The noncoding RNA expression profile and the effect of lncRNA AK126698 on cisplatin resistance in non-small-cell lung cancer cell

PLoS One. 2013 May 31;8(5):e65309. doi: 10.1371/journal.pone.0065309. Print 2013.

Abstract

Background: The efficacy of cisplatin-based chemotherapy in non-small-cell lung cancer is limited by the acquired drug resistance. Identification the RNAs related to the cisplatin resistance may help to improve clinical response rates.

Methods: Microarray expression profiling of mRNAs, lncRNA and miRNA was undertaken in A549 cells and cisplatin resistant A549/CDDP cells. Differentially expressed mRNAs, lncRNAs and miRNAs, verified by realtime RT-PCR, were subjected to pathway analysis. Expression of NKD2 and β-catenin was assessed by realtime RT-PCR and western blot analysis. The effect of lncRNA AK126698 on cisplatin induced apoptosis was investigated by annexin-V/PI flow cytometry.

Results: In total, 1471 mRNAs, 1380 lncRNAs and 25 miRNAs differentially expressed in A549/CDDP and A549 cells. Among them, 8 mRNAs, 8 lncRNAs and 5 miRNAs differentially expressed in gene chip analysis were validated. High-enrichment pathway analysis identified that some classical pathways participated in proliferation, differentiation, avoidance of apoptosis, and drug metabolism were differently expressed in these cells lines. Gene co-expression network identified many genes like FN1, CTSB, EGFR, and NKD2; lncRNAs including BX648420, ENST00000366408, and AK126698; and miRNAs such as miR-26a and let-7i potentially played a key role in cisplatin resistance. Among which, the canonical Wnt pathway was investigated because it was demonstrated to be targeted by both lncRNAs and miRNAs including lncRNA AK126698. Knockdown lncRNA AK126698 not only greatly decreased NKD2 which can negatively regulate Wnt/β-catenin signaling but also increased the accumulation and nuclear translocation of β-catenin, and significantly depressed apoptosis rate induced by cisplatin in A549 cells.

Conclusion: Cisplatin resistance in non-small-cell lung cancer cells may relate to the changes in noncoding RNAs. Among these, AK126698 appears to confer cisplatin resistance by targeting the Wnt pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carcinoma, Non-Small-Cell Lung / genetics*
  • Carcinoma, Non-Small-Cell Lung / metabolism
  • Cell Line, Tumor
  • Cisplatin / pharmacology
  • Cluster Analysis
  • Drug Resistance, Neoplasm / genetics*
  • Gene Expression Profiling
  • Gene Knockdown Techniques
  • Gene Regulatory Networks
  • Humans
  • Lung Neoplasms / genetics*
  • Lung Neoplasms / metabolism
  • RNA, Long Noncoding / genetics*
  • RNA, Untranslated / genetics*
  • Reproducibility of Results
  • Signal Transduction
  • Transcriptome*
  • Wnt Signaling Pathway

Substances

  • RNA, Long Noncoding
  • RNA, Untranslated
  • Cisplatin

Grants and funding

The work was supported by the Natural Science Foundation of China (No. 81071910, 81070042). LncRNA microarray experiments were performed by KangChen Bio-tech, Shanghai, China. MiRNA microarray experiments were performed by Genminix Informatics Ltd., Shanghai, China. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.