氯化锂抑制乳腺癌MDA-MB-231细胞侵袭的机制研究  

Mechanism of Lithium Chloride in Inhibiting Invasion of Breast Cancer MDA-MB-231 Cells

作　　者：王立洪[1] 李庆华[1] 王建[1] 高伟[1] 蔺亚妮[1] 李华文[1] 金薇娜[1] 常国强[1] 庞天翔[1] 

机构地区：[1]北京协和医学院血液学研究所血液病医院,中国医学科学院,实验血液学国家重点实验室,天津市300020

出　　处：《中国肿瘤临床》2011年第9期492-496,共5页Chinese Journal of Clinical Oncology

基　　金：天津市自然科学基金(编号:08JCZDJC19100;09JCZDJC17300)资助~~

摘　　要：目的:探讨LiCl对乳腺癌细胞系MDA-MB-231侵袭能力的影响,及其对侵袭关键因子NGAL表达的调控。方法:LiCI处理后,MTT测定细胞活力,光镜观察细胞伪足形成;Transwell观察细胞侵袭能力的变化;实时定量PCR及Western blotting观察NGAL在转录和翻译水平的表达;使用小分子药物Bay117082(NF-KB通路抑制剂)和FH535(Wnt/β-catenin通路抑制剂)处理细胞,检测关键通路对NGAL表达的调控。结果:在浓度低于10mmol/L时,LiCl对细胞活性的影响无显著性差异。5mmol/L或10mmol/LLiCl处理细胞24 h后,细胞形态发生明显改变,其伪足形成受到影响,但在NHEl抑制剂Cariporide作用下,细胞伪足破坏的情况有所减轻;Transwell结果显示5、10mmol/L LiCl分别处理细胞24 h后,细胞的侵袭能力受到明显抑制。实时定量PCR和Western blotting结果显示,LiCl处理后NGALmRNA和蛋白表达均明显下降,表明LiCl对NGAL的调控可能始于转录水平。小分子药物处理分别使Wnt/β-catenin和NF-kB信号通路抑制后,NGAL的表达均受到明显抑制,表明这两条通路都参与了NGAL表达的调控。结论:LiCl通过抑制NF-kB调控NGAL的表达,抑制伪足形成以及细胞侵袭。Objective： To investigate the effect of lithium chloride （LiCl） treatment on the invasion of breast cancer cell line MDA-MB-231, and its corresponding control on the expression of neutrophil gelatinase associated lipocalin （ NGAL ）, a key molecule responsible for the regulation of breast tumor cell metastasis. Methods： After LiCl treatment at various concentrations, the cell viability of MDA-MB-231 was determined by MTT. Meanwhile, the cell pseudopod formation was observed through inverted optical microscope, and changes in the capacity of cell invasion was analyzed by Transwell method, with pretreatment by a layer of Matrigel. The expression of NGAL mRNA and protein was detected by quantitative real time PCR and Western blot, respectively. Finally, small pharmacological molecules FH535 （ inhibitor for Wnt/β-catenin signaling pathway ） and Bay117082 （ inhibitor for NF-κB signaling pathway ） were used to analyze the important signaling pathways involved in the regulation of NGAL expression. Results： The effect of LiCl on the cell viability of MDA-MB-231 cell line was not statistically significant when the LiCl concentration was lower than 10 mmol/L. Representative pictures from the inverted microscope showed that cell pseudopod formation was inhibited after treatment with 5 mmol/ L or 10 mmol/L LiCl for 24h. However, this effect was somewhat reversed by another selective inhibitor Cariporide for Na＋/H＋ exchanger 1 （ NHE1 ）. The analysis of Transwell indicated that the capacity of cell invasion was apparently inhibited by LiCl, and the NGAL mRNA and protein expression was reduced by the same treatment. LiC1 could regulate the NGAL expression after the transcription. Treatment with inhibitors for Wnt/β-catenin and NF-KB signing pathways for 24h respectively dramatically downregulated the expression of NGAL protein. The results indicated that both of Wnt/β-catenin and NF-κB signaling pathways were involved in regulating the expression of NGAL. Conclusion： LiC1 can inhibit the pseu

关 键 词：氯化锂 乳腺癌 MDA-MB-231细胞 中性粒细胞明胶酶相关载脂蛋白 侵袭 

分 类 号：R737.9[医药卫生—肿瘤]