LY-294002 hydrochloride (NSC 697286) is a synthetic molecule inhibitor of PI3Kα/δ/β (IC50: 0.5/0.57/0.97 μM, in cell-free assays); more stable than Wortmannin in solution, and also is a blocker of autophagosome formation.

PI3Kδ:0.57μM, PI3Kβ:0.97 μM, PI3Kα:0.5μM

LY294002通过失活Akt/PKB，从而抑制细胞增殖并诱导凋亡。在这些结肠癌细胞系中，LY294002表现出显著的生长抑制和诱导凋亡效应，并降低磷酸化Akt（Ser473）[2]的表达。LY294002使肿瘤细胞的胞质体积减小，核固缩明显。此外，LY294002还特异性引起细胞生长G1期阻滞，几乎完全抑制黑色素瘤细胞的增殖，并部分抑制MG-63（成骨肉瘤细胞系）的增殖[3]。

LY294002能有效抑制肿瘤生长并诱导凋亡，特别是在LoVo肿瘤中表现出显著效果。因此，在小鼠腹膜炎癌症模型中展现出了卓越的效能[2]。LY294002显著地抑制卵巢癌的生长和腹水形成[3]。

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

Cell lines: Colon cancer cell lines DLD-1,LoVo,HCT15,and Colo205. Concentrations: 0–50 μM. Incubation Time: 0–48 hours. Method: 1.0×105 cells (100 μL volume/well) are inoculated into 96-well microtiter plates.LY294002 is added to triplicate wells and cultured at 37oC for 0–48 hours.After treatment,Premix WST-1 (10 μL) is added to each microculture well,and the plates are incubated for 60 minutes at 37oC,after which absorbance at 450 nm is measured with a microplate reader.[2]

Animal Models: Two groups of athymic nude mice (5–7 weeks) are inoculated i.p.with OVCAR-3 cells. Formulation: Dissolved in DMSO plus 0.25 ml of PBS. Dosages: 0–100 mg/kg. Administration: i.p.[3]