WAVE1对K562细胞侵袭能力的影响及其机制研究

目的 研究WASP家族富含脯氨酸同源蛋白1(WAVE1)对K562细胞侵袭的影响及其机制.方法 免疫荧光观察WAVE1与基质金属蛋白-2(MMP-2)在细胞中的分布.利用PeDNA3.1WAVE1重组真核表达质粒转染K562细胞,将WAVE1基因特异性小片段干扰RNA(WAVEI siRNA)转染K562细胞,Transwell法检测细胞侵袭能力;实时PCR和Western blot检测转染前后WAVE1及MMP-2的表达.结果 ①WAVEI与MMP-2主要表达于K562细胞的细胞膜上且两者有共定位.②与对照组K562细胞相比,转染pcDNA3.1-WAVE1 24 h及48 h后K562细胞MMP-2 mRNA表达分别增加295%和198%,蛋白表达水平分别增加80%和23%;转染特异性WAVE1 siRNA 24 h及48 h后K562细胞MMP-2 mRNA表达分别下降81%和28%,蛋白表达分别下调36%和53%.③与对照组K562细胞相比转染pcDNA3.1-WAVE1后细胞侵袭能力增强,而干扰WAVE1表达后K562细胞侵袭能力减弱.结论 WAVE1与MMP-2在K562细胞可能具有协同作用;WAVE1参与了K562细胞的侵袭转移过程,其机制可能与调控MMP-2的表达相关.

Role of WAVEI in K562 leukemia cells invasion and its mechanism

Objective To investigate role of WASP family verprolin homologous protein 1(WAVE1)in K562 leukemia cell invasion and its mechanism.Methods Immunofluoresence methed was used to detect the distribution of WAVE1 and MMP-2 in the cells.K562 cells were transfected with pcDNA3.1-WAVE1 reconstructed plasmid or with specific siRNA to WAVE1 gene.The invasion ability of K562 cells Was examined by Transwell assay.The expression level of WAVE1 and MMP-2 in K562 cells was assayed by real-time PCR by 295%and 198%while its protein increased by 80%and 23%respectively as compared with control K562cells.At the same time point ofter transfected with specific siRNA,the MMP-2 mRNA level decreased by vasion ability of K562 cells was enhanced after transfected with pcDNA3.1-WAVE1 and depressed after transfected with the specific siRNA.Conclusion The co-localization of WAVE1 and MMP-2 in K562 cells suggests they coordinate in functions:WAVE1 may involve in the migration and invasion of K562 cells through regulating the expression level of MMP-2.