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<p>Cervical cancer is a significant cause of cancer-related mortality in women, and the toxicity of conventional anticancer drugs has necessitated the development of novel therapeutic strategies. In the present study, we designed a fusion protein containing CpsA and CpsC proteins encoding capsular polysaccharides in <i>Streptococcus agalactiae</i> and evaluated its potential anticancer activity. Physicochemical properties, 3-dimensional (3D) structure, and molecular dynamics (MD) simulation were analyzed using in silico tools. Then, the recombinant gene was cloned, expressed in a bacterial host, and purified successfully. Finally, the anticancer effects of the CpsA-CpsC fusion protein on cervical cancer cell line (HeLa) were evaluated through MTT, Real-Time PCR, and flow cytometry techniques. Functional assays demonstrated that the CpsA–CpsC fusion protein exerted time- and dose-dependent cytotoxic effects on HeLa cells, with an IC50 value of 68.1&#xa0;µg/mL at 48 after 48&#xa0;h of treatment. Flow cytometric analysis using Annexin V/PI staining revealed a significant increase in apoptotic cell populations (17.5%). Furthermore, gene expression analysis revealed marked upregulation of the pro-apoptotic genes, <i>Bax</i> (22.62-fold) and <i>Caspase-3</i> (22.06-fold), along with a downregulation of the anti-apoptotic gene, <i>Bcl-2</i> (0.235-fold), supporting apoptosis as a potential mechanism of cell death. These findings suggest that the CpsA–CpsC fusion protein may exert apoptotic effects in cervical cancer cells and provide a basis for further investigation of bacterial-derived components as candidate anticancer agents.</p>

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Streptococcus-derived CpsA–CpsC fusion protein induces apoptosis in cervical cancer cells through in silico and in vitro methods

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年5月30日

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  • Zeynab Marzhoseyni,
  • Mohammad Shayestehpour,
  • Zahra Shaghaghi,
  • Maryam Alvandi,
  • Davoud Esmaeili

摘要

Cervical cancer is a significant cause of cancer-related mortality in women, and the toxicity of conventional anticancer drugs has necessitated the development of novel therapeutic strategies. In the present study, we designed a fusion protein containing CpsA and CpsC proteins encoding capsular polysaccharides in Streptococcus agalactiae and evaluated its potential anticancer activity. Physicochemical properties, 3-dimensional (3D) structure, and molecular dynamics (MD) simulation were analyzed using in silico tools. Then, the recombinant gene was cloned, expressed in a bacterial host, and purified successfully. Finally, the anticancer effects of the CpsA-CpsC fusion protein on cervical cancer cell line (HeLa) were evaluated through MTT, Real-Time PCR, and flow cytometry techniques. Functional assays demonstrated that the CpsA–CpsC fusion protein exerted time- and dose-dependent cytotoxic effects on HeLa cells, with an IC50 value of 68.1 µg/mL at 48 after 48 h of treatment. Flow cytometric analysis using Annexin V/PI staining revealed a significant increase in apoptotic cell populations (17.5%). Furthermore, gene expression analysis revealed marked upregulation of the pro-apoptotic genes, Bax (22.62-fold) and Caspase-3 (22.06-fold), along with a downregulation of the anti-apoptotic gene, Bcl-2 (0.235-fold), supporting apoptosis as a potential mechanism of cell death. These findings suggest that the CpsA–CpsC fusion protein may exert apoptotic effects in cervical cancer cells and provide a basis for further investigation of bacterial-derived components as candidate anticancer agents.