慢性炎症和能量代谢因子对乳腺癌细胞系KISS-1表达的影响

目的下丘脑弓状核KISS-1神经元,是调控下丘脑-垂体-性腺轴功能的神经中枢。各种因子可能通过影响下丘脑细胞KISS-1表达而调控性腺轴功能。因此,本研究旨在评价慢性炎症因子脂多糖（lipopolysaccharide,LPS）,肿瘤坏死因子α（tumor necrosis factor alpha,TNF-α）,白介素-6（interleukin6,IL-6）、地塞米松和能量代谢相关因子胰岛素、胰高糖素样肽（glucagon-like peptide,GLP-1）对乳腺癌MCF-7细胞系KISS-1表达的影响。方法对MCF-7细胞系进行培养,测定KISS-1 mRNA和蛋白表达水平。在培养液中添加不同浓度LPS、TNF-α、IL-6、地塞米松、胰岛素和GLP-1刺激24 h、48 h和72 h,观察不同浓度的因子对KISS-1表达的影响。用实时聚合酶链反应（real-time polymerase chain reaction,RT-PCR）方法定量测定KISS-1 mRNA水平;用Western blotting方法半定量评价KISS-1蛋白表达变化。结果 1LPS（10μg/ml）和地塞米松（10-6 M和10-7 M）可抑制KISS-1 mRNA和蛋白表达;2TNF-α（100ng/ml）、IL-6（10 ng/ml,20 ng/ml）和胰岛素（0.1 mIU/ml,0.01mIU/ml）可增加KISS-1 mRNA和蛋白的表达;3GLP-1类似物（艾塞那肽,1 ng/ml和10 ng/ml）,在6 h一过性增加KISS-1mRNA和蛋白表达。结论慢性炎症和能量代谢相关因子,如LPS、TNF-α、IL-6、地塞米松、胰岛素和GLP-1影响MCF-7细胞系KISS-1表达,可能通过影响下丘脑细胞KISS-1表达,从而调控下丘脑-垂体-性腺轴的功能。

Effects of Inflammatory and Energy Homeostasis Factors on the Expression of KISS-1 in Breast Cancer Cell-line MCF-7

Objective To clarify the effects of inflammatory and energy homeostasis factors,such as lipopolysaccharide（LPS）,tumor necrosis factor alpha（TNF-α）,interleukin 6（IL-6）,dexamethasone,insulin and glucogan-like peptide（GLP-1）analogue,on the expression of KISS-1in breast cancer cell-line MCF-7.Methods The MCF-7 cells were cultured and the expression of KISS-1 were measured.The cells were stimulated by LPS,TNF-α,IL-6,dexamethasone,insulin,GLP-1 analogue with various concentrations at 24 h,48 h and 72 h.The expression of KISS-1 after been stimulated at mRNA and protein level were measured by real-time polymerase chain reaction（RT-PCR）and Western blotting respectively.Results LPS（10 ng/ml）and dexamethasone（10-6 M and 10-7 M）suppressed the expression of the KISS-1 at mRNA and protein level.TNF-α（100 ng/ml）,IL-6（10 ng/ml,20 ng/ml）and insulin（0.1 and 0.01 mIU/ml）increased the expression of KISS-1 at mRNA and protein level.GLP-1 analogue（1 ng/ml and10 ng/ml）transiently promote the expression of KISS-1.Conclusions Inflammatory and energy homeostasis factors,such as LPS,TNF-α,L-6,dexamethasone,insulin and GLP-1 analogue,can modulate KISS-1 expression in MCF-7 cells.These factors may modulate hypothalamus-pituitary-gonad axis function via the regulation of KISS-1 expression in hypothalamic neurons.