Zhu Jiang, Chen Lingjing, Zheng Qiaoli, Han Rui, Chen Xianzhen, Zhou Qiang, Cheng Hao
Department of Dermatology, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou 310016, Zhejiang Province, China.
Department of Dermatology, Hangzhou Children's Hospital, Hangzhou 310014, Zhejiang Province, China.
J Cancer. 2021 Oct 20;12(23):7130-7137. doi: 10.7150/jca.60554. eCollection 2021.
Almost all cervical cancer is associated with persistent infection of high-risk (hr) human papillomavirus (HPV) like HPV16. The E7 oncoprotein encoded by hrHPV plays a crucial role in carcinogenesis. The present study aimed to establish a reliable protocol of immunohistochemistry stains to detect HPV16 E7 protein in formalin-fixed and paraffin-embedded tissue specimens of various cervical lesions. Firstly, the HPV16 E7 gene was inserted into a prokaryotic expression vector pGEX-4T2. Then the recombinant plasmid pGEX-4T2-(HPV16-E7) was transformed into JM109. The fusion protein containing a GST tag was purified, and New Zealand white rabbits were immunized to produce the HPV16 E7 polyclonal antibody. The anti-HPV16 E7 antibody was verified by western blotting and immunofluorescence stains, and applied in 182 HPV16 DNA-positive cervical tissue specimens and matched 36 HPV DNA-negative specimens by immunohistochemistry. Furthermore, A positive correlation between HPV16 E7 protein expression and malignancy grade was observed. But there is no relationship between HPV16 E7 protein expression and tumor sizes, tumor differentiation, lymph node metastasis, International Federation of Gynecology and Obstetrics (FIGO) stage, or lymphovascular space invasion in cervical cancer. These findings provide a basis for further research focusing on the role of HPV E7 protein in various HPV-related diseases.
几乎所有宫颈癌都与高危（hr）人乳头瘤病毒（HPV）如HPV16的持续感染有关。hrHPV编码的E7癌蛋白在致癌过程中起关键作用。本研究旨在建立一种可靠的免疫组织化学染色方案，以检测各种宫颈病变的福尔马林固定石蜡包埋组织标本中的HPV16 E7蛋白。首先，将HPV16 E7基因插入原核表达载体pGEX-4T2。然后将重组质粒pGEX-4T2-(HPV16-E7)转化到JM109中。纯化含有GST标签的融合蛋白，并用其免疫新西兰白兔以产生HPV16 E7多克隆抗体。通过蛋白质免疫印迹法和免疫荧光染色对该抗HPV16 E7抗体进行验证，并通过免疫组织化学法将其应用于182例HPV16 DNA阳性宫颈组织标本及与之匹配的36例HPV DNA阴性标本。此外，观察到HPV16 E7蛋白表达与恶性程度之间呈正相关。但在宫颈癌中，HPV16 E7蛋白表达与肿瘤大小、肿瘤分化、淋巴结转移、国际妇产科联盟（FIGO）分期或脉管间隙浸润均无关。这些发现为进一步研究HPV E7蛋白在各种HPV相关疾病中的作用提供了依据。
