三氧化二砷和细胞分化剂诱导肝癌细胞凋亡的阈值

Apoptotic threshold of hepatoma cell lines induced by As_2O_3 and CDA- II

目的探讨三氧化二砷(As2O3)诱导肝癌细胞凋亡的阈值以及与细胞分化剂(CDA-II)的协同效应。方法1.0～5.0μmol/L的As2O3和1.0～5.0 g/L的CDA-Ⅱ分别以及联合与肝癌细胞株Bel-7402、HepG2共孵育一定时间后,用活细胞荧光染色法、DNA电泳和流式细胞术分析细胞凋亡率。结果单用As2O3 1.0μmol/L与对照组比较,差异无显著性(P>0.05),凋亡阈值是5.0μmol/L;单用CDA-Ⅱ3.0 g/L以上与对照组比较,差异有显著性(P<0.05)。但两药联合应用的凋亡阈值是1.0μmol/L As2O3+1.0 g/L CDA-II。结论 CDA-Ⅱ可以降低肝癌细胞的凋亡阈值,增加肝癌细胞对As2O3的敏感性。

Objective To study the apoptosis threshold of hepatoma cell lines induced by arsenic trioxide (As2O3) and cell differentiation agent- II (CDA- II ). Methods Hepatoma cell lines were cultured with 1.0-5.0 μmol/L As2O3 and 1. 0-5. 0 g/L CDA- II for various durations. Using immunofluo-rescence staining of Hoechst 33 258 in hepatoma cells, flow cytometry and DNA gel eletrophoresis were applied to detect the apoptosis rate. Results After treatmeant with As2O3 or CDA-II + As2O3, there were dispersive fluorescences in normal cells' nuclei and compact paniculate fluorescences in apoptosis cells' nuclei by immunofluorescence staiming of Hoechst 33 258; agarose electrophoresis showed marked DNA ladder; flow cytometry analysis revealed a sub-G1 cell peak. The apoptotic threshold of hepatoma cells was 5.0 @mol/L As2O3 or 1.0 μmol/L As2O3 + 1.0 g/L CDA- II . Conclusion CDA- II and As2O3 can induce apoptosis of the hepatoma cells, and display a significant synergic effect.