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Chemical Structure| 154447-36-6 Chemical Structure| 154447-36-6

Structure of LY294002
CAS No.: 154447-36-6

Chemical Structure| 154447-36-6

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LY294002 is a PI3K inhibitor with IC50 values of 0.5 μM for PI3Kα, 0.57 μM for PI3Kδ, and 0.97 μM for PI3Kβ. LY294002 has anti-tumor effects and is used to study the PI3K/AKT signaling pathway and related cancer treatments.

Synonyms: SF 1101; NSC 697286

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Sharma, Pooja ; Tiufekchiev, Sarah ; Lising, Victoria ; Chung, Seung Woo ; Suk, Jung Soo ; Chung, Byung Min

Abstract: Cyclin D3 regulates the G1/S transition and is frequently overexpressed in several cancer types including breast cancer, where it promotes tumor progression. Here we show that a cytoskeletal protein keratin 19 (K19) phys. interacts with a serine/threonine kinase GSK3β and prevents GSK3β -dependent degradation of cyclin D3. The absence of K19 allowed active GSK3β to accumulate in the nucleus and degrade cyclin D3. Specifically, the head (H) domain of K19 was required to sustain inhibitory phosphorylation of GSK3βSer9, prevent nuclear accumulation of GSK3β,and maintain cyclin D3 levels and cell proliferation. K19 was found to interact with GSK3β and K19-GSK3β interaction was mapped out to require Ser10 and Ser35 residues on the H domain of K19. Unlike wildtype K19, S10A and S35A mutants failed to maintain total and nuclear cyclin D3 levels and induce cell proliferation. Finally, we show that the K19-GSK3β-cyclin D3 pathway affected sensitivity of cells toward inhibitors to cyclin-dependent kinase 4 and 6 (CDK4/6). Overall, these findings establish a role for K19 in the regulation of GSK3β-cyclin D3 pathway and demonstrate a potential strategy for overcoming resistance to CDK4/6 inhibitors.

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Sharma, Pooja ;

Abstract: Keratins are widely used as diagnostic markers to detect tumors in both primary and distal sites and to determine tumors’ tissue of origin in order to aid in treatment strategies. In this regard, K19 has been particularly useful because it is among the most sensitive diagnostic markers across a broad range of cancer types. K19 has also been shown to be one of the most reliable prognostic markers for multiple tumor types, including breast cancer where higher expression of K19 is correlated with worse patient survival. Despite the clinical data showing positive correlation between increased K19 expression and poor survival rates among patients of breast cancer types, the role of K19 in breast cancer remains unclear. To understand the role of K19 in breast cancer, we generated KRT19 knockout (KO) cell lines using MCF7 and MDA-MB-231 breast cancer cell lines. Using this system, we found that KRT19 KO cells exhibit reduced proliferation compared to parental control cells and uncovered a cell cycle promoting role of K19. We identified that K19 promotes proliferation of cancer cells by stabilizing a cell cycle regulator cyclin D3. However, how a cytoskeletal protein regulates levels of cyclin D3 remained unknown. It has been known that GSK3β play a crucial role in degrading cyclin D3 for proteasomal degradation. In our study in MCF7 cells, we identified GSK3β as a keratin-interacting protein and found that K19 suppressed GSK3β activity by inhibiting its nuclear accumulation. GSK3β-binding by K19 required serine 10 and 35 residue as K19 S10A or S35A mutation failed to protect cyclin D3 from protein degradation. Our results reveal a novel regulatory role of K19 on GSK3β localization and activity as a mechanism of how a cytoskeletal protein stabilizes a cell cycle regulator and promotes proper cell proliferation. In addition, we found that K19 expressing cells are more sensitive to cyclin dependent kinase (CDK) 4/6 inhibitors as compare to KRT19 KO cells. The sensitivity of KRT19 KO cells to CDK4/6 inhibitor was increased when cotreated with a GSK3β inhibitor, demonstrating that the K19-GSK3β axis plays a critical role in drug resistance. Given that K19 expression levels are frequently elevated in various cancers, K19 can be used to predict the efficacy of CDK4/6 inhibitors and patients may be cotreated with a GSK3β inhibitor to avoid drug resistance.

