CAJ | 학술논문

目的研究蛋白激酶CK2表达沉默对鼻咽癌细胞放射增敏作用,并探讨其可能机制.方法通过RNA干扰技术下调鼻咽癌5-8F细胞中CK2α蛋白表达,利用Western blotting方法验证干扰效果;利用克隆形成实验观察CK2表达沉默后对鼻咽癌细胞放射敏感性的影响;应用免疫荧光方法检测γ-H2AX灶点形成;Annexin-V和PI双染流式细胞仪检测细胞凋亡.结果通过RNA干扰技术可以有效沉默CK2α表达.克隆形成实验结果显示CK2α表达沉默后,鼻咽癌细胞形成克隆能力明显下降,对X线的敏感性明显增加;1 Gy照射后15min,CK2α沉默的细胞γ-H2AX灶点数目明显增加;CK2α沉默的细胞凋亡率明显高于对照组(P<0.01).结论蛋白激酶CK2与鼻咽癌放射敏感性密切相关,CK2可能是一个非常有潜力的鼻咽癌治疗靶点.
목적 연구단백격매CK2표체침묵대비인암세포방사증민작용,병탐토기가능궤제.방법 통과RNA간우기술하조비인암5-8F세포중CK2α단백표체,이용Western blotting방법험증간우효과;이용극륭형성실험관찰CK2표체침묵후대비인암세포방사민감성적영향;응용면역형광방법검측γ-H2AX조점형성;Annexin-V화PI쌍염류식세포의검측세포조망.결과 통과RNA간우기술가이유효침묵CK2α표체.극륭형성실험결과현시CK2α표체침묵후,비인암세포형성극륭능력명현하강,대X선적민감성명현증가;1 Gy조사후15min,CK2α침묵적세포γ-H2AX조점수목명현증가;CK2α침묵적세포조망솔명현고우대조조(P<0.01).결론 단백격매CK2여비인암방사민감성밀절상관,CK2가능시일개비상유잠력적비인암치료파점.
Objective To investigate the effect of protein kinase CK2 gene silencing on the radiosensitization in human nasopharyngeal carcinoma (NPC) cells and its possible mechanism. Methods RNA interference (RNAi) technique was used to down-regulate the protein kinase CK2α expression in 5-8F cells, and clonogenic assay was employed to observe the changes in the radiosensitivity of the cells. DNA double-strand break was assessed by immunofluorescence staining of γ-H2AX foci,and the cell apoptosis was examined using Annexin V-FITC/PI double-staining flow cytometry. Results CK2α protein was successfully silenced by siRNA. CK2α knockdown significantly decreased the clonogenic activity and increased the radiosensitivity of the NPC cells. After a 15-min exposure of the cells to 1 Gy radiation, significant difference occurred in the γ-H2AX foci between CK2α knockdown cells and the control cells (P<0.01). CK2α silencing significantly increased the cell apoptosis after the exposure (P<0.01). Conclusion Protein kinase CK2 plays an important role in the radiosensitivity of the NPC cells, and suppression of its expression might be a potential therapeutic approach of cancer.