外源性肌醇5磷酸酶对K562细胞增殖及凋亡的影响.docVIP

外源性肌醇5’磷酸酶对K562细胞增殖及凋亡的影响 杨 琳 罗建民 杜行严 杨敬慈 刘晓军 温树鹏 成志勇（河北医科大学第二医院血液科，河北省血液病重点实验室，石家庄 050000） 摘 要：目的通过慢病毒载体介导外源性5’肌醇磷酸酶（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-RT-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±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42.3±7.09，K562/FIV组为77.7±6.33，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SHIP基因抑制细胞增殖促进凋亡。 基因，肌醇5’磷酸酶；慢病毒载体；细胞增殖；细胞凋亡Effect of exogenous SHIP expression on cell growth and apoptosis of K562 cells YANG Lin, LUO Jian-min, DU Xing-yan, YANG Jing-ci, LIU Xiao-jun, WEN Shu-peng, CHENG Zhi-yong.（Province Key Institute of Hematology, Department of Hematology, The Second Hospital of Hebei Midical University, Shijiazhuang 050000, China） Abstract：Objective This study was to investigate the biological function of SHIP gene in cell growth and proliferation. K562 cells by lentiviral vector mediated SHIP. Methods Exogenous SHIP gene was delivered into K562 cells by lentiviral vector-based viral infaction. Expression of SHIP were determined by Western blotting. Transcription of SHIP was examined by FQ-PCR. Changes in cell morphology, cell growth, cell apoptosis after SHIP expression were detected. Results FQ-PCR and Western blot demonstrated that SHIP was successfully expressed in K562 cells. SHIP protein inhibited the growth of K562 cells (inhibition rate=35±3.1 %) and their colony forming capacity ( K562/SHIP group 42.3±7.09 vs K562/FIV group 77.7±6.33, P＜0.05 ). with morphologic features of appoptosis was increased in SHIP-expressing cells and was proved by TUNEL. Conclusions SHIP gene has potential abilities to inhibit leukemia cell proliferation and promote their apoptosis. Key words：ene, SHIP; lentiviral vector; cell proliferation; apoptosis 人类SHIP（SH2 domain containing inositol 5，-phosphatase）基因是继PTEN之后发现的又一肌醇5’磷酸酶，仅限于造血细胞表达[1]。近年来国内外研究表明