六神丸和砷化合物对人肝癌Hep3B细胞增殖的影响

Effects of Liu-Shen-Wan and arsenic compounds on hepatoma Hep3B cells

目的中药六神丸(LSW)含雄黄(As_4S_4)。近年研究表明砷化合物对血液肿瘤有显著的抑制作用,但其对实体瘤的作用报道少。故我们就六神丸和砷化合物[Na_2HAsO_4(As^(5+))、NaAsO_2(As^(3+))、As_2O_3、As_4S_4和As_2S_2]对人肝癌Hep3B细胞的抗肿瘤作用进行了研究。方法采用MTS法检测六神丸、雄黄及其他含砷化合物对人肝癌Hep3B细胞的杀灭作用,通过原子吸收光谱测定各砷化合物中砷(As)的溶出度;流式细胞仪检测药物对细胞周期和细胞凋亡的影响;荧光定量PCR检测相关基因的表达。结果六神丸中As的溶出度高于雄黄和As_2S_2,但低于亚砷酸钠(As^(3+))、砷酸钠(As^(5+))和As_2O_3。六神丸对Hep3B有较好的抑制增殖和杀灭作用其作用仅次于亚砷酸钠。六神丸可诱导Hep3B细胞凋亡,作用机制与降低Bcl-2及Bcl-w的表达有关,并降低ABCG2、VEGF(血管内皮生长因子)和肿瘤转移相关基因MMP-9的表达。但六神丸对细胞周期无明显影响。结论六神丸通过多途径对肝癌Hep3B细胞具有明显的抗增殖作用。

Objective Arsenic（As） is effective for blood malignancies but little is known about its effects on solid tumors.This study aimed to examine the effect of As-containing Liu-Shen-Wan（LSW） and As compounds[Na_2HAsO_4（As^（5＋））,NaAsO_2（As^（3＋））,As_2O_3,As_4S_4 and As_2S_2]on the proliferation of the human hepatoma Hep3 B cells and the potential mechanisms.Methods Cultured Hep3 B cells were treated with LSW and arsenic compounds for 6-48 h.The cytotoxicity,cellular arsenic accumulation,cell cycle regulation,and the expression of genes related to apoptosis were examined.Results The release of As from LSW was higher than As_4S_4 and As_2S_2,but was much lower than NaAsO_2,Na_2HAsO_4,and As_2O_3.LSW was effective in inhibition of Hep3 B cell proliferation,an effect secondary to NaAsO_2,but stronger than As_4S_4.LSW induced apoptosis of the cells,but had no major effects on cell cycles.LSW down-regulated the expression of Bcl-2 and Bcl-w,and inhibited the gene expression of breast cancer resistance protein（BCRP/ABGC2）,vascular endothelial growth factor（VEGF）and matrix metallopeptidase 9（MMP-9）.Conclusion Arsenic release depends on chemical forms of arsenicals.LSW showed antitumor effects towards hepatoma Hep3 B cells,probably though apoptosis and tumor growth-related gene expressions.