BMS-345541对人骨肉瘤细胞MG63凋亡及其作用机制

Apoptosis effect of BMS-345541 on human osteosarcoma MG63 cells and its mechanism

目的研究BMS-345541对人骨肉瘤细胞MG63凋亡的影响及其可能的机制。方法以0、2、4、6、8、10μmol/L BMS-345541处理MG63细胞24、48 h后,用CCK-8法检测细胞存活率。用0、2、4、8μmol/L药物作用细胞24 h后,以PI染色法观察细胞凋亡情况、流式细胞仪检测细胞凋亡。以同样浓度作用细胞48 h,Western-blot检测cleaved Caspase-3蛋白表达情况。结果 (4-10)μmol/L的BMS-345541均可抑制MG63细胞的增殖,其抑制效应呈剂量-时间依赖性,48 h最低存活率只有(0.412±0.024)(P<0.05)。PI染色荧光显微镜观察结果发现:随药物浓度增高,相同视野下正常细胞数逐渐降低,凋亡细胞数逐渐增高。流式细胞仪分析:0、2、4、8μmol/L的作用BMS-345541 24 h后,凋亡率分别为(5.2±0.78)%、(5.82±0.82)%、(8.86±1.71)%、(11.01±2.69)%,与对照组相比,8μmol/L组差异有统计学意义(P<0.05)。Western blot结果显示MG 63细胞中的cleaved caspase-3蛋白表达量上调,与对照组相比,2、4、8μmol/L组差异均有统计学意义(P<0.05)。结论 BMS-345541可抑制MG63细胞增殖,并诱导其发生凋亡,其作用机制可能与cleaved caspase-3表达上调有关。

Objective To study the effect of BMS-345541 on apoptosis of human osteosarcoma cell MG63 and its possible mechanism. Methods Treated MG63 cells with 0（blank control）, 2, 4, 6, 8, 10 μmol/L BMS-345541 for 24 h and 48 h, respctively, cell viability were detected by CCK-8 methodlThe group of 0, 2, 4, 8 μmol/L BMS-345541 effect on cells for 24 h, respectively. Apoptosis was respected by PI staining, apoptosis rate of cell was analyzed by flow cytometry. Treated cells with the same concentration for 48 h, cleaved caspase-3 protein was identified by Western-blot. Results （4- 10） μmol/L of BMS-345541 inhibited the proliferation of MG63 cells in a dose-dependent and time-dependent manner. The minimum survival rate was only （0.412±0.024）（P〈0.05）. PI staining showed that with the increase of drug concentration, the number of normal cells decreased gradually and the number of apoptotie cells increased gradually. Flow cytometry analysis show： The percentages of apoptpsis after the treatment with BMS-345541 at 0, 2, 4, 8 μmol/L for 24h were （5.2±0.78）%, （5.82±0.82） %, （8.86± 1.71 ） % and （ 11.01 ±2.69） %, respectively. The data compared with the control group, the group of 8 Ixmol/L was statistically significant （P〈0.05）. Western blot results showed that the expression of cleaved caspase-3 protein in MG 63 cells was up-regulated. The data compared with the control group, the concentration of 2, 4, 8 μmol/L was statis- tically significant （P〈0.05）. Conclusion BMS-345541 can inhibit the proliferation of MG63 cells and induce apoptosis. The mechanism may be related to the up-regulation of cleaved easpase-3 expression