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Designing a novel fusion protein from Streptococcus agalactiae with apoptosis induction effects on cervical cancer
Zeynab Marzhoseyni1, Mohammad Shayestehpour2, Morteza Salimian3
1Department of Microbiology and Immunology, Faculty of Medicine, Kashan University of Medical Sciences, Kashan, Iran.
Abstract:
Cervical cancer remains life-threatening cancer in women around the world. Due to the limitations of conventional treatment approaches, there is an urgent need to develop novel and more efficient strategies against cervical cancer. Therefore, the researchers attend to the alternative anti-cancer compounds like bacterial products. Rib and α are known as surface proteins of Streptococcus agalactiae with immunologic effects. In the present study, we designed a new anti-cancer fusion protein (Rib-α) originating from S. agalactiae with in silico methods, and then, the recombinant gene was cloned in the pET-22 (+) expression vector. The recombinant protein was expressed in E. coli BL21. To purify the expressed protein, we applied the Ni-NTA column. The molecular mechanism by which Rib-α is cytotoxic to cancer cells has been discussed based on MTT, flow cytometry, and real-time PCR methods. The engineered fusion protein suppressed the proliferation of the cancer cells at 180 μg/ml. Cytotoxic assessment and morphological changes, augmentation of apoptotic-related genes, upregulation of caspase-3 mRNA, and flow cytometric analysis confirmed that apoptosis might be the principal mechanism of cell death. According to our findings, Rib-α fusion protein motivated the intrinsic apoptosis pathway. Therefore, it can be an exciting candidate to discover a new class of antineoplastic agents.
Insights
A novel bacterial fusion protein, Rib-α, effectively suppresses cervical cancer cell proliferation. This engineered protein induces apoptosis, presenting a promising new avenue for developing anti-cancer therapies.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Cervical cancer poses a significant global health threat to women worldwide.
- Conventional treatments for cervical cancer have limitations, necessitating novel therapeutic strategies.
- Bacterial products are being explored as alternative anti-cancer agents due to their immunologic effects.
Purpose of the Study:
- To design and characterize a novel anti-cancer fusion protein, Rib-α, derived from Streptococcus agalactiae surface proteins.
- To investigate the molecular mechanisms underlying the cytotoxic effects of Rib-α on cancer cells.
- To evaluate the potential of Rib-α as a new therapeutic candidate for cervical cancer treatment.
Main Methods:
- In silico design and recombinant gene cloning of the Rib-α fusion protein.
- Expression and purification of the recombinant Rib-α protein in E. coli.
- Assessment of cytotoxicity using MTT assays, flow cytometry, and real-time PCR to analyze gene expression and apoptosis.
Main Results:
- The engineered Rib-α fusion protein demonstrated significant suppression of cancer cell proliferation at a concentration of 180 μg/ml.
- Cytotoxic assessments, morphological changes, and flow cytometry confirmed apoptosis as the primary mechanism of cell death.
- Real-time PCR analysis revealed the upregulation of apoptosis-related genes, including caspase-3 mRNA.
Conclusions:
- The Rib-α fusion protein effectively induces apoptosis in cancer cells, primarily through the intrinsic apoptosis pathway.
- Rib-α exhibits potential as a novel anti-cancer agent for cervical cancer.
- This study highlights the promise of bacterial-derived fusion proteins in the development of new antineoplastic therapies.
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