摘要
目的研究羟考酮对肝癌HepG2细胞增殖、凋亡及磷脂酰肌醇-3-激酶/蛋白激酶B(PI3K/Akt)通路的影响。方法体外培养人肝癌HepG2细胞,分别用1、10、20、40、80μg/ml羟考酮对数期生长的肝癌HepG2细胞分别设为OT-1组、OT-10组、OT-20组、OT-40组、OT-80组,其中无任何处理肝癌HepG2细胞为对照组(NC组),采用四甲基偶氮唑蓝(MTT)法检测6组肝癌HepG2细胞增殖情况;采用平板克隆试验法检测肝癌细胞HepG2细胞增殖能力;采用流式细胞仪和AnnexinV-FITC/PI法体外检测肝癌HepG2细胞凋亡率;Western Blot检测肝癌细胞HepG2细胞中凋亡相关cleaved-caspase3、B细胞淋巴瘤-2(Bcl-2)、Bax蛋白表达水平及PI3K/Akt通路相关PI3K、AKT、p-PI3K、p-AKT、磷酸酶及张力蛋白同系物(PTEN)蛋白表达。结果与NC组比较,OT-1组、OT-10组、OT-20组、OT-40组肝癌HepG2细胞增殖抑制率呈时间依赖性和浓度依赖性增加(P<0.05),OT-40组48 h时,肝癌HepG2细胞抑制率为50.27%。与NC组比较,OT-20组、OT-40组肝癌细胞HepG2平板克隆形成率、Bcl-2、p-PI3K、p-AKT蛋白水平依次降低,凋亡率、Bax、cleaved-Caspase3、PTEN蛋白水平增加(P<0.05)。OT-40组与OT-80组,细胞增殖抑制率、平板克隆形成率、凋亡率及各蛋白水平比较,差异均无统计学意义(P>0.05)。结论羟考酮可抑制肝癌HepG2细胞增殖,促进其凋亡,可能与通过上调Bax、cleaved-Caspase3、PTEN,下调Bcl-2蛋白,抑制PI3K/Akt通路活化有关。
Objective To investigate the effects of oxycodone on proliferation,apoptosis and phosphoinositide-3-kinase/protein kinase B(PI3K/Akt)pathway of hepatocellular carcinoma HepG2 cells.Methods Human hepatocellular carcinoma HepG2 cells were cultured in vitro and divided into OT-1 group,OT-10 group,OT-20 group,OT-40 group and OT-80 group after treating with 1μg/ml,10μg/ml,20μg/ml,40μg/ml and 80μg/ml oxycodone,respectively,and the HepG2 cells without any treatment were enrolled as control group.The proliferation status of HepG2 cells was detected by methyl thiazolyl tetrazolium(MTT)assay,the proliferation ability of HepG2 cells was detected by plate cloning assay,and flow cytometry and AnnexinV-FITC/PI were used to detect the apoptotic rate of HepG2 cells in vitro,and Western Blot was used to detect the expression levels of cleaved-caspase 3,B-cell lymphoma-2(Bcl-2),Bax protein and PI3K,AKT,p-PI3K,p-AKT,phosphatase and tensin homologue(PTEN)related with PI3K/Akt pathway in HepG2 cells.Results As compared with that in control group,the inhibition rate of HepG2 cell proliferation in OT-1 group,OT-10 group,OT-20 group and OT-40 group were significantly increased in a time-dependent and concentration-dependent manner(P<0.05),and the inhibition rate at 48 hours in OT-40 group was 50.27%.As compared with those in control group,the HepG2 plate cloning rate,the expression levels of Bcl-2,p-PI3K and p-AKT protein in OT-20 group and OT-40 group were significantly decreased in turn,and apoptotic rate,and the expression levels of Bax,cleaved-Caspase-3 and PTEN protein were significantly increased(P<0.05).There were no significant differences in cell proliferation inhibition rate,plate clone formation rate,apoptosis rate and protein expression levels between OT-40 group and OT-80 group(P>0.05).Conclusion Oxycodone can inhibit the proliferation and promote the apoptosis of HepG2 cells,which may be related to the up-regulation of Bax,cleaved-Caspase-3 and PTEN,as well as the down-regulation of Bcl-2 protein and inhibition of PI3K/Akt pathway activation.
作者
文放放
李熊刚
WEN Fangfang;LI Xionggang(Department of Anesthesiology,Tianmen First People’s Hospital,Hubei,Tianmen 431700,China)
出处
《河北医药》
CAS
2021年第1期30-34,共5页
Hebei Medical Journal
