过表达SHIP-1对白血病细胞侵袭、迁移及PI3K/AKT信号通路的影响  

Effect of SHIP-1 on Invasion,Migration and PI3K-AKT Signaling Pathway of Leukemic Cells

作　　者：肖喜春[1] 谷月丽[1] 陈淑霞[1] 王根杰[1] 袁林 XIAO Xi-Chun, GU Yue-Li, CHEN Shu-Xia, WANG Gen-Jie, YUAN Lin(Department of Hematology, Shangqiu First People＇s Hospital , Shangqiu 476000, Henan Province, Chin)

机构地区：[1]商丘市第一人民医院血液内科,河南商丘476000

出　　处：《中国实验血液学杂志》2018年第2期324-329,共6页Journal of Experimental Hematology

摘　　要：目的:探讨含有SH2结构的5'肌醇磷酸酶-1(SHIP-1)对白血病细胞增殖、侵袭和迁移能力以及磷脂酰肌醇-3激酶(PI3K)/蛋白激酶B(AKT)信号通路的影响。方法:采用脂质体Lipofectamine 2000将过表达载体p CDNA3.1-SHIP1转染白血病THP-1细胞,实验分为3组:对照组(未做处理的细胞),空载体组(转染空载体p CDNA3.1-NC)和过表达组(转染过表达载体p CDNA3.1-SHIP1);应用CCK-8法检测细胞的增殖情况,Transw ell法检测细胞侵袭、迁移能力,免疫印迹实验(Western blot)检测细胞中SHIP-1、AKT、磷酸化AKT(p AKT)、基质金属蛋白酶-9(MMP-9)蛋白的表达量。结果:细胞转染过表达载体后SHIP-1的表达量显著高于对照组(P<0.05);与对照组相比,空载体组细胞的吸光值没有明显差异(P>0.05),过表达组细胞的吸光值显著降低(P<0.05);空载体组细胞的侵袭、迁移数与对照组无明显差异(P>0.05),过表达组细胞的侵袭、迁移数显著低于对照组(P<0.05);与对照组相比,空载体组细胞中AKT、p AKT和MMP-9的表达量没有显著差异(P>0.05),过表达组细胞中AKT蛋白的表达量无显著差异(P>0.05),但p AKT、MMP-9的表达量均显著降低(P<0.05)。结论:SHIP-1抑制白血病细胞的增殖,且降低其侵袭、迁移能力,其机理可能与通过影响PI3K/AKT信号转导通路的活化从而下调MMP-9的表达有关。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-l, 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.

关 键 词：SHIP-1 白血病 细胞侵袭 细胞迁移 PI3K/AKT信号通路 

分 类 号：R733.7[医药卫生—肿瘤]