Leveraging the htsFLT01/MiRGD Complex to Enhance Apoptosis and Suppress Angiogenesis in MCF7 Breast Cancer Cells

TL;DR

This study explores using a gene therapy complex to boost cell death and reduce blood vessel growth in breast cancer cells.

Contribution

The study introduces a novel gene therapy complex combining htsFLT01 and MiRGD for enhanced anti-cancer effects.

Findings

The htsFLT01/MiRGD complex increased expression of FADD, CASP8, and p53 genes in MCF7 cells.

The complex showed optimal performance at an N/P ratio of 14.

The therapy demonstrated a synergistic anti-angiogenic and pro-apoptotic effect.

Abstract

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.

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.

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.

These findings highlight a synergistic relationship between anti-angiogenic and pro-apoptotic mechanisms, offering promising avenues for future breast cancer therapies.

Figures