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Product Details of LY294002

CAS No. :154447-36-6
Formula : C19H17NO3
M.W : 307.34
SMILES Code : O=C1C=C(N2CCOCC2)OC3=C1C=CC=C3C4=CC=CC=C4
Synonyms :
SF 1101; NSC 697286
MDL No. :MFCD00270881
InChI Key :CZQHHVNHHHRRDU-UHFFFAOYSA-N
Pubchem ID :3973

Safety of LY294002

GHS Pictogram:
Signal Word:Warning
Hazard Statements:H302-H315-H319-H332-H335
Precautionary Statements:P261-P280-P305+P351+P338

Related Pathways of LY294002

DNA
PI3K-AKT
Hedgehog

Isoform Comparison

Biological Activity

Target
  • p110β

    p110β, IC50:0.97 μM

  • p110α

    p110α, IC50:0.5 μM

  • p110δ

    p110δ, IC50:0.57 μM

  • CK2

    CK2, IC50:98 nM

In Vitro:

Cell Line
Concentration Treated Time Description References
ADSCs 50 µM 24 h To investigate the role of PI3K signaling on rGO/PCL induced neurotrophic phenotype changes and myelin protein secretion. PMC9640298
H9C2 cells 25 μM 30 min To explore the mechanisms of NGR1's protective effect on H9C2 cells. PMC9287735
chondrocytes 20 μM 3 days The study aims to investigate the effects of blood on chondrocyte viability and apoptosis. PMC11076293
MLE-12 cells 50 μg/mL 24 h LY294002 was used as a positive control to inhibit up-regulated expression of fibrosis-related genes. PMC11515248
BMSCs 20 µM 0, 15, 30, 60, 120 min LY294002 significantly inhibited the higher expression of osteogenic genes induced by Ginsenoside Rb1. PMC8841501
CTLL-2 cells 50 μM 48 h The purpose was to analyze the effect of LY294002 on the CTLL-2 cells in the presence of IL-7. PMC10542607
Astrocytes 40 μM immediately post-OGD/R To inhibit PI3K signaling pathway and evaluate its effects on glycophagy. PMC11215420
hWJ-MSCs 40 µM 2 h Estimate the effects of the AKT inhibitor on the proliferation of FP2-cultured hWJ-MSCs. PMC11331723
RAW 264.7 10 µM 24 h To evaluate the effect of LY294002 on macrophage polarization. PMC11162076
SKMEL28 melanoma cells 20 µM 2 h The study involved the inhibition of AKT signaling to determine the effects on oxidative DNA damage repair; results showed reduced expression of OGG. PMC11002308
various TNBC cell lines 2 μM 10 days to investigate the effect of MAPK4 knockdown on cell growth and sensitivity to PI3K inhibitors PMC8752662
Human fetal RPE cells 2 μM 4 days To investigate the effect of glucose concentration on inflammatory responses, where inflammatory markers were evaluated. PMC9561713
NHA/HRasV12/TRIM24 cells 0.01 to 100 μM 24, 48, and 72 h Cell proliferation was evaluated and the half-maximal inhibitory concentration (IC50) values were determined from fitted concentration-response curves. PMC11304257
RF24 cells 50 µM 6 h The treatment resulted in reduced angiogenesis as inferred from subsequent experiments. PMC10133315
HAEC and HSMEC 50 nM 30 min To evaluate the effect of LY294002 on insulin uptake. PMC10435471
U-251 and Ca9-22 cells 20 μm 2 h To assess the impact of PI-3 kinase inhibition on cell behavior. PMC9982589
hESCs 10 µM 24 h To investigate the role of LY294002 in influencing differentiation to endoderm. PMC9876972
Astrocytes 10 µM 1 h AMPK inhibition reduces collective cell migration and invasion efficiency. PMC9372137
Vascular Smooth Muscle Cells (VSMC) 50 μM 30 min To inhibit the activation of Akt and study the effects on related signaling pathways. PMC8980891
Human embryonic stem cells 5 μM 8 days Induction of neuroectoderm differentiation. PMC11109972
HUVECs 5 μg/mL variable (6, 24, 48, and 72 h) to observe the effects of the PI3K inhibitor as a negative control PMC9972357
Primary chondrocytes 10 µM 24 h LY294002 administration significantly reduced SNP-induced apoptosis activities and alleviated aberrant force-induced chondrocyte degeneration. PMC9279410
RAW264.7 cells 25 µM 1 h Inhibit PI3K signaling pathway and reduce M1 macrophage polarization PMC11295337
BMDMs 25 µM 1 h Inhibit PI3K signaling pathway and reduce M1 macrophage polarization PMC11295337
neonatal rat cardiomyocytes (NRCMs) 10 μM 2 h Inhibited PI3K α/AKT signaling, reversed FGF7-mediated promotion of Nrf2 nuclear translocation, and increased ROS levels and apoptotic cells PMC9482143
human dental pulp stem cells 10 µM 7 days LY294002 attenuated the responsiveness of 10−6 M insulin to IIS/PI3K/AKT/mTOR pathway axis, suppressing the promoting effect of insulin on cell proliferation, osteogenic differentiation and bone formation. PMC11287875
PC-9R cells 25 μM Inhibited the PI3K/AKT pathway and restored sensitivity to gefitinib in PC-9R cells PMC8990424
BMDMs 0.5 μM 2 h Inhibited PI3K/Akt/Nrf2 signaling pathway and increased NLRP3/Caspase-1/GSDMD protein expression PMC10060206
A549 cells 10μM 1 h Inhibited PI3K/AKT signaling pathway, alleviated TGF-β1-induced epithelial-mesenchymal transition (EMT) PMC8959854
Huh7-SR cells 5 µM 1 h Inhibited PI3K/Akt pathway, reduced Nrf2 nuclear translocation, and increased ferroptosis level PMC8980859
HepG2-SR cells 5 µM 1 h Inhibited PI3K/Akt pathway, reduced Nrf2 nuclear translocation, and increased ferroptosis level PMC8980859
Mouse primary hepatocytes 10 μM 1.5 h To evaluate the relationship between PI3K/AKT pathway and ferroptosis, LY294002 pretreatment exacerbated ferroptosis PMC11495534

