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目的探讨易洛魁族同源盒2(iroquois homebox 2,IRX2)基因在骨肉瘤细胞中的表达及其对细胞增殖和细胞周期的影响。方法体外培养骨肉瘤细胞系,应用RT-PCR和Western blot检测各细胞系中IRX2表达;IRX2-siRNA干扰IRX2的表达后,分别用MTT和流式细胞仪检测MG-63细胞的增殖和周期;Western blot检测JAK3和STAT5活化水平。结果 IRX2在骨肉瘤细胞系中的表达显著高于人成骨细胞系(P<0.05);沉默IRX2的表达可显著抑制骨肉瘤细胞系MG-63的增殖,促使其S期细胞比例[(23.93±1.92)%]较对照组[(15.84±1.76)%]增加(P<0.05),而G2/M期细胞比例[(10.44±1.99)%]较对照组[(23.14±3.19)%]降低(P<0.05)。IRX2-siRNA可下调p-JAK3和p-STAT5水平(P<0.05),而过表达IRX2可上调p-JAK3和p-STAT5水平,并促进MG-63细胞增殖(P<0.05)。在JAK3和STAT5抑制剂的作用下,过表达IRX2对MG-63细胞的促增殖作用显著减弱。结论 IRX2可部分通过JAK3/STAT5信号通路调控骨肉瘤细胞的增殖。
Objective To investigate the expression of iroxquois homebox 2 (IRX2) gene in osteosarcoma cells and its effect on cell proliferation and cell cycle. Methods The osteosarcoma cell lines were cultured in vitro. The expression of IRX2 in each cell line was detected by RT-PCR and Western blot. The expression of IRX2 was detected by IRX2-siRNA. The proliferation and the cycle of MG-63 cells were detected by MTT and flow cytometry, respectively. Western blot detection of JAK3 and STAT5 activation levels. Results The expression of IRX2 in osteosarcoma cell lines was significantly higher than that in human osteoblastic cell lines (P <0.05). The silence of IRX2 expression significantly inhibited the proliferation of osteosarcoma cell line MG-63 and promoted the percentage of S phase cells [(23.93 ± 1.92)%] (P <0.05) compared with control group [(15.84 ± 1.76)%], while the proportion of cells in G2 / M phase was (10.44 ± 1.99)% lower than that in control group (23.14 ± 3.19)% (P <0.05). IRX2-siRNA downregulated the expression of p-JAK3 and p-STAT5 (P <0.05), while overexpression of IRX2 upregulated the expression of p-JAK3 and p-STAT5 and promoted the proliferation of MG-63 cells (P <0.05). Under the action of JAK3 and STAT5 inhibitors, the effect of IRX2 overexpression on MG-63 cells was significantly attenuated. Conclusion IRX2 can regulate the proliferation of osteosarcoma cells partly through JAK3 / STAT5 signaling pathway.