2A活性降低（2) 我們證明，netrin-1可以 增加PP2Ac的酪氨酸3硝基化修飾，從而活化PP2Ac3為了進(jìn)一步證明netrin-1通過(guò)增加 PP2A活性抑制MEK/ERK/c-Jun的活性并下調(diào)integrin P 4的表達(dá)，我們使用Apocynin處 理Miapaca II細(xì)胞，蛋白印跡檢測(cè)MEK2, ERK1/2和c-Jun的磷酸化以及integrin P 4的 表達(dá)水平。使用Apocynin抑制PP2A的活性同樣抑制netrin-1誘導(dǎo)的MEK/ERK信號(hào)活性 的降低，c-Jun磷酸化水平的降低和Integrin P 4表達(dá)的水平的降低。結(jié)果說(shuō)明PP2A在 netrin-1誘導(dǎo)Integrin P 4表達(dá)下降過(guò)程中起到非常關(guān)鍵的作用D
[0249] 13. netrin-1通過(guò)受體UNC5B降低MEK/ERK激酶活性并降低Integrin 0 4的表 達(dá)
[0250] 13. I netrin-1 通過(guò)受體 UNC5B 下調(diào) Integrin 0 4 的表達(dá)
[0251 ] 我們使用抗體阻斷的方法研究netrin-1的效應(yīng)受體。使用受體UNC5B， Neogeninl，Integrin a 3 P 1和Integrin a 6 P 4的抗體與細(xì)胞共同孵育12小時(shí)，之后 用netrin-1重組蛋白（20ng/ml)刺激阻斷后的細(xì)胞，通過(guò)蛋白印跡檢測(cè)MEK/ERK信號(hào)通 路和Integrin (64的表達(dá)水平的變化a結(jié)果表明，阻斷受體Integrin a 604，Integrin a 3 旦 1 和 Neogeninl 后，netrin-1 激活 FAK 活性，抑制 MEK2-ERK1/2 活性和下調(diào) Integrin 0 4的效應(yīng)不受影響，而阻斷受體UNC5B后，netrin-1產(chǎn)生的效應(yīng)消失。說(shuō)明netrin-1通 過(guò)其受體UNC5B下調(diào)Integrin 0 4的表達(dá)D
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【主權(quán)項(xiàng)】
1. netrin-1蛋白在制備用于腫瘤治療的藥物中的用途。2. 檢測(cè)netrin-1蛋白表達(dá)的試劑在制備腫瘤診斷試劑盒中的用途。3. 根據(jù)權(quán)利要求2所述的用途，其中所述試劑為netrin-1蛋白的抗體。4. 含有編碼netrin-1蛋白核酸的載體在制備用于腫瘤治療的藥物中的用途。5. 根據(jù)權(quán)利要求4所述的用途，其中所述含有編碼netrin-1蛋白核酸的載體為過(guò)表達(dá) netrin-1蛋白的載體。6. 根據(jù)前述權(quán)利要求中任一項(xiàng)所述的用途，其中所述腫瘤選自胰腺癌，優(yōu)選為胰導(dǎo)管 腺癌。7. 根據(jù)權(quán)利要求6所述的用途，其中所述腫瘤為I/II期胰導(dǎo)管腺癌，優(yōu)選為MIAPACA II細(xì)胞系。8. -種用于治療和/或抑制腫瘤的藥物組合物，其包含netrin-1蛋白或含有編碼 netrin-1蛋白核酸的載體以及一種或多種藥用賦形劑或藥用載體，所述腫瘤優(yōu)選為胰腺 癌，更優(yōu)選為胰導(dǎo)管腺癌。9. 根據(jù)權(quán)利要求8所述的用于治療和/或抑制腫瘤的藥物組合物，其包含過(guò)表達(dá) netrin-1蛋白的載體。10. 根據(jù)權(quán)利要求8所述的用于治療和/或抑制腫瘤的藥物組合物，其中所述 netrin-1的有效測(cè)試濃度為0-50ng/mL，優(yōu)選為10、20、30、40、50ng/mL，更優(yōu)選為10或 20ng/mL，最優(yōu)選為 20ng/mL。
【專利摘要】本發(fā)明公開了netrin-1蛋白在制備用于腫瘤治療的藥物中的用途。具體地，本發(fā)明公開了netrin-1蛋白在制備用于腫瘤治療的藥物中的用途，所述腫瘤選自胰腺癌，更優(yōu)選為胰導(dǎo)管腺癌。本發(fā)明的公開對(duì)于預(yù)防、診斷和治療胰導(dǎo)管腺癌以及開發(fā)治療胰導(dǎo)管腺癌的藥物具有積極意義。
【IPC分類】G01N21/31, A61K48/00, A61K38/17, G01N33/68, A61P35/00
【公開號(hào)】CN105031611
【申請(qǐng)?zhí)枴緾N201510341113
【發(fā)明人】呂湘, 劉德培, 安曦洲, 趙治國(guó)
【申請(qǐng)人】中國(guó)醫(yī)學(xué)科學(xué)院基礎(chǔ)醫(yī)學(xué)研究所
【公開日】2015年11月11日
【申請(qǐng)日】2015年6月18日