鸦胆子油脂质体对人肝癌细胞株HepG2抑制作用的体内外研究

Suppression of human hepatocellular cancer cell line HepG2 by Brucea liposome in vitroand in vivo

目的通过体外体内实验探讨鸦胆子油脂质体对人肝癌细胞株HepG2的影响及可能机制。方法利用鸦胆子油脂质体高剂量(5mg/L)、低剂量(2.5mg/L)体外作用于人肝癌细胞株HepG2,MTT比色分析法分析HepG2细胞生长抑制程度,流式细胞仪检测不同药物浓度作用下细胞的凋亡率,透射电镜下观察HepG2细胞形态变化。通过腹腔注射荷瘤裸鼠进行体内试验,观察鸦胆子油脂质体小剂量(12.5mg/L)、大剂量(25mg/L)对其肝脏的病理改变;TUNEL法检测肝脏肿瘤组织凋亡细胞。结果 MTT比色分析法实验结果显示鸦胆子油脂质体可明显抑制HepG2细胞增殖;透射电镜下观察5mg/L鸦胆子油脂质体在体外作用于HepG2细胞6h后,可见明显凋亡细胞形态出现。流式细胞仪检测随鸦胆子油脂质体作用强度和时间的增加,细胞抑制率逐步由33.31%提升至90.62%(P<0.01)。鸦胆子油脂质体腹腔注射组较阴性对照组动物模型死亡时平均体重增加,肿瘤转移率下降(P<0.01);TUNEL法亦提示鸦胆子油脂质体组凋亡细胞率增加(P<0.01)。结论鸦胆子油脂质体对HepG2细胞在体内体外具有抑制增殖的作用,并可诱导肿瘤细胞凋亡。

Objective To study the influence of Brucea liposome on human hepatocellular cancer cell line HepG2 and its mechanisms in vitro and in vivo.Methods Morphologic change of HepG2 cells was observed by transmission electron microscopy after treated with Brucea liposome in vitro;cell proliferation was detected by MTT assay after treated with different doses of Brucea liposome.Flow cytometry was used to detect apoptosis rate at different doses.Nude mice were divided into 4 groups randomly: negative control group,low-dose treatment group,large-dose treatment group and positive control group;the animals were raised in similar conditions for 40 days,had the weight and pathologic changes of the liver recorded at the death point.The apoptotic hepatocarcinoma cells were detected by TUNEL technique.Results Apoptotic cells were found with transmission electron microscope after HepG2 cells were treated with Brucea liposome at 5mg/L for 6 hours in vitro.The proliferation of HepG2 was inhibited significantly by Brucea liposome in a time-dependant manner(when the dose was 2.5 mg/L and 5 mg/L)(P<0.01),which was stronger than that by Brucea javanica oil emusion(P<0.05).Compared with the animals in negative control group,the animal models of treatment groups had a greater weight and a lower metastasis rate at the death point(P<0.01).The rate of apoptosis of hepatocellular carcinoma tissues in treatment groups was gradually increased than that in control group(P<0.01);there were significant differences between the treatment groups(P<0.01) with TUNEL technique.Conclusion Brucea liposome can inhibit the proliferation of HepG2 in vitro and in vivo,and it can induce the apoptosis of the cancer cells.