成纤维细胞生长因子21对二乙基亚硝胺诱导的L02肝细胞癌变的抑制作用  

Inhibitory effect of fibroblast growth factor-21 on the carcinogenesis of L02 cells induced by diethylnitrosamine

作　　者：李舒欢 韩继武[1] 

机构地区：[1]哈尔滨医科大学附属第四医院消化内科,哈尔滨150001

出　　处：《临床肝胆病杂志》2018年第2期321-326,共6页Journal of Clinical Hepatology

摘　　要：目的探讨成纤维细胞生长因子(FGF)21对二乙基亚硝胺(DEN)诱导的L02肝细胞癌变的抑制作用。方法培养L02肝细胞,分别用不同浓度的DEN(1、10、20、50、100和150μmol/L)作用,MTT法测定DEN对L02肝细胞活力的影响,选择适当的DEN刺激浓度(20μmol/L)进行后续实验。同时,分别采用丙二醛(MDA)及超氧化物歧化酶(SOD)检测试剂盒检测DEN(20μmol/L)作用细胞和正常L02细胞内的MDA及SOD水平。将L02肝细胞分为模型对照组(20μmol/L DEN+PBS处理)、FGF21低剂量组(20μmol/L DEN+1μmol/L FGF21蛋白)和FGF21高剂量组(20μmol/L DEN+2μmol/L FGF21蛋白)。细胞培养12 h后,检测细胞内的MDA及SOD水平。同时采用Real-time PCR和Western Blot检测β-Klotho(KLB)的表达水平,并用Western Blot检测蛋白激酶B(PKB,又称AKT)磷酸化水平。计量资料2组间比较采用t检验,多组间比较采用方差分析,进一步两两比较采用LSD-t检验。结果 DEN作用于L02后细胞内MDA水平明显升高,SOD水平显著下降,差异均有统计学意义(t值分别为9.336和16.281,P值分别为0.011和0.004)。与模型对照组相比,FGF-21低剂量组和FGF-21高剂量组细胞内MDA水平下降、SOD水平升高(P值均<0.05),且FGF21高剂量组细胞内MDA水平显著低于FGF21低剂量组,差异有统计学意义(P=0.030),SOD水平则高于FGF21低剂量组,差异有统计学意义(P=0.042)。FGF21高剂量组与FGF21低剂量组细胞内KLB mRNA水平明显高于模型对照组(P值均<0.001),而FGF21高剂量组KLB mRNA水平高于低剂量组,差异有统计学意义(P<0.001),KLB蛋白表达趋势与KLB mRNA相同。模型对照组中磷酸化AKT(p-AKT)水平高于另外2组,同时FGF21高剂量组p-AKT水平低于FGF21低剂量组,差异均有统计学意义(P值均<0.05)。结论 DEN可增强L02细胞内氧化应激,FGF21通过上调KLB表达,降低AKT磷酸化水平,抑制DEN诱导L02肝细胞氧化应激,从而抑制肝细胞癌变的发生。Objective To investigate the inhibitory effect of fibroblast growth factor-21（FGF-21） on the carcinogenesis of L02 cells induced by diethylnitrosamine（DEN）.Methods L02 cells were cultured and treated with different concentrations of DEN（1,10,20,50,100,and 150 μmol/L）.MTT assay was used to measure the influence of DEN on the viability of L02 cells,and an appropriate stimulation concentration of DEN（20 μmol/L） was selected for further study.The malondialdehyde（MDA） and superoxide dismutase（SOD） detection kits were used to measure the levels of MDA and SOD in L02 cells treated by DEN（20μmol/L） and normal L02 cells.Then L02 cells were divided into model control group（treated with 20μmol/L DEN and PBS）,low-dose FGF-21 group（20 μmol/L DEN ＋ 1 μmol/L FGF-21）,and high-dose FGF-21 group（20 μmol/L DEN ＋ 2 μmol/L FGF-21）.The levels of MDA and SOD were measured after 12 hours of cell culture.Real-time PCR and Western blot were used to measure the expression of βKlotho（KLB）,and Western blot was used to measure the level of phosphorylated protein kinase B（p-AKT）.The t-test was used for comparison of continuous data between two groups; an analysis of variance was used for comparison between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results There was a significant increase in the level of MDA and a significant reduction in the level of SOD after L02 cells were treated with DEN（t = 9.336 and 16.281,P = 0.011 and 0.004）.Compared with the model control group,the low-and high-dose FGF-21 groups had a significant reduction in the level of MDA and a significant increase in the level of SOD（P〈0.05）,and compared with the low-dose FGF-21 group,the high-dose FGF-21 group had a significantly lower level of MDA and a significantly higher level of SOD（P = 0.030 and 0.042）,and there was a significant difference between two groups.The high-and low-dose FGF-21 groups had significantly higher mRNA expres

关 键 词：肝细胞 成纤维细胞生长因子 二乙基亚硝胺 氧化性应激 

分 类 号：R735.7[医药卫生—肿瘤]