PKB活性和P16INK4a基因表达的影响及对肿瘤细胞增殖和细胞衰老的作用.方法:用siRNA技术干扰Bmi-1表达后,运用Western blot检测Bmi-1蛋白及相关蛋白pAkt、Akt和P16INK4a的表达,同时进行SA-β-Gal染色检测细胞衰老,软琼脂克隆形成实验检测细胞的增殖能力.结果:转染Bmi-1 i质粒组平均细胞衰老率28%±3.5%,而对照Ctrli组为16%±2.7%,有明显统计学差异(P<0.01).转染Bmi-1 i质粒组细胞平均克隆形成数为3.4±1.4个,而对照Ctrli组为11±2.3个,两组比较有明显的统计学差异(P<0.01).Bmi-1 i组较Ctrli组Bmi-1和pAkt蛋白表达明显下降,而P16INK4a蛋白表达升高.结论:干扰Bmi-1可以通过降低Akt/PKB活性和上调P16INK4a蛋白表达,促进肿瘤细胞衰老并减弱肿瘤细胞的增殖能力.
AIM： To explore the effect of Bmi-1 knock-down on Akt/PKB activity, P16INK4a expression, cell proliferation and cell senescence. METHODS： Bmi-1 expression in AGS was down-regulated using SiRNA approach; Bmi-1 protein and related proteins （pAkt, Akt, P16INK4a） were detected by Western blot. SA- β-Gal activity assay was applied for detection of the senescent cells, and soft-agar growth assay was used to detect the clone formation RESULTS： In the SA-β-Gal activity assay, the average senescent cell rate in Bmi-1 i group was28% ± 3.5%, compared with 16% ± 2.7% in Ctrl i group （P 〈 0.01）. In the soft-agar growth assay, the average clone formation number was 3.4 ±1.4, compared with 11 ± 2.3 in Ctrl i group （P 〈 0.01）. The Bmi-1 protein level was down-regulated significantly in the Bmi-1 i group, Akt activity was down-regulated and P16INK4a protein was up-regulated. CONCLUSION： Bmi-1 knock-down may promote cell senescence and inhibit cell proliferation via down-regulating Akt/PKB activity and up-regulating P16INK4a.
出处
《世界华人消化杂志》
CAS
北大核心
2009年第14期1390-1393,共4页
World Chinese Journal of Digestology
