CAJ | 학술논문

目的观察表没食子儿茶素没食子酸酯(EGCG)是否具有增强氟尿嘧啶(5-Fu)抑制肝癌Hep3B细胞生长作用,并探讨其机制.方法应用噻唑蓝(MTT)比色法观察肝癌细胞的生存率,应用q值判断药物间的相互作用；Western blot法测定磷酸化乙酰辅酶A羧化酶(p-ACC)和磷酸化蛋白激酶B(p-Akt)蛋白表达.结果与对照组[0.1％二甲基亚砜(DMSO)]比较,工作浓度为5、10、25、50 μmoL/L的EGCG显著抑制Hep3B细胞的生长(P＜0.05).当与30 μmol/L 5-Fu联合用药时,相应的q值分别为1.09、1.27、1.30和1.16,即浓度为5 μmol/L EGCG与5-Fu联合用药表现出两药相加效应,而浓度为10、25、50 μmol/L EGCG与5-Fu联合用药表现出协同效应.EGCG上调Hep3B细胞中p-ACC(Ser79)蛋白表达,下调p-Akt(Thr308)蛋白表达,进一步揭示EGCG与5-Fu联合激活腺苷酸活化蛋白激酶(AMPK)通路,抑制磷酸肌醇3激酶(PI3K)/蛋白激酶B(Akt)通路.这一系列分子活动参与5-Fu协同抗肝癌细胞生长作用.结论 EGCG具有协同增强5-Fu抑制Hep3B细胞生长作用.
목적 관찰표몰식자인다소몰식자산지(EGCG)시부구유증강불뇨밀정(5-Fu)억제간암Hep3B세포생장작용,병탐토기궤제.방법 응용새서람(MTT)비색법관찰간암세포적생존솔,응용q치판단약물간적상호작용；Western blot법측정린산화을선보매A최화매(p-ACC)화린산화단백격매B(p-Akt)단백표체.결과 여대조조[0.1％이갑기아풍(DMSO)]비교,공작농도위5、10、25、50 μmoL/L적EGCG현저억제Hep3B세포적생장(P＜0.05).당여30 μmol/L 5-Fu연합용약시,상응적q치분별위1.09、1.27、1.30화1.16,즉농도위5 μmol/L EGCG여5-Fu연합용약표현출량약상가효응,이농도위10、25、50 μmol/L EGCG여5-Fu연합용약표현출협동효응.EGCG상조Hep3B세포중p-ACC(Ser79)단백표체,하조p-Akt(Thr308)단백표체,진일보게시EGCG여5-Fu연합격활선감산활화단백격매(AMPK)통로,억제린산기순3격매(PI3K)/단백격매B(Akt)통로.저일계렬분자활동삼여5-Fu협동항간암세포생장작용.결론 EGCG구유협동증강5-Fu억제Hep3B세포생장작용.
Objective To investigate whether epigallocatechin-3-gallate (EGCG) enhances 5-fluorouracil (5-Fu)-induced cell growth inhibition in hepatocellular carcinoma cell line Hep3B,and explore its potential mechanisms.Methods 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to evaluate cell viability.The q value was applied to assess the mutual interactions among drugs.Western blotting analysis was performed to detect the phosphor-acetyl-coA carboxylase-ser 79(p-ACC,Ser79) and phosphor-protein kinase B (p-Akt,Thr308) protein expression.Results 5,10,25,and 50 μmol/L EGCG could significantly inhibite the cell growth in Hep3B ceils as compared with the control group (0.1％ DMSO,P ＜0.05).When cells were treated with 30 μmol/L 5-Fu in combination with either 5,10,25,or 50 μmol/L EGCG,the corresponding q value was 1.09,1.27,1.30,and 1.16 respectively.These data suggested that the two drugs had summation action with the combination of 5 μmol/LEGCG and 5-Fu,and had synergic effect with the other three concentrations of EGCG plus 5-Fu.Moreover,the further investigations showed that EGCG up-regulated the expression of p-ACC (Ser79) and downregulated p-Akt (Thr308),suggesting the activation of AMP-activated protein kinase (AMPK) and the inhibition of PI3K/Akt signaling pathway.These observations were involved in the synergic effect.Conclusion EGCG enhances 5-Fu-induced cell growth inhibition of Hep3B cells.