PNCK对鼻咽癌细胞增殖和凋亡作用的研究

目的 研究妊娠上调的非泛素性钙调蛋白激酶(Pregnancy-upregulated nonubiquitous calmodulin kinase,PNCK)在鼻咽癌组织中的表达以及对鼻咽癌细胞增殖和凋亡的作用。方法 采用免疫组织化学方法检测鼻咽癌癌组织和正常组织中PNCK表达水平。构建慢病毒载体干扰PNCK基因表达,并采用Real time PCR和免疫印迹结果证实,采用MTT方法、流式细胞术、Caspase3和Caspase7活性检测鼻咽癌肿瘤细胞增殖和凋亡的变化。结果 免疫组织化学结果显示PNCK蛋白在鼻咽癌癌组织中呈特异性高表达。Real time PCR和免疫印迹结果证实慢病毒成功干扰鼻咽癌CNE-2Z细胞中PNCK表达(PNCK基因在mRNA和蛋白水平表达受到显著抑制)。MTT检测结果显示抑制PNCK表达抑制CNE-2Z细胞增殖。此外,干扰PNCK表达引起CNE-2Z细胞Caspase3和Caspase7活性上升,流式细胞术表明干扰PNCK基因表达可以促进CNE-2Z细胞凋亡。结论 干扰PNCK基因表达抑制鼻咽癌细胞增殖并诱导肿瘤细胞凋亡,可能是治疗鼻咽癌的潜在靶点。

Effects of PNCK on proliferation and apoptosis of nasopharyngeal carcinoma cells

Objective The aim of this study was to explore the expression of pregnancy-upregulated nonubiquitous calmodulin kinase(PNCK)in nasopharyngeal carcinoma(NPC)and its effect on proliferation and apoptosis of NPC cells.Methods Immunohistochemistry was used to detect the expression of PNCK in NPC and normal tissues.The lentiviral vector was constructed to interfere with the expression of PNCK gene,confirming by real-time PCR and Western blot assays.The MTT assay,Annexin V staining with flow cytometry(FCM)analysis and caspase 3/7 activity assays were used to detect proliferation and apoptosis of CNE-2Z cells knocked-down PNCK.Results PNCK protein was highly expressed in NPC tissues.RT-qPCR and Western blot results confirmed that lentivirus successfully interfered with PNCK expression in nasopharyngeal carcinoma CNE-2Z cells.MTT assay result confirmed that the knock-down of PNCK significantly inhibited proliferation and induced apoptosis of CNE-2Z cells.Conclusion Interfering with PNCK gene expression inhibits proliferation of nasopharyngeal carcinoma cells and induces cell apoptosis,which can be potential targets for cancer treatment of nasopharyngeal carcinoma.