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目的:探讨MAPKs信号转导通路在瘦素(Leptin)促人乳腺癌细胞MCF-7增殖中的作用机制。方法:用MTT比色法观察不同浓度瘦素促MCF-7细胞的增殖效应,以及用JNK、ERK特异性抑制剂(SP600125,PD98059)阻断MAPKs信号通路对瘦素促MCF-7细胞增殖效应的影响;Westem blot检测瘦素干预不同时间对p-JNK、JNK、p-ERK、ERK蛋白表达水平的影响;RT-PCR检测在瘦素作用下以及MAPKs信号通路阻断情况下对MCF-7细胞表达瘦素受体(Ob-R)mRNA水平的影响。结果:50 ng/ml瘦素促人乳腺癌细胞MCF-7增殖效应最强,该浓度瘦素可使MAPKs信号通路中的靶蛋白发生磷酸化激活;50 ng/ml瘦素作用于MCF-7细胞30 s后p-ERK蛋白表达即显著增加,3 min后p-JNK蛋白表达亦显著增加;分别用SP600125和PD98059阻断JNK、ERK信号通路可显著抑制瘦素促MCF-7细胞的增殖效应.亦可抑制瘦素作用下MCF-7细胞Ob-R mRNA表达水平的增加。结论:瘦素促人乳腺癌细胞MCF-7增殖效应可能与激活MAPKs信号通路有关,亦调控MCF-7细胞中瘦素受体的表达,在肿瘤细胞周围形成瘦素自分泌环,促进乳腺癌的发生发展。
AIM: To investigate the role of MAPKs signal transduction pathway in the proliferation of human breast cancer cell line MCF-7 induced by leptin. Methods: MTT assay was used to observe the proliferation effect of different concentrations of leptin on MCF-7 cells and the effect of MAPKs signal pathway on the proliferation of MCF-7 cells treated with JNK and ERK inhibitor (SP600125, PD98059) The effect of leptin on the expression of p-JNK, JNK, p-ERK and ERK protein was detected by Westem blot. The effect of leptin on MAPKs signal pathway was detected by RT-PCR. The effect of cells expressing leptin receptor (Ob-R) mRNA levels. Results: The proliferation effect of 50 ng / ml leptin on human breast cancer cell line MCF-7 was strongest. The leptin at this concentration could phosphorylate the target protein in MAPKs signal pathway. After 30 s, the expression of p-ERK protein was significantly increased, and p-JNK protein expression was also increased 3 min later. Blocking JNK and ERK signaling pathways with SP600125 and PD98059 respectively significantly inhibited the proliferation of leptin-stimulated MCF-7 cells It also inhibited the increase of Ob-R mRNA expression in MCF-7 cells induced by leptin. CONCLUSION: The proliferation effect of leptin-induced human breast cancer cell line MCF-7 may be related to the activation of MAPKs signaling pathway. It also regulates the expression of leptin receptor in MCF-7 cells and forms leptin autocrine loop around the tumor cells to promote breast cancer The occurrence and development.