HPV16 E6基因变异通过调控BDNF/TrKB表达促进宫颈癌细胞增殖的研究  

HPV16 E6 gene mutation promotes the proliferation of cervical cancer cells by regulating the expression of BDNF/TrKB

作　　者：孙丽艳[1] 杨宾烈[1] 张爱[1] 原杰彦[1] 陶萍萍[1] SUN Li-yan;YANG Bin-lie;ZHANG Ai;YUAN Jie-yan;TAO Ping-ping(Department of Obstetrics and Gynecology, People's Hospital of Shanghai Pudong New Area, Shanghai 201200, China)

机构地区：[1]上海市浦东新区人民医院妇产科,上海201200

出　　处：《海南医学院学报》2021年第4期246-250,共5页Journal of Hainan Medical University

基　　金：上海市浦东新区科学技术发展基金(PKJ2017-Y34)。

摘　　要：目的:明确人乳头瘤病毒E6(HPV16 E6)基因变异对宫颈癌细胞增殖影响的机制研究。方法:采用Real-time PCR检测宫颈癌组织及宫颈上皮内瘤样病变Ⅱ级(CINⅡ)宫颈组织样本中HPV16 E6 T350G、脑源性神经营养因子(BDNF)、酪氨酸激酶受体B(TrKB)及肿瘤蛋白53(p53)mRNA的表达水平;设计并构建HPV16 E6 T350G慢病毒(pLV5-HPV16 E6 T350G)及空载体(pLV5-vector),转染人宫颈上皮细胞(HCerEpiC),Real-time PCR检测HPV16 E6 T350G、BDNF、TrKB及p53 mRNA表达水平,以及Western Blot检测BDNF和TrKB蛋白表达水平及磷脂酰肌醇3激酶/丝/苏氨酸激酶(PI3K/AKT)磷酸化水平;细胞增殖及细胞毒性检测试剂盒(MTT)检测细胞增殖情况。结果:与CINII宫颈组织相比,宫颈癌组织中HPV16 E6 T350G、BDNF、TrKB mRNA表达水平均呈阳性,p53 mRNA表达呈阴性;在HCerEpiC细胞中过表达HPV16 E6 T350G后,可上调BDNF和TrKB蛋白与mRNA表达水平,以及激活BDNF/TrkB下游信号通路PI3K/AKT,并且减少p53蛋白表达水平;HPV16 E6 T350G过表达能够增强HCerEpiC细胞的增殖能力。结论:HPV16 E6 T350G过表达可促进宫颈癌细胞增殖能力,可能与上调BDNF/TrkB表达促进PI3K/AKT信号通路的活化进而降低p53表达有关。Objective:To explore the effect of HPV16 E6 gene mutation on the proliferation of cervical cancer cells.Methods:The expression levels of HPV16 E6 T350G,BDNF,TrKB and p53 mRNA in cervical cancer tissue samples and CINⅡcervical tissues were detected by real-time PCR.HPV16 E6 T350G lentivirus(pLV5-HPV16 E6 T350G)and empty vector(pLV5-vector)were designed and constructed,and transfected with HCerEpiC cells.The expression levels of HPV16 E6 T350G,BDNF,TrKB and p53 mRNA were detected by real-time PCR.Moreover,the expression levels of BDNF,TrKB,PI3K,pPI3K,AKT and pAKT protein were detected by Western blotting analysis.The cell proliferation was evaluated by MTT experiments.Results:Compared with CINⅡcervical tissue,HPV16 E6 T350G,BDNF and TrKB mRNA expression levels were all positive,while p53 mRNA expression was negative.After overexpression of HPV16 E6 T350G in HCerEpiC cells,the expression levels of BDNF and TrKB protein and mRNA were upregulated,and the PI3K/AKT signaling pathway was activated which is the downstream of BDNF/TrKB.Besides,p53 protein expression levels were downregulated.HPV16 E6 T350G overexpression could enhance the proliferation capacity of HCerEpiC cells.Conclusion:Overexpression of HPV16 E6 T350G can promote the proliferation of cervical cancer cells,which may be related to the upregulation of BDNF/TrKB expression,the activation of PI3K/AKT signaling pathway,and the decrease of p53 expression.

关 键 词：HPV16 E6 T350G 宫颈癌 BDNF TRKB 细胞增殖 

分 类 号：R737.33[医药卫生—肿瘤]