Dermaseptin B2 生物活性基因的抗癌和抗增殖潜力与 BAX/BBC3/AKT 通路的调节有关。

IF 2.8 4区 医学 Q2 ONCOLOGY Medical Oncology Pub Date : 2024-05-20 DOI:10.1007/s12032-024-02384-8
Fatemeh Khak-Rah Nasirabadi, Abbas Doosti
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引用次数: 0

摘要

Dermaseptin B2(DrsB2)是一种具有抗癌和血管抑制特性的抗菌肽。我们旨在评估 pDNA/DrsB2 对乳腺癌细胞生长的体外抑制作用及其对 BAX/BBC3/AKT 通路相关基因表达的影响。人工合成 DrsB2 的核酸序列并将其插入 pcDNA3.1( +) 哺乳动物表达质粒中。PCR 检测和酶解程序评估了克隆的准确性。使用 LipofectamineTM2000 转染试剂将载体导入细胞。通过流式细胞术、MTT 评估、软琼脂集落法和伤口愈合调查对乳腺癌细胞进行评估。使用实时 PCR 对基因转录进行评估,显著性水平为 P
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Dermaseptin B2 bioactive gene's potential for anticancer and anti-proliferative effect is linked to the regulation of the BAX/BBC3/AKT pathway.

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.

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来源期刊
Medical Oncology
Medical Oncology 医学-肿瘤学
CiteScore
4.20
自引率
2.90%
发文量
259
审稿时长
1.4 months
期刊介绍: Medical Oncology (MO) communicates the results of clinical and experimental research in oncology and hematology, particularly experimental therapeutics within the fields of immunotherapy and chemotherapy. It also provides state-of-the-art reviews on clinical and experimental therapies. Topics covered include immunobiology, pathogenesis, and treatment of malignant tumors.
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