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[Effect of SHIP-1 on Invasion, Migration and PI3K-AKT Signaling Pathway of Leukemic Cells].
Zhongguo Shi Yan Xue Ye Xue za Zhi 2018 April
OBJECTIVE: To investigate the effect of SH2-containing inositol phosphatase-1 (SHIP-1) on the proliferation, invasion and migration of human leukemia cells as well as phosphatidylinositol-3 kinase (PI3K) / protein kinase B (AKT) signaling pathway.
METHODS: The overexpression vector pCDNA3.1-SHIP1 was transfected into THP-1 cells by Lipofectamine 2000. The experiment was divided into 3 groups: control group (untreated cells) and empty vector group (transfected with empty vector pCDNA3.1-NC) and overexpression group (transfected with overexpression vector pCDNA3.1-SHIP1). The cell proliferation was tected by CCK-8 assay, Transwell assay was used to evaluate the cell invasion and migration capabilities. The expressions of SHIP-1, AKT, phosphorylated AKT (pAKT), matrix metalloproteinase-9 (MMP-9) protein were analyzed by Western blot.
RESULTS: The expression of SHIP-1 in overexpression group was significantly higher than that in the control group(P<0.05). Compared with the control group, the absorbance of the cells in the empty vector group was not statistically different (P>0.05), and the absorbance in overexpression group decreased significantly(P<0.05). The cell numbers of invasion and migration were not significantly different between empty and control groups(P>0.05), but those in overexpression group were significantly lower than those in the control group(P<0.05). Compared with the control group, the expression of AKT, pAKT and MMP-9 in the empty vector group was not statistically different (P>0.05); the AKT protein in overexpression group was not significantly different (P>0.05), but the pAKT and MMP-9 significantly decreased(P<0.05).
CONCLUSION: SHIP-1 plays a role in inhibiting the proliferation, invasion and migration of leukemia cells, the mechanism probably relates with supressing the expression of MMP-9 by regulating PI3K/AKT signaling pathway.
METHODS: The overexpression vector pCDNA3.1-SHIP1 was transfected into THP-1 cells by Lipofectamine 2000. The experiment was divided into 3 groups: control group (untreated cells) and empty vector group (transfected with empty vector pCDNA3.1-NC) and overexpression group (transfected with overexpression vector pCDNA3.1-SHIP1). The cell proliferation was tected by CCK-8 assay, Transwell assay was used to evaluate the cell invasion and migration capabilities. The expressions of SHIP-1, AKT, phosphorylated AKT (pAKT), matrix metalloproteinase-9 (MMP-9) protein were analyzed by Western blot.
RESULTS: The expression of SHIP-1 in overexpression group was significantly higher than that in the control group(P<0.05). Compared with the control group, the absorbance of the cells in the empty vector group was not statistically different (P>0.05), and the absorbance in overexpression group decreased significantly(P<0.05). The cell numbers of invasion and migration were not significantly different between empty and control groups(P>0.05), but those in overexpression group were significantly lower than those in the control group(P<0.05). Compared with the control group, the expression of AKT, pAKT and MMP-9 in the empty vector group was not statistically different (P>0.05); the AKT protein in overexpression group was not significantly different (P>0.05), but the pAKT and MMP-9 significantly decreased(P<0.05).
CONCLUSION: SHIP-1 plays a role in inhibiting the proliferation, invasion and migration of leukemia cells, the mechanism probably relates with supressing the expression of MMP-9 by regulating PI3K/AKT signaling pathway.
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