Kv1.3钾通道在骨肉瘤中的表达及作用研究

目的 探讨特异性短发卡RNA（shRNA）抑制Kv1.3钾通道对骨肉瘤MG-63细胞增殖和凋亡的影响。方法 采用实时荧光定量聚合酶链反应、Western blotting和免疫组化检测骨肉瘤MG-63细胞中Kv1.3钾通道的表达，并分别采用CCK-8法、裸鼠骨肉瘤异种移植模型和流式细胞仪检测MG-63细胞增殖、生长和凋亡的变化，采用Western blotting检测MG-63细胞中多聚ADP核糖聚合酶（PARP）和caspase-3的表达水平。结果Kv1.3钾通道在骨肉瘤细胞中异常高表达。沉默Kv1.3钾通道的Ad5-Kv1.3-shRNA能从体内外有效地抑制MG-63细胞增殖并促进其早期凋亡。沉默Kv1.3钾通道可明显诱导细胞凋亡相关蛋白caspase-3和PARP的裂解。结论Kv1.3钾通道参与骨肉瘤细胞增殖和凋亡的过程，有望成为骨肉瘤治疗及诊断的新靶点基因。

Expression and role of Kv1.3 potassium channel in human osteosarcoma

Objective To explore the effects of silencing Kv1.3 potassium channel by short hairpin RNA (shRNA) on the proliferation and apoptosis of osteosarcoma cells.Methods The expression of Kv1.3 potassium channel in osteosarcoma MG-63 cells was detected by real time quantitative polymerase chain reaction, Western blotting and immunohistochemistry.Next, we measured the proliferation, growth and cell apoptosis of MG-63 cells using CCK-8, xenograft model in vivo and flow cytometry.Finally, Western blotting was performed to detect the expression of cell apoptosis related genes including poly ADP-ribose polymerase (PARP) and caspase-3 in MG-63 cells.Results Kv1.3 potassium channel was overexpressed in osteosarcoma cells and tissues.Knockdown of Kv1.3 by shRNA significantly suppressed osteosarcoma cell proliferation and inhibited the growth of MG-63 in vivo and in vitro.Additionally, cell early apoptosis was also induced.Silencing Kv1.3 gene enhanced the leavel of cleaved caspase-3 and cleaved PARP.Conclusion Kv1.3 potassium channel is involved in proliferation and apoptosis process of human osteosarcoma and may represent a potential target for osteosarcoma diagnosis and treatment.