人乳头状瘤病毒感染宫颈病变程度与E2、E6、E7基因表达水平相关性研究  

Correlation between the degree of HPV-infected cervical lesions and the expression levels of E2, E6 and E7 genes

作　　者：辛玲丽 程波 孙锁柱 熊一诺 陈玲 丁晓萍 侯庆香 XIN Ling-li;CHENG Bo;SUN Suo-zhu;XIONG Yi-nuo;CHEN Ling;DING Xiao-ping;HOU Qing-xiang(Department of Obstetrics and Gynecology,PLA Rocket Force Medical Center,Beijing100088,China;Department of Pathology,PLA Rocket Force Medical Center,Beijing100088,China;Jining Medical University,Jining272013,China)

机构地区：[1]火箭军特色医学中心妇产科,北京100088 [2]火箭军特色医学中心病理科,北京100088 [3]济宁医学院,山东济宁272013

出　　处：《诊断病理学杂志》2021年第12期1040-1043,1047,共5页Chinese Journal of Diagnostic Pathology

摘　　要：目的从分子水平探讨E2及E6、E7基因在子宫颈上皮内瘤变(CIN)及宫颈癌发病过程中的相关性及临床意义。方法纳入2018-02—2019-10期间感染HPV16型的因CIN及宫颈癌就诊于我院行宫颈活检术、宫颈锥切术、宫颈癌根治术的患者100例为实验组,同期因其他子宫良性肿瘤于我院行全子宫切除术的患者15例作为对照组,采用RT-PCR检测方法,对两组患者液基细胞标本行HPV E2、E6、E7基因的检测。分别以HPV16 E2基因的原核表达质粒pcDNA3.1-HPV16 E2及pcDNA3.1空载质粒转染SiHa细胞,检测其E2、E6、E7基因表达水平。结果对照组患者未检出E2、E6、E7基因的表达;实验组患者均检出E2、E6、E7基因的表达,且在CINⅠ、CINⅡ、CINⅢ及宫颈癌患者的液基细胞中,HPV E2、E6及E7基因检出率的差异存在统计学意义(P<0.05),随着CIN级别的增加,HPV E2检出率呈下降趋势,E6、E7基因检出率呈上升趋势。SiHa细胞过表达E2基因后,E2基因表达量上升37.42倍,而E6基因表达量为对照组的0.28倍,E7基因表达量为对照组的0.27倍。结论 E2、E6、E7基因与CIN、宫颈癌病程发展密切相关,E2基因负向调控E6、E7基因,可以作为临床上常规宫颈癌筛查方法以外的补充检测方法。Objective To investigate the correlation and clinical significance of E2, E6 and E7 genes in the pathogenesis of cervical intraepithelial neoplasia(CIN) and cervical cancer at the molecular level. Methods By using real-time fluorescent quantitative PCR(RT-PCR), 100 cases of CIN and cervical cancer patients with HPV16 infection were enrolled as experimental group, and 15 patients underwent hysterectomy in our hospital due to other benign uterine tumors in the same period were included as control group. The HPV E2, E6 and E7 genes were detected in liquid-based cell samples. The prokaryotic expression plasmid pcDNA3.1-HPV16 E2 was constructed, and SiHa cells were transfected with pcDNA3.1-HPV16 E2 recombinant plasmid(experimental group) or pcDNA3.1 empty plasmid(control group). The mRNA expression levels of E2, E6 and E7 genes were evaluated. Results The expression of E2, E6 and E7 genes was found in patients of the experimental group, rather than in patients of the control group, and the incidences of HPV E2, E6 and E7 gene expression were statistically different among CIN I, CIN II, CIN III and cervical cancer patients(P<0.05). Along with the increase of CIN grade, HPV E2 expression showed a downward trend, while E6 and E7 gene expression showed upward trends. In SiHa cells transfected with HPV16 E2 recombinant plasmid, the expression of E2 gene increased by 37.42 times, while the expression of E6 and E7 gene was 0.28 and 0.27 times of those in the control group, respectively. Conclusion The E2, E6, and E7 genes are closely related to the grade of CIN and cervical cancer. E2 negatively regulates the E6 and E7 genes expression. Expression of E2, E6 and E7 can be used as supplementary biomarkers for routine cervical cancer screening.

关 键 词：HPV16 宫颈癌 宫颈上皮内瘤变 E2基因 E6基因 E7基因 

分 类 号：R73[医药卫生—肿瘤]