白眉蝮蛇去整合素Adinbitor对Akt信号转导通路及对SSMC7721细胞增殖、迁移和凋亡的影响

研究白眉蝮蛇去整合素adinbitor对蛋白激酶B(Akt)通路信号分子的影响及对SSMC7721细胞增殖、迁移及凋亡的影响.采用MTT法检测adinbitor对SSMC7721细胞增殖的作用；Hoechst33258试剂盒检测adinbitor对SSMC7721凋亡的影响; Transwell 检测adinbitor对SSMC7721细胞迁移的作用；Western印迹检测adinbitor对信号分子Akt及磷酸化蛋白激酶B（p-Akt）、核因子κB（NF-κB）的抑制蛋白IκB-α及NF-κBp65的影响；分光光度法检测其对半胱天冬蛋白酶-3 （caspase -3）活性的影响.结果显示,adinbitor可显著抑制SSMC7721细胞的迁移和增殖(与对照组比较，P＜005)，促进凋亡. 在浓度梯度adinbitor作用下，Akt 表达量基本不变，但其磷酸化受到抑制.细胞浆内IκB α表达增加，细胞核内NF-κBp65表达减少，caspase-3活化倍数平均在2.24～3.85之间，以上作用均呈现剂量依赖性.结果说明，adinbitor可通过抑制Akt相关信号转导分子的作用而抑制SSMC7721细胞增殖和迁移，并促进其凋亡.

Effects of a Disintegrin, Adinbitor, from Gloydius blomhoffi brevicaudus on Akt Signal Pathway and Proliferation, Migration and Apoptosis of SSMC7721 Cells

o explore the effects of adinbitor, a disintegrin derived from Gloydius blomhoffi brevicaudus on Akt pathway and correlative biological functions including proliferation, apoptosis and migration of SSMC7721 cells. MTT assay was used to detect the effect of adinbitor on proliferation. SSMC7721 cells were incubated with different concentrations of adinbitor and stained by fluorescence dye Hoechst 33258 to observe apoptosis. Transwell was used to observe the effect of adinbitor on cell migration. Western blotting was utilized to detect the expression of Akt, p-Akt, IκB-α and RelA（p65）in SSMC7721 cells, which treated with adinbitor. Spectrophotometric method was selected to explore the activity of caspase-3. The results showed that adinbitor could inhibit cell proliferation and enhance cell apoptosis. The number of migrated cells decreased significantly. Adinbitor inhibited the activation of Akt, increased the expression of IκB-α in cytoplasm. The expression of NF-κBp65 in nuclei was decreased accordingly. The activity of caspase 3 was increased remarkably. These results suggested that adinbitor inhibited Akt pathway in a dose dependent manner, so a series of biological function including invasion and apoptosis could be inhibited by adinbitor.