去甲肾上腺素对人肝癌细胞HepG2和人正常肝细胞L-02增殖、凋亡的影响

摘要

目的 观察去甲肾上腺素(NE)对人肝癌细胞HepG2和人正常肝细胞L-02增殖、凋亡的影响.方法 体外培养HepG2细胞和L-02细胞,分别加入不同浓度NE处理,继续培养,采用MTr法观察细胞增殖情况,HE染色和Annexin V-FITC/PI染色法观察细胞形态学变化,流式细胞术观察细胞凋亡情况.结果 经0.1、1、10、50μmol/L NE处理24、48 h,HepG2细胞OD值显著下降(P均＜0.05),L-02细胞OD值无明显变化.10 μmol/L NE作用于HepG2细胞可见典型的凋亡形态学改变,胞质收缩、深染,核染色质浓缩;L-02细胞在NE作用前后形态学均无明显改变.HepG2细胞加入10 μmol/L NE后部分细胞呈绿色(为早期凋亡表现),加入50 μmol/L NE后部分细胞膜呈绿色,细胞核呈红色(为晚期凋亡表现);L-02细胞均未显示有荧光.0.1、1、10μmol/L NE作用于HepG2细胞24 h后,其凋亡率均显著高于对照组(0μmol/L NE),且呈剂量依赖关系(P均＜0.05);而上述浓度NE作用于L-02细胞24h后,其凋亡率较对照组先增高后降低(P均＜0.05).结论 NE可以抑制HepG2细胞增殖并诱导其凋亡,而对L-02细胞增殖和凋亡没有影响,提示其在特定类型肝癌临床治疗中具有潜在价值.%Objective To observe the effects of norepinephrine (NE) on the proliferation and apoptosis of human hepatocellular carcinoma cell line HepG2 and human normal liver cell line L-02.Methods HepG2 and L-02 cells were cultured in vitro and were treated with different concentrations of NE.Cell proliferation and apoptosis were examined by MTT assay and flow cytometry,respectively.Cell morphology was observed by HE staining and Annexin V-FITC/PI staining.Results After treatment with 0.1,1,10,and 50 μ mol/L NE for 24 and 48 h,the OD value of HepG2 cells decreased significantly (all P ＜ 0.05),while the OD value of L-02 cells had no significant change.HepG2 cells treated with 10 μnol/L NE showed obvious apoptotic morphology:there was contracted and stained cytoplasm,as well as condensed chromatin.L-02 cells had no significant changes in morphology before and after NE treatment.After adding 10 μmol/L NE to HepG2 cells,the cells showed a green color (early apoptosis).After addition of 50 μmol/L NE,some of the cells showed a green membrane and a red nucleus (which is a sign of late apoptosis).No fluorescence was observed in the L-02 cells.The apoptotic rates of HepG2 cells treated with 0.1,1,and 10 μmol/L NE for 24 h were significantly higher than those of the control group (0 μmol/L NE) with a dose-dependent manner (all P ＜ 0.05).However,the apoptosis rate of L-02 cells treated with the above concentrations of NE for 24 h increased first and then decreased (all P ＜ 0.05).Conclusion NE inhibits the proliferation and induces apoptosis of HepG2 cells,but has no effect on the proliferation and apoptosis of L-02 cells,suggesting that it may have potential value in the clinical treatment of specific types of hepatic carcinoma.