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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 effect of ship gene on growth and proliferation of K562 cells by transfecting K562 cells with lentiviral vector-mediated SH2 domain inositol 5’-phosphatase (SHIP) gene. Methods Leukemia cell line K562 was used as a research object to infect K562 cells with lentivirus carrying ship gene. The transcription level of ship was detected by FQ-PCR and the expression of SHIP protein was detected by Western blot. Morphological changes. Results The results of FQ-PCR and Western blot showed that the lentiviral vector pReceiver-Lv31 carrying ship gene could efficiently transfect K562 cells with the positive rate of (74.6 ± 5.8)%. The cell growth curve showed that SHIP could significantly inhibit the growth of K562 cells and the inhibition rate (35.0 ± 3.1)%. Colony formation assay showed that SHIP significantly inhibited the colony-forming ability of K562 cells (36.7 ± 7.1% in K562 / SHIP group and 77.7 ± 6.3% in K562 / FIV group (P < 0.05)]. The apoptosis of cells was observed by morphological observation. SHIP protein was found to promote apoptosis by TUNEL. Conclusion Exogenous expression of ship gene inhibits the proliferation of K562 leukemia cell line and promotes its apoptosis.