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目的探讨土贝母皂苷甲(tubeimoside 1,TBMS1)促进人卵巢癌A2780/DDP细胞对顺铂(cisplatin,DDP)的增敏作用。方法采用MTT法分别检测顺铂和TBMS1对A2780/DDP细胞作用的半抑制浓度值(IC50),将实验分为对照组:只加等量生理盐水;顺铂组:给予6μmol/L顺铂处理;联合用药Ⅰ组:6μmol/L顺铂+3μmol/L TBMS1;联合用药Ⅱ组:6μmol/L顺铂+6μmol/L TBMS1。按上述方法处理细胞后,MTT法检测细胞活力;电泳法检测细胞内DNA损伤;流式细胞技术检测细胞凋亡及周期;Western blot分析周期蛋白A(Cyclin A)和周期蛋白D1(Cyclin D1)的表达。结果 TBMS1(3、6μmol/L)与顺铂(6μmol/L)联用使人卵巢癌A2780/DDP细胞对顺铂的敏感性增强;细胞的DNA损伤随TBMS1浓度的增加而明显增多;细胞周期被阻滞在S期,联合用药Ⅰ和Ⅱ组S期所占比例分别为(51.23±3.53)%、(77.34±2.80)%;细胞凋亡率为(5.19±0.78)%、(7.11±1.06)%;Cyclin A的表达为1.89±0.28及2.91±0.43,CyclinD1为0.66±0.09及0.28±0.05(P<0.05,P<0.01)。结论 TBMS1显著增加顺铂对A2780/DDP细胞增殖抑制作用,Cyclin A、Cyclin D1可能参与其调节。
Objective To investigate the effect of tubeimoside 1 (TBMS1) on the sensitization of human ovarian cancer A2780 / DDP cells to cisplatin (DDP). Methods The half inhibitory concentration (IC50) of cisplatin and the effect of TBMS1 on A2780 / DDP cells were determined by MTT method. The experiment was divided into control group: only equal amount of saline; cisplatin group: 6μmol / L cisplatin ; Combination group Ⅰ: 6μmol / L cisplatin + 3μmol / L TBMS1; combination group Ⅱ: 6μmol / L cisplatin + 6μmol / L TBMS1. Cell viability was detected by MTT assay, DNA damage in cells was detected by electrophoresis, apoptosis and cell cycle were detected by flow cytometry, Western blot analysis was used to detect Cyclin A and Cyclin D1, expression. Results The combination of TBMS1 (3,6μmol / L) and cisplatin (6μmol / L) enhanced the sensitivity of human ovarian cancer A2780 / DDP cells to cisplatin. The DNA damage of cells increased with the increase of TBMS1 concentration. (51.23 ± 3.53)% and (77.34 ± 2.80)%, respectively. The rate of apoptosis was (5.19 ± 0.78)%, (7.11 ± 1.06) )%; Cyclin A expression was 1.89 ± 0.28 and 2.91 ± 0.43, CyclinD1 was 0.66 ± 0.09 and 0.28 ± 0.05 (P <0.05, P <0.01). Conclusion TBMS1 significantly increased cisplatin-induced proliferation of A2780 / DDP cells, and Cyclin A and Cyclin D1 may be involved in the regulation.