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Author Spotlight: Exploring Salidroside's Molecular Mechanisms in Breast Cancer Treatment
Published on: June 9, 2023
Dermaseptin B2 bioactive gene's potential for anticancer and anti-proliferative effect is linked to the regulation of
Fatemeh Khak-Rah Nasirabadi1, Abbas Doosti2
1Department of Biology, Faculty of Basic Sciences, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran.
Abstract:
Dermaseptin B2 (DrsB2) is an antimicrobial peptide with anticancer and angiostatic properties. We aimed to assess the in vitro inhibitory effect of pDNA/DrsB2 on the growth of breast cancer cells and its impact on the expression of genes involved in the BAX/BBC3/AKT pathway. The nucleic acid sequence of DrsB2 was artificially synthesized and inserted into the pcDNA3.1( +) Mammalian Expression Plasmid. PCR testing and enzyme digesting procedures evaluated the accuracy of cloning. The vectors were introduced into cells using LipofectamineTM2000 transfection reagent. The breast cancer cells were assessed by flow cytometry, MTT assessment, soft agar colony method, and wound healing investigation. The gene's transcription was evaluated using real-time PCR with a significance level of P < 0.05. The recombinant plasmid harboring the pDNA/DrsB2 vector was effectively produced, and the gene sequence showed absolute homogeneity (100% similarity) with the DrsB2 gene. The transfection effectiveness of MCF-7 and MCF-10A cells was 79% and 68%, respectively. The findings are measured using the growth inhibition 50% (GI50) metric, which indicates the concentration of pDNA/DrsB2 that stops 50% of cell growth. The proportions of early apoptosis, late apoptosis, necrosis, and viable MCF-7 cells in the pDNA/DrsB2 group were 40.50%, 2.31%, 1.69%, and 55.50%, respectively. The results showed a 100% increase in gene expression in programmed cell death following treatment with pDNA/DrsB2 (**P < 0.01). To summarize, the results described in this work offer new possibilities for treating cancer by targeting malignancies via pDNA/DrsB2 and activating the BAX/BBC3/AKT signaling pathways.
Insights
Dermaseptin B2 (DrsB2) effectively inhibits breast cancer cell growth in vitro. This antimicrobial peptide activates programmed cell death pathways, offering a potential new cancer treatment strategy.
Area of Science:
- Molecular Biology
- Biochemistry
- Oncology
Background:
- Dermaseptin B2 (DrsB2) is an antimicrobial peptide with demonstrated anticancer and angiostatic properties.
- The BAX/BBC3/AKT pathway is crucial in regulating programmed cell death and cancer progression.
Purpose of the Study:
- To investigate the in vitro inhibitory effect of a plasmid encoding DrsB2 (pDNA/DrsB2) on breast cancer cell growth.
- To analyze the impact of pDNA/DrsB2 on the expression of genes within the BAX/BBC3/AKT pathway.
Main Methods:
- Artificial synthesis and cloning of the DrsB2 gene into a mammalian expression plasmid (pcDNA3.1(+)).
- Transfection of breast cancer cells (MCF-7) and normal cells (MCF-10A) using Lipofectamine™2000.
- Assessment of cell viability and apoptosis via flow cytometry, MTT assay, soft agar, and wound healing assays.
- Quantification of gene expression changes using real-time PCR.
Main Results:
- Successfully constructed and verified the pDNA/DrsB2 recombinant plasmid with 100% gene sequence homogeneity.
- Achieved effective transfection rates in MCF-7 (79%) and MCF-10A (68%) cells.
- pDNA/DrsB2 treatment significantly induced apoptosis (40.50% early, 2.31% late) and increased programmed cell death gene expression by 100% (P < 0.01).
Conclusions:
- pDNA/DrsB2 demonstrates significant in vitro anticancer activity against breast cancer cells.
- The observed anticancer effects are associated with the activation of the BAX/BBC3/AKT signaling pathway, leading to programmed cell death.
- pDNA/DrsB2 represents a promising therapeutic candidate for targeting malignancies by modulating key cell death pathways.
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