摘要
目的 探讨干扰素-α（IFN-α）通过PI3K/Akt信号通路对肝癌细胞SMCC-7721的影响及相关分子机制.方法 通过MTT法检测IFN-α对SMCC-7721细胞活力的影响;利用流式细胞术检测IFN-α对SMCC-7721细胞周期的影响;Western印迹分析脂酰肌醇3激酶（PI3K）/蛋白激酶B（Akt）通路的激活状况及其下游靶基因糖原合成酶激酶-3β（GSK-3β）、细胞周期蛋白D1（Cyclin Dl）、p21的表达变化,以及C-myc和Survivin的表达变化情况.结果 MTT结果显示IFN-α（5、10、15 U/L）均可显著抑制细胞的活力（P＜0.05）,并且在48 h抑制程度最高,抑制率分别为（10.5±3.1）％、（24.3±4.6）％、（39.4±5.5）％（P＜0.01）.流式细胞仪结果显示：与对照组比较,（5、10、15 U/L） IFN-α可使细胞周期阻滞在G1期,Western印迹结果显示：（10、15 U/L） IFN-α显著下调PI3K表达并抑制GSK-3β磷酸化（P＜0.05）,（5、10、15 U/L） IFN-α均可抑制Akt磷酸化,并下调Cyclin D1量及上调p21的表达（P＜0.05）,（10、15 U/L） IFN-α均可下调C-myc和Survivin的表达（P＜0.05）.结论 IFN-α可抑制肝癌细胞SMCC-7721生长和增殖,阻断PI3 K/Akt信号通路传导,影响Cyclin D1、p21、C-myc、Survivin等分子表达.
Objective This study explored how interferon alpha （IFN-α） influenced liver cancer SMCC-7721 cell via PI3K/Akt signaling pathway.Methods The effect of IFN-α on the viability of SMCC-7721 cell was measured by MTT assay.The effect of IFN-α on SMCC-7721 cell cycle was measured by flow cytometry.The activation of PI3K/Akt signaling pathway and downstream target genes expression level of glycogen synthase kinase 3 beta （GSK-3β）,and cell cycle protein （Cyclin D1）,p21,C-myc,Survivin were assayed by Western blot.Results MTT assay indicated IFN-α （5,10,15 U/L） could inhibit SMCC-7721 cell viability,and the inhibitory rate was the highest at 48 h.IFN-α made SMCC-7721 cell arrested.Western blotting assays presented IFN-α could down-regulate expression of PI3K and suppress the phosphorylation of Akt,and then inhibit phosphorylation of GSK-3β.It could also reduce the expression levels of Cyclin D1,C-myc and Survivin but increase the level of p21.Conclusion These results suggested IFN-α could depress the growth and proliferation of liver cancer SMCC-7721 cell via blocking PI3K/Akt signaling pathway,and the molecular proteins Cyclin D1,p21,C-myc,Survivin were involved.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2015年第36期2960-2963,共4页
National Medical Journal of China
