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Tumor Biology

, Volume 37, Issue 2, pp 2387–2394 | Cite as

Inhibition of microRNA-196a might reverse cisplatin resistance of A549/DDP non-small-cell lung cancer cell line

  • Jian-Huang Li
  • Ning Luo
  • Mei-Zuo Zhong
  • Zhi-Qiang Xiao
  • Jian-Xin Wang
  • Xiao-Yi Yao
  • Yun Peng
  • Jun Cao
Original Article

Abstract

We aimed to explore the possible mechanism of microRNA-196a (miR-196a) inhibition and reversion of drug resistance to cisplatin (DDP) of the A549/DDP non-small-cell lung cancer (NSCLC) cell line. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect expression differences of miR-196a in the drug-resistant A549/DDP NLCLC cell line and the parental A549 cell line, and expressions of miR-196a in the A549/DDP NLCLC cell line transfected with miR-196a inhibitor (anti-miR-196a group) and the miR-196a negative control (miR-NC) group and blank group (without transfection). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test was applied in examining the cell viability of A549/DDP cell line before and after transfection. Clonogenic assay was used to detect cell proliferation ability. Flow cytometry was applied in detecting apoptosis rate of assayed tumor cell and rhodamine-123 changes in cells. Western blot was applied in detecting proteins of drug-resistant related gene in A549/DDP cell line. Significantly higher expression of miR-196a was detected in the drug-resistant A549/DDP cell line than that in the parental A549 cell line (P < 0.05). However, miR-196a expression in the anti-miR-196a group decreased obviously compared to that in the blank group and the miR-NC group (both P < 0.05); The value of IC50 in the anti-miR-196a group showed remarkably lower than that in the blank group and the miR-NC group (both P < 0.05); Rh-123 absorbing ability in the anti-miR-196a group increased 2.51 times and 2.49 times respectively compared to that in the blank group and the miR-NC group (both P < 0.05). No statistical differences in the apoptosis rate of A549/DDP cell line in the early stage were found among the three groups (all P > 0.05), but the late-stage apoptosis rate in the anti-miR-196a group was significantly higher than that in the blank group and the miR-NC group (both P < 0.05); The expressions of human multidrug resistance 1 (MDR1), multidrug resistance protein 1 (MRP1), excision repair cross-complementation 1 (ERCC1), survivin, and B cell lymphoma 2 (Bcl-2) decreased significantly while RhoE increased significantly in the anti-miR-196a group than the blank group and the miR-NC group (all P < 0.05). Inhibition of miR-196a could reverse cisplatin resistance of A549/DDP cell lines, which might relate with inhibition of drug efflux, down-regulation of drug-resistant protein expression, cell apoptosis, and cell proliferation suppression.

Keywords

MicroRNA-196a Cisplatin resistance A549/DDP cell lines NSCLC 

Notes

Acknowledgments

This study was funded by the Key Program of the National Natural Science Foundation of China (No. 61232001). We would like to acknowledge the reviewers for their helpful comments on this paper.

Authors’ contributions

JH Li and N Luo designed the study. MZ Zhong and ZQ Xiao conceived and supervised the study. JX Wang performed the examination and the analysis. XY Yao performed the statistical analysis. Y Peng and J Cao interpreted the results. JH Li and J Cao drafted and revised the paper. All authors read and approved the final paper.

Conflicts of interest

None

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Jian-Huang Li
    • 1
  • Ning Luo
    • 1
  • Mei-Zuo Zhong
    • 1
  • Zhi-Qiang Xiao
    • 2
  • Jian-Xin Wang
    • 3
  • Xiao-Yi Yao
    • 1
  • Yun Peng
    • 1
  • Jun Cao
    • 4
  1. 1.Department of Oncology, Xiangya HospitalCentral South UniversityChangshaPeople’s Republic of China
  2. 2.Research Center of Carcinogenesis and Targeted Therapy, Xiangya HospitalCentral South UniversityChangshaPeople’s Republic of China
  3. 3.School of Information Science and EngineeringCentral South UniversityChangshaPeople’s Republic of China
  4. 4.Department of Medical Oncology, the Affiliated Cancer Hospital of Xiangya School of MedicineCentral South UniversityChangshaPeople’s Republic of China

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