中文摘要：

目的：探讨SHIP基因对人白血病细胞系K562细胞周期的调控作用。方法：用携带野生型SHIP基因及绿色荧光蛋白（green fluorescent protein,GFP）基因的慢病毒及空载体慢病毒转染人白血病K562细胞。FCM检测转染效率、细胞凋亡率及细胞周期变化,实时荧光定量PCR（real-time fluorescent quantitative PCR,RFQ-PCR）检测SHIPmRNA水平的变化,Western印迹法检测SHIP、cyclinD1、p21^WAF1/CIPI、p27^KIP1蛋白表达水平的变化。结果：与转染空载体组和未转染组相比,转染野生型SHIP基因的K562细胞增殖减慢,凋亡率明显上升（P〈0.05）;细胞周期显示,G0/G1期延长,G1期前出现亚二倍体的凋亡峰,S期和G2/M期比例降低;cyclin D1表达降低,p21^WAF1/CIPI和p27^KIP1表达增加。结论：转染野生型SHIP基因可使细胞周期阻滞在G0/G1期,抑制白血病细胞增殖,促进白血病细胞凋亡。

英文摘要：

Objective：To investigate the regulating effect of SHIP gene on the cell cycle of human leukemia cell line K562 in vitro. Methods： The recombinant plasmid harboring FIV-SHIP and green fluorescent protein （GFP） were transfected into human leukemia K562 cells. The K562 cells transfected with and without empty vector were as control groups. Flow cytometry was used to detect the transfection efficiency, apoptotic rate, and cell cycle changes. The mRNA level of SHIP were measured by using real-time fluorescent quantitative PCR （FQ-PCR）, and the protein levels of SHIP, cyclin D1, p21^WAF1/CIPI, and p27^KIP1 were determined by Western blotting. Results： As compared with that in control groups, the growth of K562 cells transfected with wild type of SHIP gene was slowed down and the apoptotic rate was significantly increased（P〈0.05）. Flow cytometry analysis showed that the K562/SHIP cells were arrested in G0/G1 phase and displayed sub-diploid apoptosis peak and the proportion of cells in S and G2/M phase was decreased. The expression of cyclin D1 was decreased and the expression of p27^KIP1 and p21^WAF1/CIPI was increased. Conclusion： Transfection of wild type of SHIP gene arrested K562 cells in G0/G1 phase, inhibits the proliferation, and induces the apoptosis in vitro.