抑制核转录因子κBp65对鼻咽癌CNE2细胞的生物学影响

Inhibition of NF-κBp65 expression reduce malignant phenotype of nasopharyngeal carcinoma CNE2 cell

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（siRNA）质粒表达载体（pGPU6/RFP/Neo-NF-κBp65）和阴性对照质粒表达载体（pGPU6/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-κ
Bp65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。

Abstract:

Objective To investigate the biological effects of NF-kappa B p65 (NF-κBp65) on nasopharyngeal carcinoma. Methods NF-κBp65 specific siRNA plasmid expression vector (pGPU6/RFP/Neo-NF-κB 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-κB p65 silencing stable transfected CNE2 cell line (NF-κB p65 silencing group) and negative control cell line (negative control group). The expression of NF-κB p65 gene in cancer cells was analyzed by Western blot. The proliferation and cell cycle of cancer cells were analyzed by MTT 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-κB p65 protein was decreased in cells of NF-κB p65 silencing group. The cell cycle was arrested in S phase and proliferation was reduced after NF-κB p65 was silenced. The growth of transplanted tumor in nude mice of NF-κB 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-κB p65.