摘要
目的探讨核转录因子κBp65(NF-κBp65)沉默对鼻咽癌的生物学作用。方法构建NF-κBp65特异性小干扰核糖核酸(si RNA)质粒表达载体(p GPU6/RFP/Neo-NF-κBp65)和阴性对照质粒表达载体(p GPU6/RFP/Neo-NC)。采用非脂质体脂类转染剂将重组质粒表达载体转入人鼻咽癌CNE2细胞,建立NF-κBp65沉默的稳定转染CNE2细胞株(NF-κBp65沉默组)和阴性对照细胞株(阴性对照组);蛋白质印迹法(Western blot)分析癌细胞中NF-κBp65基因的表达水平,MTT实验及流式细胞术分析癌细胞的增殖能力和细胞周期变化;复制裸鼠移植瘤模型,分析沉默NF-κBp65对癌细胞成瘤性的影响。结果成功获得稳定转染CNE2细胞株,NF-κBp65沉默组细胞中NF-κBp65蛋白表达水平降低;NF-κBp65沉默后,CNE2细胞周期阻滞于S期,增殖速度减慢;NF-κBp65沉默组裸鼠移植瘤生长慢,重量轻,与阴性对照组和空白对照组比较,差异有统计学意义(F=14.386,P<0.05)。结论 NF-κBp65沉默能降低鼻咽癌CNE2细胞的恶性生物学行为。
Objective To investigate the biological effects of NF-kappa B p65 （NF-KBp65） on nasopha- ryngeal carcinoma. Methods NF-KBp65 specific siRNA plasmid expression vector （pGPU6/RFP/Neo-NF-KB p65） and negative control plasmid expression vector （pGPU6/RFP/Neo-NC） were constructed and transfected into human nasopharyngeal carcinoma CNE2 cell line by non-liposomal lipid transfection agent to establish NF-KB p65 silencing stable transfected CNE2 cell line （NF-KB p65 silencing group） and negative control cell line （negative control group）. The expression of NF-KB p65 gene in cancer cells was analyzed by Western blot. The proliferation and cell cycle of cancer cells were analyzed by MTI＂ assay and Flow Cytometry. A xenograft model in nude mice was established to analyze the tumor igenicity of cancer cells. Results The stable transfected CNE2 cell lines were successfully obtained. The expression of NF-KB p65 protein was decreased in cells of NF-KB p65 silencing group. The cell cycle was arrested in S phase and proliferation was reduced after NF-KB p65 was silenced. The growth of transplanted tumor in nude mice of NF-KB p65 silencing group was slower, and weight of transplanted tumor was lighter than that in negative control group or blank control group, the difference was statistically significant （F= 14.386, P〈 0.05）. Conclusions Malignant biological phenotype of nasopharyngeal carcinoma CNE2 cell line could be reduced by silencing NF-KB p65.
出处
《中国现代医学杂志》
CAS
北大核心
2017年第8期7-11,共5页
China Journal of Modern Medicine
基金
国家自然科学基金资助课题(No:81260344)
广西自然科学基金资助课题(No:2013GXNSFAA019228)
广西壮族自治区教育厅资助课题(No:LX2014274)
