外源性肌醇5′磷酸酶对K562细胞增殖及凋亡的影响

Effects of exogenous SHIP expression on cell growth and apoptosis of K562 cells

目的通过慢病毒载体介导外源性5′肌醇磷酸酶(SH2domain contaihing inositol5′-phosphatase,SHIP)基因转染K562细胞,探讨ship基因对K562细胞生长增殖的影响。方法以白血病细胞株K562为研究对象,将携带ship基因的慢病毒感染K562细胞,FQ-PCR方法检测ship转录水平,Western blot方法检测转染后SHIP蛋白表达变化;比较ship基因表达前后细胞增殖、形态的变化。结果FQ-PCR和Western blot结果显示,携带ship基因的慢病毒载体pReceiver-Lv31能高效转染K562细胞,阳性率(74.6±5.8)%;细胞生长曲线显示SHIP可显著抑制K562细胞生长,抑制率(35.0±3.1)%;集落形成实验显示SHIP能显著抑制K562细胞集落形成能力[K562/SHIP组集落形成率(36.7±7.1)%,K562/FIV组为(77.7±6.3)%,(P<0.05)];细胞形态观察发现凋亡增加,TUNEL法证实SHIP蛋白促进细胞凋亡。结论外源性ship基因表达抑制白血病细胞系K562的增殖活性,并促进其凋亡。

Objective To investigate the biological function of SH2 domain containing inositol 5＇ -phos- phatase （ship） gene in cell growth and proliferation. K562 ceils by lentiviral vector mediated ship. Methods Exogenous ship gene was delivered into K562 cells by lentiviral vector-based viral infection. SHIP expression was determined by Western blot. Transcription of ship was examined by FQ-PCR. Changes in cell morphology, cell growth, cell apoptosis after SHIP expression were observed. Results FQ-PCR and Western blot demonstrated successful SHIP expression in K562 cells. SHIP protein inhibited the growth of K562 cells [ inhibition rate = （35.0 ± 3.1 ） % ] and their colony forming capacity [ K562/SHIP group （36.7 ± 7.1 ） % vs K562/FIV group （77.7 ± 6.3 ） % , P 〈 0. 05 ]. Morphological observation showed enhanced cell apoptosis. TUNEL confirmed the enhanced cell apoptosis by SHIP protein. Gonclusion ship gene has the potential ability to inhibit leukemic cell proliferation but enhance cell apoptosis.