Document Type : Original Article(s)

Authors

• Mohadeseh Khoshandam ¹
• Zahra-Soheila Soheili ¹
• Saman Hosseinkhani ²
• Shahram Samiee ³
• Hamid Latifi-Navid ¹
• Naser Kalhor ⁴
• Hossein Soltaninejad ⁵

¹ Department of Molecular Medicine, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran

² Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran

³ Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran

⁴ Department of Cell Biology and Regenerative Medicine, Academic Center for Education, Culture and Research, Qom Branch, Qom, Iran

⁵ Department of stem cells technology and Tissue Regeneration, Faculty of Interdisciplinary Science and Technologies, Tarbiat Modares University, Tehran, Iran

Abstract

Background: Gene therapy introduces therapeutic genes into cancer cells to inhibit tumor growth or induce apoptosis. The htsFLT01 gene, a novel anti-angiogenic construct, encodes the sFLT01 protein that functions as a Vascular Endothelial Growth Factor (VEGF) decoy receptor, impeding pathological angiogenesis. When combined with the MiRGD nanocarrier—a versatile peptide-based delivery system optimized for specificity, biocompatibility, and low toxicity—the htsFLT01/MiRGD complex offers a potent strategy against breast cancer.
Methods: The htsFLT01 gene was designed and constructed in previous studies. The MiRGD peptide was expressed and purified using Ni-NTA affinity chromatography in the E. coli C41 (DE3) expression strain. The potency of this peptide, along with the cell viability and toxicity of the nanoparticles, was previously evaluated in MCF7 cell culture. After transfection with the htsFLT01/MiRGD nanocomplex at a nitrogen-to-phosphorus (N/P) ratio of 14, cell lysates were collected, and expression analysis of the key genes, including Fas-Associated Death Domain Protein (FADD), Caspase-8 (CASP8), and Tumor Protein P53 (TP53), was conducted based on findings from prior research. Statistical analyses were conducted using IBM SPSS Statistics version 22 (IBM, USA) and REST 2009 software.  
Results: The htsFLT01 gene was previously designed and constructed, and the MiRGD nanocarrier was successfully produced and purified. This nanocarrier exhibited the best performance at an N/P ratio of 14. This study evaluated the effect of this complex on apoptosis induction in MCF7 cells via the extrinsic apoptotic pathway, revealing increased expression of FADD, CASP8, and p53 genes. 
Conclusion: These findings highlight a synergistic relationship between anti-angiogenic and pro-apoptotic mechanisms, offering promising avenues for future breast cancer therapies.

Highlights

Mohadeseh Khoshandam (Google Scholar)

Zahra-Soheila Soheili (Google Scholar)

Keywords

• CASP8 and FADD-like apoptosis regulating protein
• Tumor suppressor protein p53
• Breast neoplasms
• Apoptosis
• Angiogenesis inhibitors