反义NF-κB p65寡核苷酸对食管鳞癌细胞EC9706增殖及凋亡的影响  

Effects of antisense NF-κB p65 oligonucleotide on the proliferation and apoptosis in esophageal squamous carcinoma cell line EC9706

作　　者：轩小燕[1,2] 王新华[1] 李珊珊[1] 张红燕[1] 王小军[1] 李娜[1] 王丰[1] 阎爱华[1] 

机构地区：[1]郑州大学第一附属医院病理科 [2]河南省肿瘤病理重点实验室,郑州450052 [2]郑州大学基础医学院微生物学与免疫学教研室,郑州450001

出　　处：《郑州大学学报（医学版）》2009年第3期472-475,共4页Journal of Zhengzhou University(Medical Sciences)

基　　金：河南省科技攻关基金资助项目0624410048;072102310042

摘　　要：目的:探讨反义NF-κBp65寡核苷酸对食管鳞癌细胞EC9706增殖、凋亡及细胞周期的影响。方法:利用转染试剂KeyGenI介导将反义NF-κBp65寡核苷酸片段体外转染入人食管鳞癌EC9706细胞株。转染72h后,应用RT-PCR、流式细胞术检测EC9706细胞中NF-κBp65mRNA和蛋白的表达;MTT法检测细胞增殖能力;流式细胞术检测细胞凋亡和细胞增殖周期。以单纯转染试剂为阴性对照,未转染的EC9706细胞为空白对照。结果:转染72h后,反义组EC9706细胞中NF-κBp65mRNA的表达及蛋白标记率与阴性对照和空白对照组比较,下降明显,差异具有统计学意义(P<0.05);反义组EC9706细胞的增殖能力与阴性对照和空白对照组相比明显受抑,抑制率为15.0%;反义组EC9706凋亡细胞数较阴性对照、空白对照组升高,差异具有统计学意义(P<0.05);反义组G1期的EC9706细胞数较阴性对照、空白对照组升高,而S期细胞数较阴性对照、空白对照组降低,差异均具有统计学意义(P均<0.05)。结论:特异性阻断NF-κBp65的转录,可以抑制肿瘤细胞增殖,促进肿瘤细胞凋亡,参与细胞周期调控。NF-κBp65有望成为肿瘤基因治疗的靶点。Aim ： To investigate the effects of antisense NF-κB p65 oligonucleotides on proliferation,apoptosis and cell eycle in esophageal squamous carcinoma cell line EC9706. Methods：Using KeyGenl-mediated transfeetion, the antisense NF-κB p65 oligonucleotides were transfeeted into EC9706 cell line. The expression of NF-κB p65 was confirmed by RT- PCR and flow cytomertry respectively. MTT assay was used to detect cell proliferation ability and flow cytomertry was used to detect the cell apoptosis and cell cycle. EC9706 Cells transfected with KeyGenl as negative control and EC9706 cells as blank control. Results： 72 h after transfection , the NF-κB p65 mRNA and protein expression in EC9706 cells transfected with antisense NF-κB p65 oligonucleotides were obviously lower than those in negative control and blank control groups （ P 〈 0.05 ） ; the proliferation ability in EC9706 cells transfected with antisense NF-κB p65 oligonucleotides was inhibited more than those in control groups, and the inhibition rate was 15.0% ; In EC9706 cells transfected with antisense NF-κB p65 oligonucleotides, the apoptosis rate was obviously higher than those in control groups（ P 〈 0.05 ）and had more G, phase cells ,fewer S pbase cells than those in control groups（ P 〈 0.05 ）. Conclusion ： Transfection of antisense NF-κB p65 oligo- nucleotides leads to suppression of proliferation, induction of G1 arrest and apoptosis in EC9706 cells and NF-κB p65 has the potential to be a new target in cancer gene therapy.

关 键 词：NF-κB P65基因 反义寡核苷酸技术 MTT 流式细胞术 

分 类 号：R735.1[医药卫生—肿瘤]