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目的观察瘦素对人乳腺癌细胞系 MCF-7增殖的影响,探讨其作用机制。方法免疫荧光检测人乳腺癌细胞系 MCF-7瘦素受体(OB-Rb)的表达。于 MCF-7细胞中分别加入不同浓度的瘦素(0、10、50、100、150μg/L),作用不同的时间(6、12、24 h),噻唑蓝(MTT)法检测各组细胞的增殖情况,同法分别检测 STAT3抑制剂 AG490和 ERK1/2抑制剂 PD98059对瘦素刺激 MCF-7细胞增殖的影响。采用免疫印迹法(Western blot)检测瘦素(100μg/L)作用于 MCF-7细胞不同时间(0、20、40、60 min)STAT3和 ERK1/2的磷酸化水平。结果免疫荧光检测证实 MCF-7细胞存在瘦素受体(OB-Rb)表达。瘦素能明显促进 MCF-7细胞的增殖,并呈时间和剂量依赖性,于100μg/L瘦素作用24 h 时增殖效应最大,100μg/L 组与150μg/L 组差异无统计学意义(P>0.05);不同浓度的 AG490与 PD98059均可明显抑制瘦素对 MCF-7细胞的增殖,随着 AG490与 PD98059浓度增高,抑制作用逐渐增强;MCF-7细胞经100ng/ml 瘦素处理后,随时间延长 STAT3和 ERK1/2磷酸化程度逐渐增高,且持续至少达60 min。结论瘦素可能通过激活 STAT3和 ERK 信号通路促进人乳腺癌细胞系 MCF-7的增殖。
Objective To observe the effect of leptin on the proliferation of human breast cancer cell line MCF-7 and to explore its mechanism. Methods The expression of leptin receptor (OB-Rb) in human breast cancer cell line MCF-7 was detected by immunofluorescence. Different concentrations of leptin (0, 10, 50, 100 and 150 μg / L) were added to MCF-7 cells at different time points (6, 12 and 24 h) The proliferation of MCF-7 cells was stimulated with leptin by AG490, a STAT3 inhibitor, and PD98059, an ERK1 / 2 inhibitor, respectively. The phosphorylation levels of STAT3 and ERK1 / 2 in MCF-7 cells treated with leptin (100 μg / L) at different time points (0, 20, 40 and 60 min) were determined by Western blotting. Results Immunofluorescence confirmed the expression of leptin receptor (OB-Rb) in MCF-7 cells. Leptin obviously promoted the proliferation of MCF-7 cells in a dose- and time-dependent manner. The effect of leptin at 100 μg / L was greatest at 24 h, and there was no significant difference between 100 μg / L and 150 μg / L groups (P > 0.05). Different concentrations of AG490 and PD98059 could significantly inhibit the proliferation of MCF-7 cells induced by leptin. With the increase of the concentration of AG490 and PD98059, the inhibition increased gradually. After treatment with 100ng / ml leptin, The phosphorylation of STAT3 and ERK1 / 2 gradually increased over time and lasted for at least 60 min. Conclusion Leptin may promote the proliferation of human breast cancer cell line MCF-7 by activating STAT3 and ERK signaling pathways.