In Vivo:

Species
Animal Model
Administration Dosage Frequency Description References
Mice Experimental autoimmune hepatitis model Intraperitoneal injection 30 mg/kg Pretreatment 30 minutes before Inhibit PI3K signaling pathway and reduce FGF4's regulatory effect on M1 macrophages PMC11295337
C57BL/6 mice Myocardial infarction model (LAD ligation) Intraperitoneal injection 2.5 mg/kg From LAD operation to 7 days post-operation Inhibited PI3K α/AKT signaling, abolished FGF7-mediated regulation of Nrf2 and HXK2, and exacerbated myocardial oxidative stress and apoptosis PMC9482143
BALB/c nude mice PC-9R cell xenograft model Intraperitoneal injection 2.5 mg/kg Every three days for 12 days LY294002 significantly reduced tumor volume and enhanced the anti-tumor effect of gefitinib PMC8990424
C57BL/6 mice Acute lung injury model Injection 20 mg/kg 24 hours Inhibited PI3K/Akt/Nrf2 signaling pathway and increased NLRP3/Caspase-1/GSDMD protein expression PMC10060206
Mice Bleomycin-induced pulmonary fibrosis model Intraperitoneal injection 25 mg/kg Every other day for 21 days Inhibited PI3K/AKT signaling pathway, alleviated bleomycin-induced pulmonary fibrosis and EMT PMC8959854
NOD mice Diabetes model Intraperitoneal injection 75 mg/kg Daily for 3 days Inhibition of PI3K signaling pathway to observe its effect on CD4+ T cell migration PMC5101163
NOD-SCID mice Subcutaneous tumor model Intraperitoneal injection 25 mg/kg Twice a week for 4 weeks Inhibited PI3K/Akt pathway, slowed growth of FNDC5-overexpressing HCC cells, and increased ferroptosis level PMC8980859
C57 mice Liver warm ischemia/reperfusion model Intraperitoneal injection 10 mg/kg Single dose 36 h before liver ischemia followed by 6 h reperfusion To assess the role of PI3K/AKT pathway in hepatic I/R injury, LY294002 pretreatment abolished the protective effect of rmGas6 and exacerbated ferroptosis PMC11495534

Clinical Trial:

NCT Number Conditions Phases Recruitment Completion Date Locations
NCT02337309 Neuroblastoma Phase 1 Terminated(Low patient accrual... More >>) Less << - United States, California ... More >> Children's Hospital Los Angeles Los Angeles, California, United States, 90027-0700 UCSF Helen Diller Family Comprehensive Cancer Center San Francisco, California, United States, 94143 United States, Colorado Children Hospital of Colorado Aurora, Colorado, United States, 80045 United States, Georgia AFLAC Cancer Center and Blood Disorders Service of Children's Healthcare of Atlanta - Egleston Campus Atlanta, Georgia, United States, 30322 United States, Illinois University of Chicago Comer Children's Hospital Chicago, Illinois, United States, 60637 United States, Massachusetts Childrens Hospital Boston, Dana-Farber Cancer Institute. Boston, Massachusetts, United States, 02115 United States, Michigan C.S Mott Children's Hospital Ann Arbor, Michigan, United States, 48109 United States, North Carolina University of North Carolina Chapel Hill, North Carolina, United States, 27599 United States, Ohio Cincinnati Children's Hospital Medical Center Cincinnati, Ohio, United States, 45229-3039 United States, Texas Cook Children's Healthcare System Fort Worth, Texas, United States, 76104 United States, Washington Children's Hospital and Regional Medical Center - Seattle Seattle, Washington, United States, 98105 Less <<

Protocol

Bio Calculators
Preparing Stock Solutions 1mg 5mg 10mg

1 mM

5 mM

10 mM

3.25mL

0.65mL

0.33mL

16.27mL

3.25mL

1.63mL

32.54mL

6.51mL

3.25mL

 

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