Figures
Figure 1
Identification of miR-139 as a novel p53-responsive gene.
Validation of induction of p53 and its targets gene. HCT116 p53+/+ and HCT116 p53-/- cells were treated with DMSO or 4uM INZ-C for 18 hr in duplicate and the protein samples and RNA samples were …
Figure 2
miR-139 is a direct target of p53.
(A) Diagram shows putative p53 responsive element (p53RE) located upstream of miR-139 gene. (B) Increased binding of p53 and the endogenous p53RE-containing miR-139 promoter in response to …
Figure 3 with 3 supplements
p53 modulates PDE4D expression via miR-139-5p.
(A) Bioinformatic analysis shows the miR-139-5p-targeted 3’-untranslated region (3’UTR) sequence of PDE4D mRNA. (B) Overexpression of miR-139-5p decreases the level of PDE4D protein in cells. H460 …
Figure 3—figure supplement 1
HepG2 cells were collected 48 hr after transfection with miR-139-5p mimic at 0, 20 or 40 nM for Western blot analysis on IGF-IR expression.
https://doi.org/10.7554/eLife.15978.006
Figure 3—figure supplement 2
PC-3 cells were collected 48 hr after transfection with miR-139-5p mimic at 40 nM for Western blot analysis on PDE4D expression.
https://doi.org/10.7554/eLife.15978.007
Figure 3—figure supplement 3
HCT116 p53+/+ and HCT116 p53-/- cells were treated with 10 μM Nutlin-3 (Nut) or 5 nM Actinomycin D (ActD) for 18 hr, and then collected for Western blot analysis.
https://doi.org/10.7554/eLife.15978.008
Figure 4 with 6 supplements
miR-139-5p induces cAMP/BIM mediated cell growth arrest.
(A). Overexpression of miR-139-5p increases cellular cAMP levels. A549 cells were transfected with miR-139-5p mimic control or miR-139-5p mimic and were subjected to cAMP measurement 48 hr after …
Figure 4—figure supplement 1
Doxorubicin increases cellular cAMP levels via miR-139-5p.
A549 cells were transfected with miR-139-5p inhibitor control or miR-139-5p inhibitor for 48 hr followed by 0.5 μM Doxorubicin treatment for 18 hr and subjected to cAMP measurement.
Figure 4—figure supplement 2
H1299 cells were treated with vehicle or 0.5 μM Doxorubicin for 18 hr followed by cAMP measurement, or transfected with miR-139-5p mimic control or miR-139-5p mimic and subjected to cAMP measurement 48 hr after transfection.
https://doi.org/10.7554/eLife.15978.011
Figure 4—figure supplement 3
miR-139-5p inhibitor rescues Nutlin-3 inhibition of cell growth.
A549 cells were transfected with miR-139-5p inhibitor control or miR-139-5p inhibitor for 48 hr followed by 10 μM Nutlin-3 treatment for 24 hr and subjected to MTT analysis. *p<0.05 as compared to …
Figure 4—figure supplement 4
HCT116 p53+/+ and HCT116 p53-/- cells were treated with vehicle, or 10 μM Nutlin-3 for 18 hr, and subjected to Western blot analysis.
https://doi.org/10.7554/eLife.15978.013
Figure 4—figure supplement 5
BIM knockdown alleviates Nutlin-3 inhibition of cell growth.
A549 cells were transfected with control siRNA or BIM siRNA and 48 hr after transfection, the cells were treated with 10 μM Nutlin-3 for 24 hr, followed by MTT assay and Western blot analysis. Error …
Figure 4—figure supplement 6
A549 cells were transfected with miRNA inhibitor control or miR-139-5p inhibitor, and 48 hr after transfection, the cells were treated with vehicle, 2 μM INZ-C, 5 nM ActD or 10 μM Nutlin-3 for 48 hr, followed by MTT assay to determine cell viability.
*p<0.05 as compared to the respective miRNA inhibitor control.
Figure 5 with 1 supplement
miR-139-5p is negatively correlated with PDE4D expression in human colorectal tumor samples, and represses the growth of SW480 xenograft tumors.
(A) miR-139-5p is negatively correlated with PDE4D expression in colon tumor samples. miR-139-5p (top panel) and PDE4D (middle panel) RNA expression was determined in 50 tumors (T) and paired …
Figure 5—figure supplement 1
Quantification of PDE4D and Ki67 IHC analysis.
The IHC staining intensity was categorized to low, medium and high as determined by ImageJ software. Five random fields were chosen from each slide to obtain average intensity of each category, and …
Author response image 1
Downregulation of PDE4D by p53 and miR-139-5p.
(A) H460 cells were treated with DMSO, 2 μM INZ-C or 0.5 μM Dox for 18 hr followed by qRT-PCR analysis of miR-139-5p. (B) and (C) H460 cells were transfected with pSIF-H1-Scramble control (SC) or …
Author response image 2
Knockdown of BIM rescue doxorubicin suppression of H460 cell proliferation.
H460 cells were transfected with two siRNAs against BIM, and 48 hr after transfection, cells were treated with 0.5 μM Dox for indicated duration and cell proliferation was determined by using the …
Author response image 3
miR-139-5p induces apoptosis.
H460 cells were transfected with pSIF-H1-Scramble control (Control) or pSIF-H1-miR-139-5p (139-5p), and 48 hr after transfection, the cells were subjected to flow cytometry analysis. *p<0.05. Error …
Additional files
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Supplementary file 1
List of primers used in this study.
- https://doi.org/10.7554/eLife.15978.018
