PMID: 22575191May 12, 2012Paper

Effect of SHIP mutation on invasion and migration of K562 leukemia cells

Zhonghua xue ye xue za zhi = Zhonghua xueyexue zazhi
Xiao-Jun LiuJian-Min Luo

Abstract

To explore the effect of mutation in PxxP domain of SHIP on migration and invasion of leukemia cells and its mechanism. The lentiviral vector mediated wild type SHIP (wtSHIP) and mutant SHIP (muSHIP) plasmids were transfected into K562 cells through gene transfection techniques. Expression of SHIP at mRNA and protein level was detected by real-time PCR and Western blot, respectively. Transwell assay was used to analyze the difference between the migration and invasion ability of the K562/wtSHIP and the K562/muSHIP cells after transfection. Primary migration associated factor FAK, MMP and NF-κB were assayed by Western blot. After transfection, the SHIP expression in transfected K562 cells were significantly increased. Compared with the migration ability of K562/wtSHIP\[(15.8 ± 1.4)%\], that of K562/muSHIP cells \[(54.3 ± 2.4)% \] increased greatly and almost at the same level of that of K562/pFIV\[(50.3 ± 3.8)%\] (P < 0.01). The invasion assay also showed that K562/wtSHIP\[(32 ± 6)/HP\] has a lower invasion ability than that of the K562/muSHIP group \[(83 ± 16)/HP\] and K562/pFIV group \[(78 ± 13)/HP\] (P < 0.01). Western blot analysis showed that the expression of p-FAK and NF-κB was up-regulated in K562/muSHIP group compared t...Continue Reading

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