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Investigating the effects of pcDNA3 polybia-MP1 on the apoptosis induction in lung cancer cell line
1Department of Biology, ShK.C., Islamic Azad University, Shahrekord, Iran.
Abstract:
Polybia-MP1 is an antimicrobial peptide (AMP) with significant anticancer efficacy. The aim of this study is to investigate the changes in the expression levels of the long non-coding RNAs (lncRNAs) MALAT1, LUCAT1, and BANCR in lung cancer cells after stimulation with pDNA/Polybia-MP1 complexes. This research seeks to understand the molecular mechanisms underlying Polybia-MP1's anticancer effects, specifically focusing on the involvement of these lncRNAs in lung cancer pathology and treatment response. The nucleic acid sequence of polybia-MP1 was inserted into the pcDNA3.1(+) Mammalian Expression Plasmid. The correctness of cloning was evaluated using PCR and enzyme digesting methodologies. The vectors were introduced into cells by transfection using LipofectamineTM2000. The A549 cancer cells were assessed using flow cytometry and wound healing studies. The expression levels of lncRNAs and apoptotic genes were evaluated using real-time PCR, with a significance threshold of P < 0.05. The pDNA/polybia-MP1 vector was effectively produced, and the gene sequence exhibited absolute consistency (100% similarity) with the polybia-MP1 gene. The proportions of early apoptosis, late apoptosis, necrosis, and viable A549 cells in the pDNA/polybia-MP1 group were 26%, 17%, 3%, and 54%, respectively. The RT-PCR analysis revealed that the introduction of pDNA/polybia-MP1 into A549 cells led to a reduction in the expression of PCA3, MALAT1, and LUCAT1 lncRNAs, as compared to the control group treated with PBS. Moreover, it increased the transcription of BANCR long non-coding RNA. The results showed a significant increase in the activity of transcription factors involved in programmed cell death after treatment with pDNA/polybia-MP1 (**P < 0.01). The study's findings suggest that pDNA/polybia-MP1 has the potential to significantly alter gene transcription in cancer cells, particularly concerning lncRNAs engaged in cell apoptotic pathways. The pDNA/polybia-MP1 compound, with its potent anticancer properties, has the capacity to induce apoptosis in cells, thereby offering a promising avenue for cancer treatment.
Insights
Polybia-MP1 peptide complexes effectively induce apoptosis in lung cancer cells by altering long non-coding RNA expression, offering a novel therapeutic strategy. This research highlights the potential of Polybia-MP1 in cancer treatment.
Area of Science:
- Molecular Biology
- Oncology
- Biochemistry
Background:
- Antimicrobial peptides (AMPs) like Polybia-MP1 show promise as anticancer agents.
- Long non-coding RNAs (lncRNAs) play crucial roles in cancer development and progression.
- Understanding the molecular mechanisms of Polybia-MP1's anticancer effects is vital for therapeutic development.
Purpose of the Study:
- To investigate the impact of pDNA/Polybia-MP1 complexes on lncRNA expression (MALAT1, LUCAT1, BANCR) in lung cancer cells.
- To elucidate the role of these lncRNAs in the anticancer activity of Polybia-MP1.
- To explore the potential of Polybia-MP1 as a targeted lung cancer therapy.
Main Methods:
- Construction and verification of pDNA/Polybia-MP1 expression vector using PCR and enzyme digestion.
- Transfection of A549 lung cancer cells with pDNA/Polybia-MP1 using LipofectamineTM2000.
- Assessment of apoptosis and cell viability via flow cytometry and wound healing assays.
- Quantification of lncRNA and apoptotic gene expression using real-time PCR.
Main Results:
- Successful construction of a consistent pDNA/Polybia-MP1 vector.
- pDNA/Polybia-MP1 treatment significantly increased apoptosis (early: 26%, late: 17%) and reduced viable A549 cells (54%).
- RT-PCR revealed decreased expression of MALAT1 and LUCAT1, and increased expression of BANCR, alongside enhanced transcription factors for programmed cell death (P < 0.01).
Conclusions:
- pDNA/Polybia-MP1 complexes effectively induce apoptosis in A549 lung cancer cells.
- The anticancer effects are associated with significant modulation of specific lncRNA expression.
- Polybia-MP1 demonstrates potential as a therapeutic agent for lung cancer by targeting lncRNA pathways.
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