抑制核转录因子κBp65对鼻咽癌CNE2细胞的生物学影响

发布时间：2018-01-24 01:29

本文关键词： 鼻咽癌核转录因子-κ Bp 小干扰核糖核酸　出处：《中国现代医学杂志》2017年08期 　论文类型：期刊论文

【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,P0.05)。结论 NF-κBp65沉默能降低鼻咽癌CNE2细胞的恶性生物学行为。
[Abstract]:Objective to investigate the biological effect of nuclear factor 魏 B p65 (NF- 魏 Bp65) silencing on nasopharyngeal carcinoma (NPC). Methods to construct NF- 魏 Bp65 specific small interfering RNA siRNAs (siRNAs). Plasmid expression vector pGPU 6% RFP / Neo-NF- 魏 Bp65) and negative control plasmid vector pGPU 6% RFP / Neo-NC). The recombinant plasmid expression vector was transfected into human nasopharyngeal carcinoma (NPC) CNE2 cells with non-liposome lipids. To establish the stable transfection of NF- 魏 Bp65 into CNE2 cell line (NF- 魏 Bp65 silencing group) and negative control cell line (negative control group); The expression of NF- 魏 Bp65 gene in cancer cells was analyzed by Western blot. MTT assay and flow cytometry were used to analyze the proliferation ability and cell cycle change of cancer cells. The effect of silencing NF- 魏 Bp65 on tumorigenicity of cancer cells was analyzed in nude mice. Results stable transfection of CNE2 cells was successfully achieved. The expression of NF- 魏 Bp65 protein was decreased in NF- 魏 Bp65 silencing group. After the silencing of NF- 魏 Bp65, the cell cycle of CNE2 was arrested in S phase, and the proliferation rate was slowed down. Compared with negative control group and blank control group, the growth of xenograft tumor in NF- 魏 Bp65 silencing group was slow and light, the difference was statistically significant (P < 0.05). Conclusion NF- 魏 Bp65 silencing can reduce the malignant biological behavior of nasopharyngeal carcinoma CNE2 cells.
【作者单位】：桂林医学院科学实验中心;桂林医学院附属医院;
【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)
【分类号】：R739.63
【正文快照】： 鼻咽癌起源于鼻咽黏膜上皮,在我国南方和东南亚地区高发,由于其发病部位隐匿,早期症状不明显,确诊时多数患者已达中晚期,因此,治疗效果差,5年生存率低,主要原因是局部复发和远处转移[1]。核转录因子-κB(nuclear transcription factor-κB,NF-κB)是一个核转录因子家族,静息状

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