Novel Affibody Molecules Targeting the HPV16 E6 Oncoprotein Inhibited the Proliferation of Cervical Cancer Cells.
Novel Affibody Molecules Targeting the HPV16 E6 Oncoprotein Inhibited the Proliferation of Cervical Cancer Cells.
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靶向HPV16 E6癌蛋白的新型脂肪分子抑制了宫颈癌细胞的增殖。
DOI：
10.3389/fcell.2021.677867
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发表时间：
2021
影响因子：
5.5
通讯作者：
Zhang L
中科院分区：
文献类型：
--
作者：
Zhu J;Kamara S;Wang Q;Guo Y;Li Q;Wang L;Chen J;Du Q;Du W;Chen S;Zhu S;Chen J;Chu M;Zhang L
尽管开展了预防性疫苗接种运动，但高危人乳头瘤病毒引起的宫颈癌仍然是妇女的一个重大健康威胁，特别是在发展中国家。这种癌症类型的最初发生和随后的进展主要依赖于E6和E7，这两种组成型表达的关键病毒致癌基因，诱导致癌作用。因此，E6/E7被认为是HPV相关癌症诊断和治疗的理想靶点。本研究从随机噬菌体展示文库中分离了3个新的HPV 16 E6结合抗体分子（ZHPV 16 E61115、ZHPV 16 E61171和ZHPV 16 E61235），并克隆用于细菌生产。通过表面等离子体共振、间接免疫荧光、免疫组织化学和近红外小动物光学成像在体外和体内测定，这些抗体分子显示出对重组和天然HPV 16 E6的高结合亲和力和特异性。此外，ZHPV 16 E61235通过与HPV 16 E6蛋白结合，阻断E6介导的p53降解，从而增加一些关键的p53靶基因的表达，包括BAX、p53 A和p21，从而选择性地降低HPV 16阳性细胞的活力和增殖。重要的是，ZHPV 16 E61235与HPV 16 E7结合体ZHPV 16 E7384组合应用，以同时靶向HPV 16 E6/E7癌蛋白，并且这种组合比单独使用任何一种方式更有效地抑制细胞增殖。机制研究表明，协同抗增殖活性主要取决于诱导细胞凋亡和衰老，而不是细胞周期阻滞。我们的研究结果提供了强有力的证据表明，三种新的HPV 16 E6-结合抗体分子可以形成一个新的基础，为HPV 16阳性癌前病变和肿瘤病变的分子成像和靶向治疗的合理策略的发展。
Despite prophylactic vaccination campaigns, high-risk human papillomavirus (HPV)-induced cervical cancer remains a significant health threat among women, especially in developing countries. The initial occurrence and consequent progression of this cancer type primarily rely on, E6 and E7, two key viral oncogenes expressed constitutively, inducing carcinogenesis. Thus, E6/E7 have been proposed as ideal targets for HPV-related cancer diagnosis and treatment. In this study, three novel HPV16 E6-binding affibody molecules (ZHPV16E61115, ZHPV16E61171, and ZHPV16E61235) were isolated from a randomized phage display library and cloned for bacterial production. These affibody molecules showed high binding affinity and specificity for recombinant and native HPV16 E6 as determined by surface plasmon resonance, indirect immunofluorescence, immunohistochemistry, and near-infrared small animal optical imaging in vitro and in vivo. Moreover, by binding to HPV16 E6 protein, ZHPV16E61235 blocked E6-mediated p53 degradation, which increased the expression of some key p53 target genes, including BAX, PUMA and p21, and thereby selectively reduced the viability and proliferation of HPV16-positive cells. Importantly, ZHPV16E61235 was applied in combination with HPV16 E7-binding affibody ZHPV16E7384 to simultaneously target the HPV16 E6/E7 oncoproteins, and this combination inhibited cell proliferation more potently than either modality alone. Mechanistic studies revealed that the synergistic antiproliferative activity depends primarily on the induction of cell apoptosis and senescence but not cell cycle arrest. Our findings provide strong evidence that three novel HPV16 E6-binding affibody molecules could form a novel basis for the development of rational strategies for molecular imaging and targeted therapy in HPV16-positive preneoplastic and neoplastic lesions.