| 名稱 | LY294002 |
| 描述 | LY294002 (SF 1101) is a broad-spectrum inhibitor of PI3K, inhibiting PI3Kα, PI3Kδ, and PI3Kβ (IC50=0.5/0.57/0.97 μM). LY294002 is also an inhibitor of DNA-PK (IC50=1.4 μM) and an inhibitor of CK2 (IC50=98 nM). LY294002 activates apoptosis and autophagy. |
| 細(xì)胞實(shí)驗(yàn) | 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]. |
| 激酶實(shí)驗(yàn) | 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]. |
| 動(dòng)物實(shí)驗(yàn) | 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]. |
| 體外活性 | 方法：人胰腺癌細(xì)胞 AsPC-1、BxPC-3 和 PANC-1 用 LY294002 (5-45 μM) 處理 24 h，使用 MTT 方法檢測(cè)細(xì)胞生長(zhǎng)抑制情況。
結(jié)果：LY294002 劑量依賴誘導(dǎo) AsPC-1、BxPC-3 和 PANC-1 細(xì)胞生長(zhǎng)，IC50 分別為 40 μM、5 μM 和 35 μM。[1]
方法：表達(dá)人胰島素受體的中國(guó)倉(cāng)鼠卵巢細(xì)胞 CHO-IR 用 LY294002 (5 μM) 處理 5 min，并用 Insulin (1 nM, 10 min) 刺激，使用 Western Blot 方法檢測(cè)靶點(diǎn)蛋白表達(dá)水平。
結(jié)果：LY294002 可阻斷 CHO-I R細(xì)胞中胰島素誘導(dǎo)的 PKB Ser473 磷酸化。[2]
方法：人鼻咽癌細(xì)胞 CNE-2Z 用 LY294002 (10-75 μmol/L) 處理 24-48 h，使用 Flow Cytometry 方法檢測(cè)細(xì)胞凋亡情況。
結(jié)果：LY294002 劑量依賴性誘導(dǎo) CNE-2Z 細(xì)胞凋亡。[3] |
| 體內(nèi)活性 | 方法：為檢測(cè)體內(nèi)抗腫瘤活性，將 LY294002 (25 mg/kg，每周兩次) 和 cisplatin (5 mg/kg，每周一次) 腹腔注射給攜帶人胰腺癌腫瘤 AsPC-1 的 BALB/C nu/nu 小鼠，持續(xù)三周。
結(jié)果：對(duì)照組小鼠的腫瘤體積增加，而 cisplatin 或 LY294002 治療組的腫瘤體積分別減少為 77% 和 70%。聯(lián)合治療組更有效，腫瘤體積的生長(zhǎng)下降到對(duì)照組體積的 44%。[4]
方法：為研究 PI3K 的藥物阻斷是否能改善 LPS 誘導(dǎo)的小鼠急性肝損傷的發(fā)展，將 LY294002 (40 μM; 10 μL) 單次腹腔注射給 LPS 誘導(dǎo)的急性肝損傷 BALB/c 小鼠模型。
結(jié)果：LPS 誘導(dǎo)的肝炎中，LY294002 治療明顯抑制了各種疾病相關(guān)促炎細(xì)胞因子的肝內(nèi)合成，包括腫瘤壞死因子-α、IL-6、IL-1β 和 INF-γ。在 LPS 損傷的小鼠肝臟樣本中，觀察到 LY294002 顯著抑制 IκB 磷酸化。因此，LY294002 可能通過(guò)抑制活化的 B 細(xì)胞依賴性信號(hào)通路的 IκB 核因子 κ 輕鏈增強(qiáng)子來(lái)保護(hù)肝臟免受 LPS 誘導(dǎo)的損傷。[5] |
| 存儲(chǔ)條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | Ethanol : 10 mg/mL (32.5 mM)
10% DMSO+40% PEG300+5% Tween 80+45% Saline : 3.4 mg/mL (11.06 mM), Please add co-solvents sequentially, clarifying the solution as much as possible before adding the next one. Dissolve by heating and/or sonication if necessary. Working solution is recommended to be prepared and used immediately.
H2O : < 1 mg/mL (insoluble or slightly soluble)
DMSO : 61.46 mg/mL (200 mM), Sonication is recommended.
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| 關(guān)鍵字 | NSC-697286 | Lysophosphatidic acid | Leptin | NPC | NSC697286 | PI3K | DNA-PK | broad-spectrum | Autophagy | Phosphoinositide 3-kinase | DNA-dependent protein kinase | CNE-2Z | SF1101 | reversibly | Inhibitor | inhibit | YAP | translocation | Casein Kinase | LY-294002 | carcinoma | LY294002 | nasopharyngeal | spermatozoa | Human | Apoptosis | nuclear | tumor | SF-1101 |
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