LY294002 is an inhibitor of PI3K (IC50s: 0.5/0.57/0.97 μM for PI3Kα/δ/β in cell-free assays). It also blocks autophagosome formation.

LY294002 (5 μM) was shown to block insulin-induced phosphorylation of PKB Ser473 in CHO-IR cells [1]. When the cells were incubated in medium containing 20 μM LY294002 for 48 h, cell proliferation was remarkably decreased (>80%) in LoVo and Colo205 cells. LY294002 (10–50 μM), when added for 24 h, significantly decreased the population of DLD-1 cells in a concentration-dependent manner [2]. When the cells were cultured in medium containing different concentrations of LY294002 (0, 10, 25, 50, and 75 μM)for 24 h and 48 h, cell proliferation was remarkably decreased in a dose-dependent fashion [3].

The i.v. administration of LY294002 to mice in group A significantly suppressed the growth of s.c. tumors derived from both DLD-1 and LoVo cells, with a mean volume of 72% (range, 48–82%) and 55% (range, 40–61%), respectively [2]. Treatment with LY294002 (50 mg/kg, 75 mg/kg) significantly reduced mean NPC tumor burden as compared with the control group. Treatment with 10 mg/kg or 25 mg/kg LY294002 was less effective in decreasing tumor burden [3].

PI3K inhibition by LY294002 was determined in a radiometric assay using purified, recombinant enzymes (class IA and class IB) with 1 μM ATP. The kinase reaction was carried out for 1 h at room temperature (24 °C) and was terminated by addition of PBS. IC50 values were subsequently determined using a sigmoidal dose-response curve fit (variable slope). CK2 and GSK3β (glycogen synthase kinase 3β) inhibition were established by kinase selectivity screening. Inhibitor (10 μM; LY294002) was tested against the Upstate panel of kinases in 10 μM ATP [4].

The cells were seeded into 96-well plates at 5000 cells/well. Twenty-four hours after cells were seeded, the medium was removed and replaced in the presence of LY294002 (0 μmol/L, 10 μmol/L, 25 μmol/L, 50 μmol/L, and 75 μmol/L) dissolved in DMSO or DMSO only for an additional 24 h and 48 h. To avoid any nonspecific toxic effects of DMSO on cell growth, DMSO concentrations were maintained at 0.5% in all experiments. MTT dye (5 mg/mL) was added to each well. The reaction was stopped by the addition of DMSO, and optical density was measured at 490 nm on a multiwell plate reader. Background absorbance of the medium in the absence of cells was subtracted. All samples were assayed in triplicate, and the mean for each experiment was calculated. Results were expressed as a percentage of control, which was considered to be 100% [3].

Athymic nude mice were used when they were 6-8 weeks. Mice were randomly divided into free separated into five groups (n = 4 mice). Mice were housed in the same environment with controlled temperature, humidity, and a 12 h light/dark cycle. Mice were inoculated subcutaneously with CNE-2Z cells (1 × 10^6 cells/mouse in 200 μl of RPMI-1640) into the flank. The tumor take rate was 100%. After 1 week, an intraperitoneal injection was performed to the xenograft mice with different dosage of LY294002 (10 mg/kg, 25 mg/kg, 50 mg/kg, and 75 mg/kg twice weekly (n = 4 mice), each group for 4 weeks. Treated mice have monitored any signs. Body weight and tumors size were measured twice a week. Tumor size was measured using calipers and tumor volume was calculated (volume = long axis × short axis^2). At the end of the treatment, all mice were euthanized. One part of tumor tissue was fixed in formalin and embedded in paraffin, and another part was stored at -70°C [3].

