摘要
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 determine the effect of tubeimoside 1(TBMS1) in enhancing chemosensitivity of drug-resistant ovarian cancer cell line(A2780/DDP) to cisplatin.MethodsThe A2780/DDP cells were treated with different concentrations of cisplatin and TBMS1 to analyze 50% inhibiting concentration(IC50) by methyl thiazolyl tetrazolium(MTT) assay.Experiment was divided into 4 groups.Control group:treatment with physiological saline;cisplatin group:treatment with 6 μmol/L cisplatin;Combination of drugs groupⅠ:6 μmol/L cisplatin + 3 μmol/L TBMS1;Combination of drugs groupⅡ:6 μmol/L cisplatin + 6 μmol/L TBMS1.After the cells were treated by the above methods,the cells viability was analyzed by MTT assay.The cellular DNA damage was measured by agarose gel electrophoresis.Apoptosis and cell cycles were measured by flow cytometry.The expression of Cyclin A and Cyclin D1 protein was determined by Western blotting.Results TBMS1(3 and 6 μmol/L) combined with cisplatin(6 μmol/L) enhanced the chemosensitivity of A2780/DDP cells to cisplatin.After co-treatment with TBMS1,the level of cellular DNA damage was significantly increased with increase of TBMS1 concentration.Cell cycle was arrested in S phase,with a proportion of(51.23±3.53) % and(77.34±2.80)%,respectively for groupⅠ and group Ⅱ.Apoptosis ratio was(5.19±0.78)%,and(7.11±1.06)% respectively,Cyclin A expression of was 1.89±0.28 and 2.91±0.43 respectively,while that of Cyclin D1 was 0.66±0.09 and 0.28±0.05 respectively for the 2 groups(P0.01,0.05).Conclusion TBMS1 inhibits the growth of A2780/DDP cells and enhances the sensitivity to cisplatin,and Cyclin A and Cyclin D1 might be involved in the processes.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2011年第7期695-698,共4页
Journal of Third Military Medical University
基金
重庆市自然科学基金(CSTC2009BA5083)
重庆医科大学重点基金(XBZD200806)~~