Scholarlink

Abstract <p>Human papillomavirus (HPV) 18 E6 oncoprotein drives cervical carcinogenesis by degrading p53, enabling uncontrolled cell proliferation. In this study, we developed novel trans-activator of transcription (TAT)-conjugated Affibody molecules (TAT-Z<sub>HPV18E6</sub>) targeting HPV18 E6 for therapeutic applications. High-affinity Affibody variants were screened by phage display using recombinant HPV18 E6 expressed in <i>Escherichia coli</i>. Three candidates (TAT-Z<sub>HPV18E6</sub>: 4, 59, 352) exhibited high binding affinity, with equilibrium dissociation constant (<i>K</i><sub>D</sub>) ranging from 10<sup>−6</sup> to 10<sup>−4</sup>&#xa0;M, with TAT-Z<sub>HPV18E6</sub>: 59 showing superior specificity for native HPV18 E6 in HeLa229 cells, as validated by surface plasmon resonance (SPR), enzyme-linked immunosorbent assay (ELISA), and immuno-fluorescence. In vivo near-infrared fluorescence imaging of DyLight 755-labeled TAT-Z<sub>HPV18E6</sub>: 59 in tumor-bearing mice demonstrated rapid tumor accumulation (peak at 2&#xa0;h) and prolonged retention (&gt; 12&#xa0;h). Mechanistically, TAT-Z<sub>HPV18E6</sub>: 59 restored p53 stability and upregulated pro-apoptotic factors (Bax, PUMA) and cell cycle regulator p21. Notably, combined treatment with TAT-Z<sub>HPV18E6</sub>: 59 and E7-targeting TAT-Z<sub>HPV18E7</sub>: 228 combinatorial enhanced apoptosis and suppressed HeLa229 proliferation, as confirmed by CCK-8 and clonogenic assays. These results demonstrate the utility of TAT-Z<sub>HPV18E6</sub> Affibody molecules as targeted agents, highlighting combinatorial E6/E7 targeting as a potent strategy for HPV18-driven cervical cancer therapy.</p> Key points <p>• <i>TAT-Z</i><sub>HPV18E6</sub>:<i> 59 achieved high affinity for HPV18 E6, enabling precise molecular targeting.</i></p> <p>• <i>Demonstrates dual utility for in vivo imaging and p53-dependent tumor suppression.</i></p> <p>• <i>Dual targeting of E6/E7 yields synergistic therapeutic efficacy in cervical cancer models.</i></p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Human papillomavirus 18 E6-specific Affibody inhibits cervical cancer cell proliferation and induces apoptosis

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年6月26日

查看全文
  • Haiyan Dong,
  • Jing Zhang,
  • Le Wu,
  • Xi Zhang,
  • Jun Chen,
  • Wangqi Du,
  • Saidu Kamara,
  • Shanli Zhu,
  • Lifang Zhang

摘要

Abstract

Human papillomavirus (HPV) 18 E6 oncoprotein drives cervical carcinogenesis by degrading p53, enabling uncontrolled cell proliferation. In this study, we developed novel trans-activator of transcription (TAT)-conjugated Affibody molecules (TAT-ZHPV18E6) targeting HPV18 E6 for therapeutic applications. High-affinity Affibody variants were screened by phage display using recombinant HPV18 E6 expressed in Escherichia coli. Three candidates (TAT-ZHPV18E6: 4, 59, 352) exhibited high binding affinity, with equilibrium dissociation constant (KD) ranging from 10−6 to 10−4 M, with TAT-ZHPV18E6: 59 showing superior specificity for native HPV18 E6 in HeLa229 cells, as validated by surface plasmon resonance (SPR), enzyme-linked immunosorbent assay (ELISA), and immuno-fluorescence. In vivo near-infrared fluorescence imaging of DyLight 755-labeled TAT-ZHPV18E6: 59 in tumor-bearing mice demonstrated rapid tumor accumulation (peak at 2 h) and prolonged retention (> 12 h). Mechanistically, TAT-ZHPV18E6: 59 restored p53 stability and upregulated pro-apoptotic factors (Bax, PUMA) and cell cycle regulator p21. Notably, combined treatment with TAT-ZHPV18E6: 59 and E7-targeting TAT-ZHPV18E7: 228 combinatorial enhanced apoptosis and suppressed HeLa229 proliferation, as confirmed by CCK-8 and clonogenic assays. These results demonstrate the utility of TAT-ZHPV18E6 Affibody molecules as targeted agents, highlighting combinatorial E6/E7 targeting as a potent strategy for HPV18-driven cervical cancer therapy.

Key points

• TAT-ZHPV18E6: 59 achieved high affinity for HPV18 E6, enabling precise molecular targeting.

• Demonstrates dual utility for in vivo imaging and p53-dependent tumor suppression.

• Dual targeting of E6/E7 yields synergistic therapeutic efficacy in cervical cancer models.