联合应用三氧化二砷与紫杉醇诱导小鼠结肠癌CT26细胞凋亡及其机制研究

Combination treatment of arsenic trioxide and paclitaxel to induce apoptosis in mouse colon cancer CT26 cells and its mechanism

目的探讨联合应用三氧化二砷(arsenic trioxide,As2O3)与紫杉醇(paclitaxel,PTX)诱导小鼠结肠癌CT26细胞凋亡的作用及其机制。方法分别以不同浓度的As2O3CT26细胞72 h,以MTT法检测细胞增殖率,选择30%左右抑制浓度的As2O3与不同浓度的紫杉醇联合处理细胞,MTT法检测细胞增值率,以抑制增殖效果最为明显的剂量组进行后续实验;以流式细胞仪技术检测药物处理后各组细胞周期及凋亡率,采用Western blot法检测药物处理后,细胞中Bax,Blc-2以及PARP的表达情况。结果不同浓度的As2O3和不同浓度的PTX处理CT26细胞后,均能够显著抑制细胞增殖,呈剂量效应关系;2μmol/L As2O3与0.04μmol/L PTX联合处理能够协同抑制CT26细胞增殖;与As2O3以及PTX单独处理组相比,两者联合作用能够显著增加细胞凋亡率,上调细胞中Bax/Bcl-2表达以及使PARP活化(均P<0.05)。结论 As2O3与PTX能够协同抑制结肠癌CT26细胞增殖、诱导细胞凋亡,其作用机制可能与上调细胞中Bax/Bcl-2和活化PARP蛋白有关。

Objective To study the effect of arsenic trioxide （ As2O3 ） and paclitaxel （PTX） to induce apoptosis in mouse colon cancer CT26 cells and its mechanism. Methods Cells were treated with different concentrations of As2O3, and cell growth rate was detected by MTY assay, Then cells were treated with As2O3 at 30% effective close cooperative with different concentrations of PTX to find out tle most effective Combination dose on cell grouwth inhibition. Apoptosis rate and cell cycle of each group were assessed by flow cytometry. Western blot analysis was used to detect the protein expression of Bax, Bcl-2 and PARP of cells. Results As203 or PTX alone inhibited CT26 cell growth in a dose dependent manner. Combination treatment of As2O3 at 2 μmol/L and PTX at 0. 04μmol/L showed the best effect in cell growth inhibition, as well as induced increasing level of apoptosis rate of cells compared with single treatment. Combination treatment of As2O3 and PTX significantly up-regulated apoptosis rate and Bax and PARP cleavage, down-regulated Bcl-2 expression compared with As2O3 and PTX alone （ all P 〈 0. 05 ）. Conclusion Combination treatment of As2O3 and PTX inhibit cell growth and induce apoptosis through up-regulating Bax/Bel-2 and PARP cleavage.