【摘要】：目的乳腺癌的發(fā)病率不斷上升,上皮-間質(zhì)轉(zhuǎn)化(epithelial to mesenchymal transition,EMT)在乳腺癌病變中廣泛存在。本研究探討TGF-β1在乳腺癌EMT中的作用及PTPN12在此過程中的作用和相關(guān)機制。方法蛋白質(zhì)印跡法檢測TGF-β1處理乳腺癌細(xì)胞后EMT相關(guān)基因以及PI3K、p-AKT和mTOR的表達;免疫組織化學(xué)檢測乳腺癌組織和癌旁組織中PTPN12表達;免疫共沉淀檢測PTPN12和p-AKT是否有直接相互作用;蛋白質(zhì)印跡法檢測轉(zhuǎn)染LV5-PTPN12后,E-Cadherin、Vimentin、Fibronectin、PI3K、p-AKT和mTOR蛋白的表達情況;裸鼠皮下成瘤檢測PTPN12對乳腺癌成瘤大小以及體積的影響。結(jié)果 NC組和TGF-β1組Vimentin蛋白表達量分別為(17.2±2.3)%和(79.7±8.3)%,t=12.5,P=0.008;Fibronectin蛋白表達量分別(17.4±2.3)%和(77.4±7.5)%,t=12.1,P=0.016。PTPN12在乳腺癌組織中表達明顯降低,PTPN12與乳腺癌病理分期分級以及腋窩淋巴結(jié)受累數(shù)目有關(guān),NC組和TGF-β1組PI3K蛋白表達量分別為(18.6±1.9)%和(76.3±5.9)%,t=12.1,P=0.029;p-AKT蛋白表達量分別為(27.6±3.2)%和(54.1±6.2)%,t=11.7,P=0.013;mTOR蛋白表達量分別為(35.2±3.2)%和(72.2±5.3)%,t=18.3,P=0.031;PTPN12和p-AKT有直接相互作用;PTPN12可以抑制TGF-β1誘導(dǎo)的EMT,LV5-PTPN12組和LV5-NC組Vimentin蛋白表達量分別為(17.2±2.3)%和(79.7±8.3)%,t=11.9,P0.018;Fibronectin蛋白表達量分別為(17.4±2.3)%和(77.4±7.5)%,t=10.2,P=0.019;E-Cadherin蛋白表達量分別為(82.1±8.4)%和(0.22±0.02)%,t=13.7,P0.001。PTPN12可以通過PI3K/AKT信號通路起作用,LV5-PTPN12組和LV5-NC組PI3K蛋白表達量分別為(13.2±1.2)%和(56.2±5.1)%,t=7.1,P=0.021;p-AKT蛋白表達量分別為(23.5±2.5)%和(77.1±6.3)%,t=9.2,P=0.008;mTOR蛋白表達量分別為(28.2±3.1)%和(71.7±6.1)%,t=12.8,P=0.032。體內(nèi)實驗表明,與對照組相比,LV5-PTPN12組荷瘤小鼠腫瘤體積和體質(zhì)量都明顯變小,LV5-PTPN12組和LV5-NC組腫瘤體積分別為(2.8±0.2)和(0.4±0.02)cm~3,t’=13.8,P0.001;體質(zhì)量分別為(3.1±0.3)和(0.4±0.02)g,t’=18.6,P0.001。結(jié)論 PTPN12在TGF-β1作用下通過PI3K/AKT信號通路誘導(dǎo)乳腺癌的EMT中起抑制作用。
[Abstract]:Objective the incidence of breast cancer is increasing and epithelial-interstitial transformation (epithelial to mesenchymal) is widely used in breast cancer. The purpose of this study was to investigate the role of TGF- 尾 1 in EMT of breast cancer and the role and mechanism of PTPN12 in this process. Methods the expression of EMT related genes, PI3KPP-AKT and mTOR in breast cancer cells treated with TGF- 尾 1 was detected by Western blot, PTPN12 expression in breast cancer tissues and adjacent tissues was detected by immunohistochemistry, and whether there was direct interaction between PTPN12 and p-AKT was detected by immunoprecipitation. The expression of PI3KTp-AKT and mTOR protein in LV5-PTPN12 was detected by Western blot, and the effect of PTPN12 on the size and volume of breast cancer was detected by subcutaneous tumorigenesis in nude mice. Results the expression of Vimentin protein in NC group and TGF- 尾 1 group were (17.2 鹵2.3)% and (79.7 鹵8.3)%, respectively. The expression of Vimentin protein was (17.4 鹵2.3)% and (77.4 鹵7.5)%, respectively. TGF-尾1緇凱I3K铔嬬櫧琛ㄨ揪閲忓垎鍒負(fù)(18.6鹵1.9)%鍜，

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