LIMK1调控骨肉瘤细胞耐药的实验研究及机制分析  

The experimental study and mechanism of LIMK1 regulation of osteosarcoma cell resistance

作　　者：陈瑞 高奕瑶 杨建增[1] 荣小丽[1] 王岩[1] 

机构地区：[1]吉林大学中日联谊医院科学研究中心,吉林长春130033

出　　处：《中国实验诊断学》2018年第1期99-103,共5页Chinese Journal of Laboratory Diagnosis

基　　金：国家自然科学基金(30772488);国家自然科学基金(81172000);吉林省科技厅自然科学基金(20150101175JC)

摘　　要：目的探索LIMK1(LIM Kinase1)基因表达上调对人成骨肉瘤细胞耐药的影响,分析其可能的机制。方法应用阳离子脂质体介导的方法,将含有LIMK1激酶活性的重组质粒CFP-LIMK1-WT cDNA按不同浓度梯度瞬时转染人骨肉瘤细胞MG63,通过RT-PCR验证转染效率;采用CCK-8法检测上述细胞对不同浓度长春新碱的耐药情况,并通过Multi Experiment Matrix(MEM)和The Database for Annotation,Visualization and Integrated Discovery(DAVID)数据平台筛选LIMK1共表达基因及其富集信号通路,分析可能的耐药机制。结果随着转染浓度梯度增加,LIMK1及multidrug resistance protein 1(MDR1)基因表达呈明显上调趋势,人骨肉瘤细胞MG63的耐药程度增强。LIMK1及其共表达基因主要富集的信号通路有Rap1signaling pathway、TNF signaling pathway等。结论LIMK1基因表达上调促进人骨肉瘤细胞耐药,上述作用可能通过Rap1signaling pathway、TNF signaling pathway等信号通路来调节。Objective To investigate the effect of up regulation of LIMK1 （LIM Kinasel） gene expression on the drug resistance of human osteosarcoma cells and to analyze its possible mechanisms. Methods Application of cationic liposomal mediated method, the different concentrations of recombinant plasmid containing a LIMK1 kinase activity CFP LIMK1-WT cDNA by transient transfect into the human osteosarcoma cells MG63 ,the transfection efficiency was determined by RT- PCR. CCK- 8 method is used to detect the ability of these cells to resistant different concentrations of vincristine. The co- expressed genesof LIMK1 and those genes enrichment are analyzed by Multi Experiment Matrix （MEM） and The Database for Annotation, Visualization and Integrated Discovery（DAVID） respectively,to explore the possible mechanisms. Results With the increase of transfection concentration gradient, the expression of LIMK1 and multidrug resistance protein 1 （MDR1） gene increased significantly,and the drug resistance of human osteosarcoma cell line MG63 was also enhanced. The main enrichment signaling pathways of LIMK1 and its co- expressed genes include Rap1 signaling pathway,TNF signaling pathway and so on. Conclusion Up regulation of LIMK1 gene promotes drug resistance in human osteosarcoma cells, which may be regulated by the signaling pathways such as Rapl signaling path- way,TNF signaling pathway and so on.

关 键 词：LIMK1 骨肉瘤 多药耐药 

分 类 号：R738.1[医药卫生—肿瘤]