LPXN过表达对THP-1细胞增殖、粘附和侵袭的影响及其机制研究

摘要

目的:探讨LPXN过表达对白血病细胞株THP-1增殖、粘附和侵袭的影响及其可能机制.方法:构建稳定过表达LPXN的THP-1细胞.CCK-8检测细胞增殖能力;粘附实验检测细胞对Fn的粘附能力;Transwell实验检测细胞侵袭能力;Westem blot检测ERK,Integrinα4,α5和β1表达;明胶酶谱法检测细胞分泌MMP2和MMP9活性的变化.结果:成功构建稳定过表达LPXN的THP-1细胞.过表达LPXN的THP-1细胞增殖、对Fn的粘附、ERK,Integrinα4,α5和β1表达均高于对照组.过表达LPXN的THP-1细胞跨transwell的侵袭能力和MMP2活性均强于对照组.结论:LPXN基因可能通过上调ERK的表达促进THP-1细胞增殖;通过上调Integrin α4、α5、β1的表达促进THP-1细胞的粘附;通过激活MMP2促进THP-1细胞侵袭.%Objective:To explore the effect of LPXN overexpression on the proliferation,adhesion and invasion of THP-1 cells and its possible mechanism.Methods:A THP-1 cell line with stable overexpression of LPXN was constucted by using a lentivirus method,CCK-8 was used to detect the proliferation of cells,adhesion test was used to evaluate adhesion ablity of cells to Fn.Transwell assay was used to detect the change of invasion capability.Western blot was used to detect expression of LPXN,ERK,pERK and integrin α4,α5,β1,the Gelatin zymography was applied to detect activity of MMP2/MMP9 secreted by the THP-1 cells.Results:Successful establishment of THP-1 cells with LPXN overexpression (THP-1 LPXN) was confirmed with Western blot.THP-1 LPXN cells were shown to proliferate faster than the control THP-1 vector cells.Adhesion to Fn and expression of ERK,integrin α4,α5 and β1 in the THP-1 LPXN cells were higher than that in the control cells.Invasion across matrigel and enhanced activity of MMP2 could be detected both in the THP-1 LPXN cells as compared with the control cells.Conclusion:Ectopically ovexpression of LPXN may promote proliferation of THP-1 cells through up-regulation of ERK;promote adhesion of THP-1 cells through up-regulating the integrin α4/β1 as well as integrin α5/β1 complex;promote invasion of THP-1 cells through activating MMP2.