【摘要】 目的探讨汉防已甲素(Tet)对人食管癌细胞株Eca-109增殖和凋亡的影响。方法将人食管癌细胞株Eca-109设不同浓度Tet处理组,并设立空白对照组。采用MTT法观察Tet对Eca-109细胞体外生长的抑制作用;采用透射电镜、流式细胞术观察Tet对Eca-109细胞凋亡的影响;采用Western blotting、RT-PCR观察Tet对Eca-109细胞凋亡相关蛋白和mRNA表达的影响。结果 Tet对Eca-109细胞的抑制作用呈时间-剂量依赖性(P<0.05),Tet干预48、72h Eca-109细胞的IC50分别为(20.886±0.215)μmol/L和(14.352±0.102)μmol/L。30μmol/L Tet处理细胞48h后,电镜下可见细胞缩小,核裂解,凋亡小体形成。分别以20、30、40μmol/L的Tet干预Eca-109细胞48h后,流式细胞术显示早期凋亡率依次为0.12%、0.34%和0.31%,中晚期凋亡率依次为4.02%、7.43%和77.42%;RT-PCR和Western blotting显示,Bcl-2蛋白和mRNA表达均随Tet浓度的增加而逐渐下调(P<0.05),Bax蛋白和mRNA表达均随Tet浓度增加而逐渐上调(P<0.05),而p53蛋白水平在处理前后无明显变化。结论 Tet可以抑制食管癌Eca-109细胞生长,促进其凋亡,其作用机制可能与下调Bcl-2表达,上调Bax表达有关。更多还原

【Abstract】 Objective To investigate the effects of tetrandrine(Tet) on human esophageal cancer cell line Eca-109 proliferation and apoptosis. Methods Human esophageal cancer cell line Eca-109 was treated with different concentrations of Tet,and blank group was established as control.Cell growth was determined by MTT,cell apoptosis was detected by transmission electron microscopy and flow cytometry,and apoptosis-related gene expression was detected by Western blotting and RT-PCR. Results The inhibiting role of Tet of on Eca-109 cells proliferation was in time-dose dependence(P<0.05).The IC50of Tet-treated 48h and 72h was(20.886±0.215)μmol/L and(14.352±0.102)μmol/L,respectively.After 48h of 30μmol/L Tet-treatment,cells showed shrink,nuclear lysis,and apoptotic body formation under electron microscope.When Eca-109 cells was treated with 20,30 and 40μmol/L Tet for 48h,cells early apoptosis rates was 0.12%,0.34% and 0.31%,and late apoptosis rates was 4.02%,7.43% and 77.42%,respectively.Furthermore,as Tet concentrations increasing,Bcl-2 protein and mRNA expression was both gradually down regulated,while Bax protein and mRNA expression was gradually up regulated(P<0.05).However,p53 protein level did not change significantly after Tet-treatment. Conclusion Tet can inhibit the growth of esophageal carcinoma Eca-109 cells and promote apoptosis in vitro.The mechanism may be related to Bcl-2 expression reduction and Bax expression increase.更多还原