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Targeted Biological Effect of An Affitoxin Composed of an HPV16E7 Affibody Fused with Granzyme B (ZHPV16E7-GrB) Against Cervical Cancer In vitro and In vivo

[ Vol. 21 , Issue. 3 ]

Author(s):

Wenhuan Wang, Xiaochun Tan, Jie Jiang, Yiqi Cai, Fangfang Feng, Lifang Zhang* and Wenshu Li*Pages 232-243 (12)

Abstract:


Background: High risk type 16 of human papillomavirus (HPV16) is associated with 50% of cervical cancer, for which reliable targeted therapies are lacking. HPV early protein 7 (E7) is an oncoprotein responsible for cell malignant transformation. In our previous work, a highly specific affibody targeting HPV16E7 (ZHPV16E7) was developed.

Objective: In order to improve the targeted therapeutic effect, the present study prepared an affitoxin consisting of ZHPV16E7 fused with granzyme B (GrB), namely, ZHPV16E7-GrB, and evaluated its targeting action in vitro and in vivo.

Methods: The ZHPV16E7-GrB fusion protein was produced in a prokaryotic expression system. The targeted binding properties of the ZHPV16E7-GrB to the HPV16E7 were confirmed by immunofluorescence assay (IFA) in cervical cancer cell lines, by immunohistochemical assay (IHA) in cervical cancer tissue from clinical specimens and by near-infrared imaging in tumour-bearing mice. The anti- tumour effect on both cervical cancer cells in vitro and tumour-bearing mice in vivo were further evaluated.

Results: A 34-kDa ZHPV16E7-GrB fusion protein was produced in E. coli and displayed the corresponding immunoreactivity. IFA revealed that ZHPV16E7-GrB bound specifically to HPV16-positive TC-1 and SiHa cells. IHA showed that ZHPV16E7-GrB also bound specifically to HPV16-positive clinical tissue specimens. In addition, the near-infrared imaging results showed that ZHPV16E7-GrB was enriched in tumour tissues. Moreover, both the ZHPV16E7-GrB affitoxin and ZHPV16E7 affibody (without GrB) significantly reduced the proliferation of cervical cancer cells in vitro and tumor-bearing mice in vivo, and the anti-proliferative effect of ZHPV16E7-GrB was higher than that of the ZHPV16E7 affibody.

Conclusions: The affitoxin by coupling the affibody with GrB is a promising targeted therapeutic agent with the dual advantages of the targeted affibody and the GrB cytotoxin.

Keywords:

HPV type 16, early protein 7, affibody, granzyme B, cervical cancer, targeted effect.

Affiliation:

Department of Wenzhou Key Laboratory of Gynecology and Obstetrics, Wenzhou People's Hospital, Wenzhou, Zhejiang Province, Department of Microbiology and Immunology, Wenzhou Medical University, Wenzhou, Zhejiang Province, Department of Microbiology and Immunology, Wenzhou Medical University, Wenzhou, Zhejiang Province, Department of Microbiology and Immunology, Wenzhou Medical University, Wenzhou, Zhejiang Province, Department of Microbiology and Immunology, Wenzhou Medical University, Wenzhou, Zhejiang Province, Department of Microbiology and Immunology, Wenzhou Medical University, Wenzhou, Zhejiang Province, Department of Microbiology and Immunology, Wenzhou Medical University, Wenzhou, Zhejiang Province

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