Objective: To study the effect of PX-12 on apoptosis of multiple myeloma (MM) cell line induced by bortezomib.
Methods: MM cell line H929 cells were divided into PX-12 group, bortezomib group, combination group, and control group. 5.0 μmol/L PX-12, 20 nmol/L bortezomib, combination of the two drugs, and DMSO were given to the above mentioned group, respectively. After culture for 24, 48, and 72 hours, the changes of cell viability were observed, the MM cell activity was detected by MTT method, and the cell cycle distribution and apoptosis of each group was detected by flow cytometry. The intracellular ROS level was measured by H2DCFDA probe labeling.
Results: MTT assay showed that after culture for 72 hours, the activity of H929 cells in PX-12 group (P<0.05) and bortezomib group (P<0.01) was significantly lower than that in the control group, while that in the combination group was decreased most significantly (P<0.01). After culture for 48 hours, cells in G1 phase in PX-12 group was decreased to 40%, while cells in S phase and G2/M phase was increased to 28% and 40%, respectively. The cells in bortezomib group also showed a similar distribution after being treated. After treated with PX-12 and bortezomib, the cells in G1 phase were decreased significantly to 19% and 12% in S phase, but increased significantly to 68% in G2/M phase, which was significantly different from PX-12 group and bortezomib group (P<0.01). After culture for 72 hours, the apoptosis rate was 71.3% in the combination group, which was significantly higher than that in PX-12 group, bortezomib group, and control group (20.6%, 33.3%, 10.6%)(P<0.01). After culture for 24 hours, the intracellular ROS level in the combination group was 12015±430.2, which was higher than that in the PX-12 group, bortezomib group, and control group (6729±352.8, 2651±228.3, 1098±164.6, respectively) (P<0.01).
Conclusion: PX-12 can increase the apoptosis of MM cell line H929 induced by bortezomib, which may be caused by increasing of ROS level.
题目: PX-12促进硼替佐米诱导多发性骨髓瘤细胞H929凋亡的实验研究.
目的: 研究PX-12对硼替佐米诱导多发性骨髓瘤（MM）细胞凋亡的影响及机制.
方法: 以MM细胞株H929细胞为研究对象，将细胞分为PX-12组、硼替佐米组、联合组和对照组，分别加入单药PX-12 5.0 μmol/L、单药硼替佐米20 nmol/L、两药联合和DMSO对照，培养24、48及72 h后观察细胞活性变化，通过MTT法检测MM细胞活性，采用流式细胞术检测各组细胞周期分布和凋亡情况，H2DCFDA探针标记法测定各组细胞内ROS水平.
结果: 培养72 h后，PX-12组和硼替佐米组的H929细胞活性均明显低于对照组（P<0.05，P<0.01），而联合组的细胞活性下降最为显著，且明显低于其他3组（P<0.01）。PX-12组细胞在培养48 h后，G1期的细胞数下降至40%，而S期和G2/M期数目分别上升至28%和40%，硼替佐米组细胞作用后也出现类似的分布现象，而联合组细胞经过PX-12和硼替佐米共同作用后，G1期的细胞明显下降至19%，S期细胞也降至12%，但G2/M期细胞明显上升至68%，与PX-12组和硼替佐米组相比，差异有统计学意义（P<0.01）。培养72 h后，联合组细胞凋亡率达713%，显著高于PX-12组、硼替佐米组和对照组（20.6%、33.3%和10.6%）（P<0.01）。培养24 h后，联合组细胞内ROS水平为12015±430.2，分别高于PX-12组、硼替佐米组和对照组（6729±352.8、2651±228.3和1098±164.6）（P<0.01）.
结论: PX-12可以促使硼替佐米诱导MM细胞H929凋亡增加，可能是通过ROS水平升高而导致凋亡增加.