154447-36-6

C19H17NO3

307.349

NSC 697286;LY 294002;SF 1101;LY294002

[1]Li X, Hu X, Tian G G, et al. C89 Induces Autophagy of Female Germline Stem Cells via Inhibition of the PI3K-Akt Pathway In Vitro[J]. Cells. 2019, 8(6): 606. [2]Zhao Y, Guo W, Gu X, et al. Repression of deoxynivalenol-triggered cytotoxicity and apoptosis by mannan/β-glucans from yeast cell wall: Involvement of autophagy and PI3K-AKT-mTOR signaling pathway[J]. International Journal of Biological Macromolecules. 2020, 164: 1413-1421. [3]Chaussade C, et al. Evidence for functional redundancy of class IA PI3K isoforms in insulin signalling. Biochem J. 2007 Jun 15;404(3):449-58. [4]Semba S, et al. The in vitro and in vivo effects of 2-(4-morpholinyl)-8-phenyl-chromone (LY294002), a specific inhibitor of phosphatidylinositol 3'-kinase, in human colon cancer cells. Clin Cancer Res. 2002 Jun;8(6):1957-63. [5]Gharbi SI, et al. Exploring the specificity of the PI3K family inhibitor LY294002. Biochem J. 2007 May 15;404(1):15-21. [6]Wen Y, Peng D, Li C, et al. A new polysaccharide isolated from Morchella importuna fruiting bodies and its immunoregulatory mechanism[J]. International Journal of Biological Macromolecules. 2019, 137: 8-19. [7]Gu L, Ye P, Li H, et al. Lunasin attenuates oxidant-induced endothelial injury and inhibits atherosclerotic plaque progression in ApoE?/? mice by up-regulating heme oxygenase-1 via PI3K/Akt/Nrf2/ARE pathway[J]. The FASEB Journal. 2019 Apr;33(4):4836-4850. [8]Gu L, Wang Y, Xu Y, et al. Lunasin functionally enhances LDL uptake via inhibiting PCSK9 and enhancing LDLR expression in vitro and in vivo[J]. Oncotarget. 2017 Aug 24;8(46):80826-80840. [9]Chen Y, Sun Y, Zhao W, et al. Elevated SRC3 expression predicts pemetrexed resistance in lung adenocarcinoma[J]. Biomedicine & Pharmacotherapy. 2020, 125: 109958. [10]Jiang H, et al. Phosphatidylinositol 3-kinase inhibitor(LY294002) induces apoptosis of human nasopharyngeal carcinoma in vitro and in vivo. J Exp Clin Cancer Res. 2010 Apr 22;29:34.

[1]Liu H, Liu Y, Wang H, et al. Geometric Constraints Regulate Energy Metabolism and Cellular Contractility in Vascular Smooth Muscle Cells by Coordinating Mitochondrial DNA Methylation. Advanced Science. 2022: 2203995. [2]Zhang W, Yang S, Chen D, et al. SOX2-OT induced by PAI-1 promotes triple-negative breast cancer cells metastasis by sponging miR-942-5p and activating PI3K/Akt signaling. Cellular and Molecular Life Sciences. 2022, 79(1): 1-16. [3]Liu Y, Wang J, Chen J, et al. Upregulation of miR-520c-3p via Hepatitis B Virus Drives Hepatocellular Migration and Invasion through the PTEN/AKT/NF-κB Signaling Pathway. Molecular Therapy-Nucleic Acids. 2022 [4]Zhang H, Xia P, Liu J, et al. ATIC inhibits autophagy in hepatocellular cancer through the AKT/FOXO3 pathway and serves as a prognostic signature for modeling patient survival. International Journal of Biological Sciences. 2021, 17(15): 4442-4458. [5]Wang J, Li Y, Wan C M, et al. PTEN inhibition leads to the development of resistance to novel isoquinoline derivative TNBG-5602 in human liver cancer cells. American Journal of Cancer Research. 2021, 11(9): 4515. [6]Li X, Hu X, Tian G G, et al. C89 Induces Autophagy of Female Germline Stem Cells via Inhibition of the PI3K-Akt Pathway In Vitro. Cells. 2019, 8(6): 606 [7]Bai G, Wang H, Cui N. mTOR pathway mediates endoplasmic reticulum stress-induced CD4+ T cell apoptosis in septic mice. Apoptosis. 2022: 1-11 [8]Gu L, Wang Y, Xu Y, et al. Lunasin functionally enhances LDL uptake via inhibiting PCSK9 and enhancing LDLR expression in vitro and in vivo. Oncotarget. 2017 Aug 24;8(46):80826-80840 [9]Zhao Y, Guo W, Gu X, et al. Repression of deoxynivalenol-triggered cytotoxicity and apoptosis by mannan/β-glucans from yeast cell wall: Involvement of autophagy and PI3K-AKT-mTOR signaling pathway. International Journal of Biological Macromolecules. 2020, 164: 1413-1421 [10]Wen Y, Peng D, Li C, et al. A new polysaccharide isolated from Morchella importuna fruiting bodies and its immunoregulatory mechanism. International Journal of Biological Macromolecules. 2019, 137: 8-19.

H2O:<1 mg/mL
DMSO:34 mg/mL (110.6 mM)
Ethanol:10 mg/mL(32.5 mM)
Powder: -20°C for 3 years
In solvent: -80°C for 2 years