吡柔比星联合二烯丙三硫抑制膀胱癌细胞生长的协同作用研究

目的探讨吡柔比星(THP)联合二烯丙三硫(DATS)对膀胱癌细胞T24的协同杀伤效应及其机制。方法 MTT法、TUNEL法、流式细胞仪分别检测THP、DATS及二者联合对T24细胞生长、细胞周期及凋亡的影响;western-blot测定Bcl-2、Bax的表达变化;比色法测定caspase-3活性。结果 THP、DATS均能抑制T24细胞生长,DATS可明显提高THP的杀伤效应;THP(0.25mg/L)联合DATS(3mg/L)作用48h的细胞生存率为(32.3±2.6)%,低于单独应用THP(63.2±1.4)%和DATS(61.9±4.2)%,二者具有协同抑瘤作用;联合药物组可明显提高细胞凋亡率及G2/M期细胞比率,并更能显著抑制Bcl-2的表达,促进caspase-3的活性增加。结论 THP联合DATS可通过共同诱导肿瘤细胞凋亡发挥协同作用,抑制膀胱癌细胞生长,该机制可能与细胞周期阻滞、Bcl-2的表达及caspase-3的活性变化有关。

The Synergetic Effect of Cell Killing by Pirarubicin Combined with Diallyl Trisulfide in Bladder Cancer Cells

Objective To investigate the killing effect of pirarubicin (THP) combined with diallyl trisulfide (DATS) on bladder cancer cell line T24 and further to determine the mechanism. Methods T24 cells were treated with various concentrations of THP or (and) DATS. Cell viability was estimated using MTT assay. Apoptotic cells were detected using TUNEL assay. The percentage of sub G1 and cell cycle were confirmed by flow cytometry. Bcl-2 and Bax protein levels were determined using western bolt. The activity of caspase-3 was measured using a colorimetric assay. Results THP or DATS alone could all inhibit the proliferation of T24 cells. For combination group, the cell viability was decreased to(32.3 ± 2.6)% to compare with(63.2 ± 1.4)% of THP at 0.25 mg/L and(61.9 ± 4.2)% of DATS at 3 mg/L separately and there was a synergistic effect between these two drugs. Apoptotic rate and the cell number in G2/M-phase of the combination group were higher than other two drugs. The combination treatment could significantly reduce the Bc1-2 expression and enhance the activity of caspase-3. Conclusions The combination of THP with DATS can inhibit the proliferation of bladder cancer cells congenerously through inducing apoptosis, which may be correlated with G2/M-phase arrest, the down-regulation of Bcl-2 and the activation of caspase-3.