赖氨酰氧化酶样基因-3在体外培养的人主动脉平滑肌细胞的表达

Expression of Human Lysyl Oxidase-Like Gene 3 in Human Aortic Smooth Muscle Cells

目的研究赖氨酰氧化酶样基因-3(Lysyl Oxidase-Like Gene 3,LOXL3)在体外培养的人主动脉平滑肌细胞(Hu-man Aortic Smooth Muscle Cell,HAoSMC)的表达模式。方法提取不同生长阶段的人主动脉平滑肌细胞的不同组分,用Northern Blot、Western Blot从mRNA和蛋白水平分析LOXL3表达,并与赖氨酰氧化酶样基因-2(Lysyl Oxidase-Like Gene 2,LOXL2)比较。结果LOXL3 mRNA和蛋白含量在细胞汇合前高于细胞汇合期和汇合后,且LOXL3蛋白在细胞外基质和细胞的不可溶组分中含量较高,而LOXL2则在细胞培养液中表达较高。结论LOXL3的表达量在细胞生长的不同阶段有所不同,人主动脉平滑肌细胞的LOXL3和LOXL2分布不同,可能与其功能有关。

Objective To investigate the expression pattern of Human Lysyl Oxidase-Like Gene 3（LOXL3） in cultured human aortic smooth muscle cells at different stages of confluency. Methods The expression pattern of LOXL3 was detected by Northem and Western blot analyses, and compared with the expression pattern of Lysyl Oxi- dase-Like Gene 2（LOXL2）. Results At different stages of cell culture, Northern and Western blot analyses showed a higher steady-state LOXL3 mRNA and protein level at pre-confluency than confluency and 3 days postconfluency. Western blot analysis showed that LOXL3 was predominantly localized in extracellular matrix （ECM） and insoluble cell layer fractions, compared with LOXL2, which was primarily detected in cultured cell media （CCM）. Conclusion These data demonstrated that the LOXL3 had different temporal distribution in human aortic smooth muscle cells, and had different localization compared with LOXL2. This might implicate the unique functions of LOXL3 in LOX family.