南方医科大学学报 ›› 2013, Vol. 33 ›› Issue (01): 30-.

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鳖甲煎丸对肝细胞癌中Wnt/β-catenin信号通路及抑制基因DKK-1、FrpHe表达的影响

贺松其,程旸,朱云,范钦,孙海涛,贾文燕   

  • 出版日期:2013-01-20 发布日期:2013-01-20

Effect of Biejiajian Pills on Wnt/β-catenin signal pathway and DKK-1 and FrpHe gene expressions in hepatocellular carcinoma cells

  • Online:2013-01-20 Published:2013-01-20

摘要: :目的通过研究鳖甲煎丸对肝细胞癌中Wnt 信号通路及抑制基因DKK-1、FrpHe 表达的影响,探讨其抗肝细胞癌转移
侵袭的作用机制与Wnt/β-catenin 信号通路的关系。方法24 只Wistar 大鼠随机分为3 组(8 只/组),分别以临床剂量的20 倍、
10 倍的鳖甲煎丸和生理盐水灌胃3 d,3 d 后采血,离心,获取血清。分别将3 组血清加入DMEM培养液中培养肝癌细胞
HepG2,48 h 后采用流式细胞术检测细胞中的β-catenin 蛋白含量,qRT-PCR法检测DKK-1、FrpHe基因的表达情况,以进一步
阐明药物的作用机制与Wnt/β-catenin 信号通路的关系。结果流式细胞术结果:高剂量组和中剂量组含药血清均可显著降
低细胞中β-catenin 蛋白表达,且该作用与药物浓度有关。RT-PCR结果:高剂量组和中剂量组含药血清均可显著下调DKK-1
基因的表达,且该作用与药物浓度有关。而高剂量组和中剂量组含药血清对FrpHe 基因的表达影响不明显。结论鳖甲煎
丸能显著抑制肝细胞癌的生长、粘附和转移,且这种抑制作用与显著降低肝癌细胞中β-catenin 蛋白表达、显著下调DKK-1 基
因的表达,从而阻断Wnt/β-catenin 信号通路有关。

Abstract: Objective To investigate the effect of Biejiajian Pills on Wnt signal pathway and its inhibitory gene (DKK-1 and
FrpHe) expressions and explore the mechanism underlying the action of Biejiajian Pills to suppress the invasiveness of
hepatocellular carcinoma. Methods Twenty-four Wistar rats were randomized equally into 3 groups for gavage of normal
saline and Biejiajian Pills at 20- and 10-fold clinical doses for 3 days. Blood samples were then collected from the rats, and the
serum was separated and added in HepG2 cell cultures. After 48 h of culture, the cells were collected to determine the cellular
content of β-catenin protein using flow cytometry and detect DKK-1 and FrpHe mRNA expressions using qRT-PCR. Results
HepG2 cells cultured in the presence of sera from rats fed with Biejiajian Pills showed significantly lowered β-catenin protein
expression and obvious down-regulation of DKK-1 mRNA expression, and the effect was correlated with the doses of the drug
administered. The expression of FrpHe mRNA showed no significant differences between the 3 groups. Conclusions Biejiajian
Pills can effectively inhibit the invasiveness and migration of hepatocellular carcinoma cells, which is closely related to
decreased expressions of β-catenin and DKK-1 to cause block of the Wnt/β-catenin signal pathway.