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MicroRNA-132/212 family enhances arteriogenesis after hindlimb ischaemia through modulation of the Ras-MAPK pathway.

Lei Z, van Mil A, Brandt MM, Grundmann S, Hoefer I, Smits M, El Azzouzi H, Fukao T, Cheng C, Doevendans PA, Sluijter JP - J. Cell. Mol. Med. (2015)

Bottom Line: Moreover, in in vitro pericyte-endothelial co-culture cell assays, overexpression of miR-132 and mir-212 in endothelial cells results in enhanced vascularization, as shown by an increase in tubular structures and junctions.Our results suggested that miR-132/212 may exert their effects by enhancing the Ras-Mitogen-activated protein kinases MAPK signalling pathway through direct inhibition of Rasa1, and Spred1.The miR-132/212 cluster promotes arteriogenesis by modulating Ras-MAPK signalling via direct targeting of its inhibitors Rasa1 and Spred1.

View Article: PubMed Central - PubMed

Affiliation: Division Heart and Lungs, Department of Cardiology, University Medical Center Utrecht, Utrecht, The Netherlands.

No MeSH data available.


Related in: MedlinePlus

miR-132/212 modulate Ras-MAPK signalling by suppressing Rasa1, Spred1 and Spry1 in HUVECs. (A) Experimental setting for quantitative measure of active ERK1/2 using Bio-plex phosopho-ERK1/2 assay. (B) A working model for miR-132/212 in modulation of Ras-MAPK pathway. (C) Quantification of Spred1, Spry1 or Rasa1 expression level after siRNA transfection against Spred1, Spry1 or Rasa1 in HUVECs, Values in the graph are shown as mean ± SEM, *** P < 0.001 n = 3. (D) Quantification of phosophorylated ERK1/2 level by Bio-plex pro phosopho-ERK1/2 set. Note the sustain ERK1/2 phosphorylation is prolonged after miR-132, 212 transfection or siRNA against Spred1, Spry1, Rasa1 or combinations of the three. (E) Modelling the decay of phosophorylated ERK1/2 level from D.
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fig05: miR-132/212 modulate Ras-MAPK signalling by suppressing Rasa1, Spred1 and Spry1 in HUVECs. (A) Experimental setting for quantitative measure of active ERK1/2 using Bio-plex phosopho-ERK1/2 assay. (B) A working model for miR-132/212 in modulation of Ras-MAPK pathway. (C) Quantification of Spred1, Spry1 or Rasa1 expression level after siRNA transfection against Spred1, Spry1 or Rasa1 in HUVECs, Values in the graph are shown as mean ± SEM, *** P < 0.001 n = 3. (D) Quantification of phosophorylated ERK1/2 level by Bio-plex pro phosopho-ERK1/2 set. Note the sustain ERK1/2 phosphorylation is prolonged after miR-132, 212 transfection or siRNA against Spred1, Spry1, Rasa1 or combinations of the three. (E) Modelling the decay of phosophorylated ERK1/2 level from D.

Mentions: Spred1, Spry1 and Rasa1 are known inhibitors of Ras-MAPK signalling and their inhibition can prolong Ras-MAPK signalling upon growth factor stimulation 30–33. We therefore tested whether miR-132/212 could prolong Ras-MAPK signalling by inhibiting Spred1, Spry1 and Rasa1 in HUVECs. Compared with siRNA controls and miR controls, overexpression of miR-132 and miR-212 or knockdown of Spred1, Spry1and Rasa1, indeed prolonged ERK1/2 phosphorylation (Fig.5A–D). By using a non-linear one-phase exponential decay model and interpolation of the time of phosphorylated ERK1/2T1/2 to reach 50%, we observed that T1/2 was prolonged both by overexpression of miR-132/212 and by knockdown of its targets (Fig.5E).


MicroRNA-132/212 family enhances arteriogenesis after hindlimb ischaemia through modulation of the Ras-MAPK pathway.

Lei Z, van Mil A, Brandt MM, Grundmann S, Hoefer I, Smits M, El Azzouzi H, Fukao T, Cheng C, Doevendans PA, Sluijter JP - J. Cell. Mol. Med. (2015)

miR-132/212 modulate Ras-MAPK signalling by suppressing Rasa1, Spred1 and Spry1 in HUVECs. (A) Experimental setting for quantitative measure of active ERK1/2 using Bio-plex phosopho-ERK1/2 assay. (B) A working model for miR-132/212 in modulation of Ras-MAPK pathway. (C) Quantification of Spred1, Spry1 or Rasa1 expression level after siRNA transfection against Spred1, Spry1 or Rasa1 in HUVECs, Values in the graph are shown as mean ± SEM, *** P < 0.001 n = 3. (D) Quantification of phosophorylated ERK1/2 level by Bio-plex pro phosopho-ERK1/2 set. Note the sustain ERK1/2 phosphorylation is prolonged after miR-132, 212 transfection or siRNA against Spred1, Spry1, Rasa1 or combinations of the three. (E) Modelling the decay of phosophorylated ERK1/2 level from D.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4549050&req=5

fig05: miR-132/212 modulate Ras-MAPK signalling by suppressing Rasa1, Spred1 and Spry1 in HUVECs. (A) Experimental setting for quantitative measure of active ERK1/2 using Bio-plex phosopho-ERK1/2 assay. (B) A working model for miR-132/212 in modulation of Ras-MAPK pathway. (C) Quantification of Spred1, Spry1 or Rasa1 expression level after siRNA transfection against Spred1, Spry1 or Rasa1 in HUVECs, Values in the graph are shown as mean ± SEM, *** P < 0.001 n = 3. (D) Quantification of phosophorylated ERK1/2 level by Bio-plex pro phosopho-ERK1/2 set. Note the sustain ERK1/2 phosphorylation is prolonged after miR-132, 212 transfection or siRNA against Spred1, Spry1, Rasa1 or combinations of the three. (E) Modelling the decay of phosophorylated ERK1/2 level from D.
Mentions: Spred1, Spry1 and Rasa1 are known inhibitors of Ras-MAPK signalling and their inhibition can prolong Ras-MAPK signalling upon growth factor stimulation 30–33. We therefore tested whether miR-132/212 could prolong Ras-MAPK signalling by inhibiting Spred1, Spry1 and Rasa1 in HUVECs. Compared with siRNA controls and miR controls, overexpression of miR-132 and miR-212 or knockdown of Spred1, Spry1and Rasa1, indeed prolonged ERK1/2 phosphorylation (Fig.5A–D). By using a non-linear one-phase exponential decay model and interpolation of the time of phosphorylated ERK1/2T1/2 to reach 50%, we observed that T1/2 was prolonged both by overexpression of miR-132/212 and by knockdown of its targets (Fig.5E).

Bottom Line: Moreover, in in vitro pericyte-endothelial co-culture cell assays, overexpression of miR-132 and mir-212 in endothelial cells results in enhanced vascularization, as shown by an increase in tubular structures and junctions.Our results suggested that miR-132/212 may exert their effects by enhancing the Ras-Mitogen-activated protein kinases MAPK signalling pathway through direct inhibition of Rasa1, and Spred1.The miR-132/212 cluster promotes arteriogenesis by modulating Ras-MAPK signalling via direct targeting of its inhibitors Rasa1 and Spred1.

View Article: PubMed Central - PubMed

Affiliation: Division Heart and Lungs, Department of Cardiology, University Medical Center Utrecht, Utrecht, The Netherlands.

No MeSH data available.


Related in: MedlinePlus