柴胡皂甙-d对HepG2细胞恶性表型的逆转作用  

Effect of Saikosaponins-d on reversing malignant phenotype of HepG2 cells in vitro

作　　者：祝葆华[1] 蒲荣[1] 张国平[1] 李明意[1] 王兰天[1] 苑进凯[1] 

机构地区：[1]广东医学院第一临床学院外科教研室,东莞523808

出　　处：《中华肝脏病杂志》2011年第10期764-767,共4页Chinese Journal of Hepatology

基　　金：广东省科技计划资助项目（2008B030301016）;东莞市科技计划项目（DK0821）

摘　　要：目的探讨柴胡皂甙-d（SSd）对人肝癌细胞株HepG2细胞恶性表型的逆转作用及其机制。方法常规培养HepG2细胞至对数生长期，四甲基偶氮唑盐（MTT）比色法观察SSd不同浓度、作用不同时间对HepG2细胞增殖情况的影响。实验组选择10mg／L的SSd作用HepG2细胞48h，Giemsa染色法观察细胞形态学改变；化学发光法测定细胞上清液中甲胎蛋白（AFP）浓度；放射免疫法检测细胞上清液中白蛋白浓度；transwell小室实验观察细胞迁移率；RT—PCR检测p27基因mRNA表达。并与空白对照组做比较。数据在多组间比较采用随机分组单因素方差分析，在两组间比较采用t检验。结果10mg／L的SSd作用48h，对HqoG2细胞的生长抑制最明显（F=266．06，P〈0．01）。实验组HepG2细胞形态倾向于正常分化，形态变小、变圆，核变小、变圆或卵圆形，核／质比例缩小。与对照组比较，实验组穿膜细胞数明显减少[（50．60±4．04）个与（41．00±4．64）个，t=-7．32，P〈0．01）；细胞上清液中自蛋白含量明显增多[（24．7±2．8）×10-3g／L与（31．1±4．9）×10-3g／L，t=7．83，P〈0．051；AFP含量明显减少[（118．2±15．6）×10^3μg／L与（70．3士5．7）×10^3μg／L，t=-10．72，P〈0．01]。实验组p27基因mRNA的相对表达量明显多于对照组（t=22．00，P〈0．05）。结论SSd对人肝癌细胞株HepG2细胞恶性表型具有明显的逆转作用，其机制可能与SSd上调HepG2细胞p27基因mRNA表达使其进入分化过程有关。Objective To observe the effect of SSd on reversing the malignant phenotype of HepG2 cells and to investigate its mechanism in order to prove that SSd is a new choice to prevent and treat HCC. Methods HepG2 cells were cultured and treated by different concentrationgs （0 mg/L, 2.5 mg/L, 5.0 mg/ L, 10.0 mg/L and 20.0 mg/L） of SSd for 24 h, and treated by 10 mg/L of SSd for 0 h, 6 h, 12 h, 24 h, 48 h and 72h respectively. The cell inhibition rates were measured by MTT assay. Then cells were treated by 10 mg/ L SSd for 48 hr in experimental group and treated by no SSd as a control, their morphological changes were observed by contrast phase microscope. The concentrations of ALB and AFP in clear supematant liquid of cells were detected by radio-immunity and chemiluminescence. The cell migration rates were observed by transwell method, the relative expression levels of p27 mRNA were messured by RT-PCR. Results The inhibitive effect of 10 mg/L SSd was the most significant among different concentrations （F= 265.06, P〈 0.01 ）. The shape of HepG2 from experimental group turned into small and round, and their volume ratios of nucleus to plasma decreased. ALB in supematant liquid of HepG2 was higher （t=7.83, P〈0.05, and its AFP was lower （t=-10.72, P〈0.01） as compared to control group. Cells migrated were fewer and p27 mRNA expression of HepG2 was higher in experimental group than that in control group （t=22.00,P〈0.05）. Conclusion SSd could reverse the malignant phenotype of HepG2 cells. It was suggested that the up-regulation of p27 mRNA expression play an important role in the differentiation of HepG2 cells treated by SSd.

关 键 词：癌 肝细胞 恶性表型 柴胡皂甙-D P27 

分 类 号：R735.7[医药卫生—肿瘤]