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. 2014 Jul 10:9:143.
doi: 10.1186/1746-1596-9-143.

MicroRNA-182 modulates chemosensitivity of human non-small cell lung cancer to cisplatin by targeting PDCD4

Fang-ling Ning, Feng Wang, Mian-li Li, Ze-Shun Yu, Yan-zhang Hao, Shao-shui Chen  1
Affiliations

MicroRNA-182 modulates chemosensitivity of human non-small cell lung cancer to cisplatin by targeting PDCD4

Fang-ling Ning et al. Diagn Pathol. .

Abstract

Background: Overexpression of microRNA-182 (miR-182) is found in various human cancers, including non-small cell lung cancer (NSCLC). Our aim is to investigate the association of miR-182 expression with the sensitivity of NSCLC to cisplatin.

Methods: TaqMan RT-PCR or Western blot assay was performed to detect the expression of mature miR-182 and programmed cell death 4 (PDCD4) protein. miR-182 and (or) PDCD4 depleted cell lines were generated using miR-182 inhibitor and (or) siRNA. The viabilities of treated cells were analyzed using MTT assay.

Results: The expression level of miR-182 in A549 cell line was significantly higher than that in NHBE cell line (p < 0.01). Transfection of miR-182 inhibitor induced sensitivity of A549 cells to cisplatin. A549 cells transfected with PDCD4 siRNA became more resistant to cisplatin therapy. We found an increase PDCD4 protein level following the transfection of miR-182 inhibitor using Western blot analysis. In addition, the enhanced growth-inhibitory effect by miR-182 inhibitor was weakened after the addition of PDCD4 siRNA.

Conclusions: The results of the present study demonstrated that overexpression of miR-182 may involve in chemoresistance of NSCLC cells to cisplatin by down-regulating PDCD4.

Virtual slides: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1793467320130186.

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Figures

Figure 1
Figure 1
MiR-182 was up-regulated in A549 cell line compared to that in NHBE cell line (p < 0.01).
Figure 2
Figure 2
Transfection of miR-182 inhibitor and its negative control oligonucleotides (NC) into A549 cells. qRT-PCR showed significant under-expression of miR-182 in the transfected cells compared with control cells.
Figure 3
Figure 3
MTT assay revealed that the anti-tumour effects cisplatin in the miR-182 downregulated cells were significantly profound than in control cells (*P <0.05). Data are mean ± SD of three experiments.
Figure 4
Figure 4
Evaluation of PDCD4 in A549cells transfected with miR-182 inhibitor and its negative control oligonucleotides (NC). A. qRT-PCR showed significant upregulation of PDCD4 mRNAs in the transfected cells. B. Western blot analysis demonstrated significant overexpression of PDCD4 proteins in the transfected cells (*P < 0.05). Data are mean ± SD of three experiments.
Figure 5
Figure 5
Changes in anti-tumour effects of the cisplatin after transfection of anti-miR-182 and/or siRNA against PDCD4 in A549 cells. The MTT assay indicated a weaker anti-tumour effect of cisplatin following transfection of PDCD4 siRNA, and the enhanced growth-inhibitory effect by anti-miR-182 transfection was also weakened after the addition of PDCD4 siRNA (*P <0.05).
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