| 摘要: |
| 目的探讨汉防己甲素（Tet）联合阿霉素（DOX）体外逆转耐药型前列腺癌（DU145/DOX）细胞耐药的可行性及潜在机制。方法采用经典的DOX浓度梯度诱导法制备DU145/DOX细胞，然后采用3-（4，5-二甲基噻唑-2）-2，5-二苯基四氮唑溴盐（MTT）法测定Tet干预对DU145/DOX细胞生长抑制的作用，凋亡试剂盒测定联合用药对耐药细胞凋亡诱导的影响，流式细胞仪和荧光分光光度计分别测定Tet对DOX和荧光探针罗丹明123（R123）被DU145/DOX细胞摄取行为，Pgp-GloTM分析系统测定Tet对P-糖蛋白（P-gp）ATP酶活性的影响，Westernblot法测定Tet对耐药细胞膜上P-gp蛋白表达的影响，RT-PCR法测定Tet对DU145/DOX细胞MDR1mRNA表达的影响。结果Tet组、DOX组、DOX+Tet（1/5）组对DU145/DOX细胞的半数抑制浓度（IC50，以COX浓度计算）分别为（7.16±0.54）、（1.62±0.09）、（0.37±0.03）滋M，Tet能明显增强DOX的抗肿瘤作用。DOX+Tet（1/5）组诱导DU145/DOX细胞凋亡率是DOX组的2.1倍。Tet的参与能明显提高DU145/DOX细胞内DOX和R123的累积量，其机制可能与Tet刺激细胞内P-gpATP酶活性、降低细胞膜上P-gp蛋白表达有关。Tet可能通过降低耐药基因表达的机制，联合DOX提高对DU145/DOX细胞杀伤作用。结论Tet与DOX体外联合使用具有逆转DU145/DOX细胞耐药的潜力，作用机制可能涉及刺激耐药泵酶活性、降低耐药蛋白表达量、下调MDR1mRNA表达等。 |
| 关键词: 汉防己甲素 阿霉素 DU145 细胞 逆转耐药 机制 |
| DOI：10.12056/j.issn.1006-2785.2018.40.13.2017-2709 |
| 分类号: |
| 基金项目: |
|
| Combination of tetrandrine with doxorubicin reverses drug resistance of prostate carcinoma DU145/DOX cells and its molecular mechanisms |
|
|
|
Department of Anesthesiology，shulan（hangzhou） Hospital
|
| Abstract: |
| Objective To investigate the effect of combination of tetrandrine (Tet) with doxorubicin (DOX) on drug resistance of prostate carcinoma DU145/DOX cells and its molecular mechanisms. Methods Drug resistant prostate cancer DU145/DOX cells were induced, and treated with Tet, DOX or Tet+DOX, respectively. The cell growth was assayed by MTT method, cells apoptosis was determined with AnnexinV-PE/7-AAD kit, the intracellular accumulation of DOX and R123 was tested by flow cytometry and fluorospectrophotometer, the P-gp ATPase activity and P-gp expression were detected by Pgp-GloTM Assay Systems and Western blot method, respectively, the expression of MDR1 mRNA was detected by RT-PCR.
Results The IC50 of Tet, DOX and Tet+DOX against DU145/DOX cells were(7.16±0.54), (1.62 ±0.09) and (0.37 ±0.03)滋M,
respectively. The apoptosis induction ratio of DU145/DOX cells treated with Tet+DOX was 2.1-folded of DOX group. The intracellular accumulation of DOX and R123 increased significantly in the presence of Tet. The P-gp ATPase activity was elevated, the expression P-gp and MDR1 mRNA was down-regulated in DU145/DOX cells after Tet treatment. Conclusion The combination of Tet with DOX can reverse drug resistance of DU145/DOX cells in vitro, which is associated with the effects of Tet in interfering P-gp function and suppressing MDR1 mRNA expression. |
| Key words: Tetrandrine Doxorubicin DU145 cells Resistance reversal Mechanism |
