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Updated: May 9, 2026
Analysis of Retinoic Acid-induced Neural Differentiation of Mouse Embryonic Stem Cells in Two and Three-dimensional Embryoid Bodies
Published on: April 22, 2017
Src Family Kinase Inhibitor PP2 Has Different Effects on All-Trans-Retinoic Acid or Arsenic Trioxide-Induced
Suk-Gu Yoon1, Hee-Jeong Cheong, Sook-Ja Kim
1Division of Hematology-Oncology, Department of Internal Medicine, Institute for Clinical Molecular Biology Research, Soonchunhang University Hospital, Soonchunhang University College of Medicine, Seoul, Korea.
Purpose:
Leukemic promyelocytes have the unique ability to undergo differentiation after exposure to retinoic acid and both differentiation and apoptosis after exposure to arsenic trioxide (ATO). Recent studies have shown that inhibition of Src family kinases (SFKs) resulted in enhancement of retinoic acid-induced myeloid differentiation.
Materials And Methods:
In this study, we investigated the question of whether the SFK inhibitor PP2 enhanced the differentiation of NB4 cells when combined with ATO as well as when combined with all-trans-retinoic acid (ATRA). In addition, we attempted to determine the difference in retinoic acid-induced gene expression between cells treated with PP2 in combination with ATRA and in combination with ATO.
Results:
SFK inhibitor PP2 induced significant enhancement of ATRA- or ATO-induced differentiation of NB4 cells. A significantly stronger synergistic effect was observed when PP2 was combined with ATRA than when combined with ATO. Flow cytometric analysis demonstrated a significant increase in CD11b-positive granulocytes up to 60.73% and 31.58%, respectively. These results were confirmed by nitroblue tetrazolium staining. These effects were not related to apoptosis. Results of Annexin-V-fluorescein staining revealed that PP2 combined with ATRA or PP2 combined with ATO did not induce apoptosis in NB4 cells. Retinoic acid-induced gene expression was different in both groups. Intercellular adhesion molecule-1 expression showed a significant increase in cells treated with PP2 in combination with ATRA, whereas cathepsin D expression showed a significant increase in cells treated with PP2 in combination with ATO.
Conclusion:
Our data showed that SFK inhibitor PP2 enhanced acute promyelocytic leukemia cell differentiation when combined with either ATRA or ATO with difference in activation of retinoic acid-induced genes.
Insights
Src family kinase (SFK) inhibitor PP2 enhances acute promyelocytic leukemia cell differentiation with all-trans-retinoic acid (ATRA) or arsenic trioxide (ATO). PP2 combined with ATRA showed a stronger synergistic effect than with ATO, impacting gene expression differently.
Area of Science:
- Hematology
- Molecular Biology
- Cancer Research
Background:
- Leukemic promyelocytes differentiate with retinoic acid and differentiate/apoptose with arsenic trioxide (ATO).
- Inhibition of Src family kinases (SFKs) enhances retinoic acid-induced myeloid differentiation.
Purpose of the Study:
- To investigate if the SFK inhibitor PP2 enhances NB4 cell differentiation with ATO or all-trans-retinoic acid (ATRA).
- To determine differences in retinoic acid-induced gene expression when PP2 is combined with ATRA versus ATO.
Main Methods:
- NB4 cells were treated with PP2 in combination with ATRA or ATO.
- Differentiation was assessed by flow cytometry for CD11b expression and nitroblue tetrazolium staining.
- Apoptosis was evaluated using Annexin-V-fluorescein staining.
- Retinoic acid-induced gene expression was analyzed.
Main Results:
- PP2 significantly enhanced ATRA- or ATO-induced NB4 cell differentiation, with a stronger synergistic effect observed with ATRA.
- CD11b-positive granulocytes increased significantly (up to 60.73% with ATRA, 31.58% with ATO).
- No apoptosis was induced by PP2 in combination with ATRA or ATO.
- Intercellular adhesion molecule-1 (ICAM-1) expression increased with PP2+ATRA, while cathepsin D increased with PP2+ATO.
Conclusions:
- SFK inhibitor PP2 enhances acute promyelocytic leukemia cell differentiation induced by ATRA or ATO.
- The combination of PP2 with ATRA demonstrated a more pronounced synergistic effect on differentiation compared to ATO.
- Distinct retinoic acid-induced gene expression patterns were observed between the PP2+ATRA and PP2+ATO treatment groups.
