Wang Lin, Cao Hong, Pang Xueli, Li Kuangfa, Dang Weiqi, Tang Hao, Chen Tingmei
Key Laboratory of Laboratory Medical Diagnostics, Ministry of Eduction, Chongqing Medical University, Chongqing 400016, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2013 Dec;29(12):1272-6.
To investigate the effect and the relevant molecular mechanisms of leptin on the migration and invasion of human breast cancer MCF-7 cells.
The expression of OB-R in MCF-7 cells was measured by RT-PCR and Western blotting. The effects of leptin (100 ng/mL) on the the phosphorylation of a few key cell signaling proteins, p-ERK1/2, p-STAT3, p-AKT in MCF-7 cells were examined by Western blotting. Cell scratch assay and Transwell(TM); assay were utilized to measure the effects of leptin on the migration and invasion capability of MCF-7 cells, respectively. The effects of leptin on the mRNA and protein expression of matrix metalloproteinas 9 (MMP-9) and transforming growth factor β (TGF-β) were measured by RT-PCR and Western blotting.
Both OB-Rb and OB-Rt were expressed in MCF-7 cells. This indicated that leptin may have significant activities in MCF7 cells. Indeed, leptin increased the phosphorylation of p-ERK1/2, p-STAT3, and p-AKT in MCF-7 cells (P < 0.05). Further, leptin promoted migration and invasion of MCF-7 cells, which were attenuated by the JAK/STAT inhibitor AG490 (50 μmol/L), and the PI3K/AKT inhibitor LY294002 (10 μmol/L) (P < 0.05). Similarly, leptin also increased the mRNA and protein expression of MMP-9 and TGF-β, and these effects were blocked by AG490 and LY294002 as well (P < 0.05).
Leptin promoted the migration and invasion capabilities of MCF-7 cells. These activities may be achieved by the upregulation of MMP-9 and TGF-β through JAK/STAT and PI3K/AKT signaling pathways.
探讨瘦素对人乳腺癌MCF-7细胞迁移和侵袭的影响及其相关分子机制。
采用RT-PCR和蛋白质免疫印迹法检测MCF-7细胞中OB-R的表达。通过蛋白质免疫印迹法检测瘦素（100 ng/mL）对MCF-7细胞中几种关键细胞信号蛋白p-ERK1/2、p-STAT3、p-AKT磷酸化的影响。分别利用细胞划痕试验和Transwell试验检测瘦素对MCF-7细胞迁移和侵袭能力的影响。采用RT-PCR和蛋白质免疫印迹法检测瘦素对基质金属蛋白酶9（MMP-9）和转化生长因子β（TGF-β）mRNA和蛋白表达的影响。
MCF-7细胞中OB-Rb和OB-Rt均有表达。这表明瘦素可能在MCF-7细胞中具有显著活性。事实上，瘦素增加了MCF-7细胞中p-ERK1/2、p-STAT3和p-AKT的磷酸化水平（P < 0.05）。此外，瘦素促进了MCF-7细胞的迁移和侵袭，而JAK/STAT抑制剂AG490（50 μmol/L）和PI3K/AKT抑制剂LY294002（10 μmol/L）可减弱这些作用（P < 0.05）。同样，瘦素也增加了MMP-9和TGF-β的mRNA和蛋白表达，而AG490和LY294002也可阻断这些作用（P < 0.05）。
瘦素促进了MCF-7细胞的迁移和侵袭能力。这些作用可能是通过JAK/STAT和PI3K/AKT信号通路上调MMP-9和TGF-β来实现的。
