The Reversing and Molecular Mechanisms of miR-503 on the Drug-resistance to Cisplatin in A549/DDP Cells
10.3779/j.issn.1009-3419.2014.01.01
- VernacularTitle:MiR-503逆转肺癌耐药细胞株A549/DDP的耐药性及其机制研究
- Author:
WU YI
1
;
GUO LILI
;
LIU JINGHAO
;
LIU RENWANG
;
LIU MINGHUI
;
CHEN JUN
Author Information
1. 300052天津，天津医科大学总医院肺部肿瘤外科，天津市肺癌研究所，天津市肺癌转移与肿瘤微环境实验室
- Keywords:
MicroRNA;
Lung neoplasms;
Drug-resistant
- From:
Chinese Journal of Lung Cancer
2014;(1):1-7
- CountryChina
- Language:Chinese
-
Abstract:
Background and objective Cisplatin-resistance in lung cancer cells is general in clinic, hence it is sig-niifcant to investigate the mechanisms of cisplatin-resistant and develop new methods of reversing drug-resistance. Recent researches showed that miRNA could regulate cell growth, apoptosis, migration and invasion even in drug therapy in cancer by its target gene. hTe aim of this study is to investigate the effects and molecular mechanisms of miR-503 on reversing the cisplatin-resistance in lung cancer DDP-resistant cell line A549/DDP. Methods MTS assay was employed to determine the effect of miR-503 on A549/DDP’ sensitivity to cisplatin. Apoptosis rate and intracellular concentration of rhodamine-123 (Rh-123) were determined by lfow cytometry, the expression of multi-drugs resistant proteins MDR1and MRP1, ERCC1, RhoE, Survivin and Bcl-2 were determined by Western blot and real time PCR. hTe phosphorylation of Akt was analyzed by Western blot, the transcriptional activities of NF-κB and AP-1were detected by dual-luciferase reporter gene systems. Results MiR-503 was able to increase the cisplatin sensitivity of A549/DDP. Atfer treatment with miR-503, the reverse folds (RF) to cisplatin was 2.48 fold, the intracellular level of Rh-123 was 2.49 fold, the apoptosis rate was10.3 fold, the expressions of several drug-resistant related proteins, such as MDR1, MRP1, ERCC1, Survivin and Bcl-2 were downregulated signiifcantly, as shown by WB, in contrast, the level of RhoE was elevated, the mRNA epression of MDR1was18.5%, the mRNA epression of MRP1was 22.3%, the mRNA epression of ERCC1was18.6%, the mRNA epression of Survivin was 42.8%, the mRNA expression of Bcl-2 was 68.1%, the mRNA epression of RhoE was 206.5%, in addition, the phosphorylation of Akt decreased and transcrip-tional activities of NF-κB was 53.7%, AP-1was 47.4%compared with control group. Conclusion MiR-503 was able to reverse the cisplatin resistance of A549/DDP. MiR-503 processed this kind of effect by inhibiting the drug effux, downregulating the expression of drug-resistant related proteins and promoting cell apoptosis.
