狼毒大戟对小鼠Lewis肺癌细胞凋亡影响及其机制的初步探讨

Apoptosis in Lewis carcinoma cell induced by euphorbia fischeriana steud and its mechanism

目的:观察中药狼毒大戟提取液对体外培养的Lewis肺癌细胞凋亡及Caspase-9基因表达的影响。方法:Lewis细胞培养后以不同药物浓度(0.2、0.4和0.8mg/mL)干预24h。Annexin-ⅤFITC/PI双染法检测细胞凋亡率的变化,流式细胞术检测细胞周期的变化;Real-time PCR检测基因Caspase-9表达的变化。结果:阴性对照组与狼毒大戟低、中、高剂量组(0.2、0.4和0.8mg/mL)细胞凋亡率分别为(0.331±0.012)%、(8.27±0.067)%、(28.19±0.270)%和(32.96±0.14)%;阴性对照组与狼毒大戟低中高剂量组S期所占比例分别为(50.3±0.77)%、(44.2±1.82)%、(34.1±1.56)%和(25±0.72)%;Real-time PCR结果显示,阴性对照组与狼毒大戟组Caspase-9mRNA的相对表达量分别为1.00和1.64。结论:狼毒大戟可以明显促进Lewis肺癌细胞的凋亡,将细胞周期阻滞在G0/G1期;并且可以明显提高Caspase-9的表达。狼毒大戟可能是通过将细胞周期阻滞于G0/G1期并且促进Caspase-9mRNA的表达而诱导Lewis细胞的凋亡。

OBJECTIVE： To investigate the effect of euphorbia fischeriana steud（LDE）on apoptosis and expression of Caspase-9 mRNA in Lewis lung carcinoma. METHODS： Lewis cells were exposed with LDE of different concentration （0.2,0.4 and 0.8 mg/mL） for 24 hours. Annexin V/PI double label staining was applied to detect the change of apopto sis rate. Flow cytometry was applied to detect the change of cell cycle. The mRNA level of Caspase 9 was detected by Real-time PCR. RESULTS： By Annexin-V/PI double label staining to examine apoptosis of control and low,middle and high LDE dose group （0. 2,0. 4 and 0. 8 mg/mL）,and the rate respectively was （0.331±0.012）%, （8.27±0.067）%, （28. 19±0. 270）%,（32.96±0.14）%. Flow cytometryshowed that the G0/G1 phase rate of the control and low,middle and high LDE dose group were （50.3±0.77）%,（44.2±1.82）%,（34.1±1.56）%,（25±0.72）%. Real time PCR showed that Caspase-9 mRNA expression level of the control and LDE group were 1.00 and 1.64. CONCLUSION： LDE can effectively induce apoptosis of Lewis cells through blocking the cell cycle at G0/G1 phase and increase the expression of Caspase 9 mRNA level.