目的:观察干扰素(interferon-α2b;IFN-α2b)对HEL细胞(人红白血病细胞株)生长、凋亡及JAK2 V617F突变基因表达的影响。方法:HEL细胞置于含10%胎牛血清的RPMI1640培养基中培养,干扰素浓度分别为0、5×105、10×105、50×105、100×105 U/L 5个实验组,收集各组不同培养时间(0、24、72、120 h)的细胞在倒置光学显微镜下观察细胞的生长状况;MTT法检测干扰素对HEL细胞增殖的抑制作用,采用流式细胞术检测细胞凋亡。应用荧光定量PCR检测JAK2 V617F突变基因的表达水平。结果:干扰素在5×105 U/L、10×105 U/L、50×105 U/L、100×105 U/L 对 HEL 细胞增殖的抑制率分别是18.57%、25.10%、42.10%、57.00%。干扰素在100×105 U/L 浓度作用24、72、120 h JAK2 V617F/GAPDH 荧光定量值分别为1.556、1.213、0.870。结论:干扰素α2b能抑制HEL细胞增殖、诱导HEL细胞凋亡,随干扰素浓度增加HEL细胞凋亡率增加,干扰素能抑制HEL细胞JAK2 V617F突变基因的表达,其作用呈剂量依赖性。
Objective To observe the effect of Interferon-α2b on HEL cells (human erythroleukemia cell line) growth, apoptosis and JAK2 V617F mutation gene expression. Methods HEL cells were placed in RPMI1640 containing 10% FBS and incubated in a cell incubator. Cells in the logarithmic growth phasem were collected, adjusting the cell density to 1 × 105/mL for experimental research. The interferon concentration in five groups were 0, 5 × 105, 10 × 105, 50 × 105, 100 × 105 U/L, with different incubation time (0, 24, 72, 120 h), respectively. The cell growth status in different groups was observed in the inverted optical microscope; MTT was used to detect the inhibition of interferon on HEL cell proliferation. Cell apoptosis was detected by flow cytometry. Fluorescence quantitative PCR was used to detect the mutation gene of JAK2 V617F expression. Results Inhibition rates of Interferon on the HEL cell proliferation in 5 × 105 U/L, 10 × 105 U/L, 50 × 105 U/L, 100 × 105 U/L groups were 18.57%, 25.10%, 42.10%, 57.00%, respectively. JAK2 V617F/GAPDH by fluorescence quantitative was 1.556, 1.213, 0.870 respectively under the concentration of interferon 100 × 105 U/L for 24, 72, 120 h. Conclusions Interferon-α2b can inhibit HEL cells proliferation and induce HEL cells apoptosis. Increasing concentration of interferon increases HEL cell apoptosis rate. Interferon can inhibit JAK2 V617F expression of HEL cells in a dose-dependent manner.