Valouzi Atefeh, Shahbazi Majid, Erfani-Moghadam Vahid, Ramezani Mahboobeh, Shamsabadi Fatemeh T
Department of Medical Biotechnology, Golestan University of Medical Sciences, Gorgan, Iran.
Medical Cellular & Molecular Research Center, Golestan University of Medical Sciences, Gorgan, Iran.
Mol Biotechnol. 2025 Jan 17. doi: 10.1007/s12033-024-01359-4.
Oncolytic viral-based therapy and specific gene expression by promoters are modern targeted oncotherapy approaches that have gained significant attention in recent years. In this study, both strategies were combined by designing cancer-specific activation of vesicular stomatitis virus matrix expression under the survivin promoter. The matrix sequence was cloned downstream of the survivin promoter (pM). After transfecting MCF-7 cells with pM, cell proliferation and apoptosis induction were assessed. Additionally, the transcript levels of matrix and apoptosis-related genes in response to pM was assessed. The proliferation of MCF-7 cells was significantly reduced by the constructed matrix-expressing plasmid at 48 and 72 h post-transfection (p < 0.05). Enhanced matrix expression resulted in the down-regulation of MMP-9, TP53, and NF-kB, while simultaneously up-regulating Bax transcripts. Evaluating the effect of pM vector on apoptosis induction revealed a significant increase in the MCF-7 cells compared to untreated cells (p < 0.05). The absence of significant matrix gene expression in HDF cells, relative to MCF-7 cells, further underscores the specific function of the Survivin promoter in cancer cells. These findings suggest that the matrix may have various biological functions in a diverse set of non-apoptotic pathways. Further research on the association of the matrix with other genes could provide insights into the biomedical significance and future perspectives of the matrix in cancer gene therapy.
基于溶瘤病毒的疗法和启动子介导的特定基因表达是近年来备受关注的现代靶向肿瘤治疗方法。在本研究中，通过设计在生存素启动子调控下特异性激活水疱性口炎病毒基质蛋白的表达，将这两种策略结合起来。将基质蛋白序列克隆到生存素启动子（pM）的下游。用pM转染MCF-7细胞后，评估细胞增殖和凋亡诱导情况。此外，还评估了pM作用下基质蛋白和凋亡相关基因的转录水平。构建的表达基质蛋白的质粒在转染后48小时和72小时显著降低了MCF-7细胞的增殖（p < 0.05）。基质蛋白表达增强导致MMP-9、TP53和NF-kB下调，同时上调Bax转录本。评估pM载体对凋亡诱导的作用发现，与未处理的细胞相比，MCF-7细胞的凋亡显著增加（p < 0.05）。与MCF-7细胞相比，人皮肤成纤维细胞（HDF）中基质蛋白基因无明显表达，这进一步强调了生存素启动子在癌细胞中的特异性功能。这些发现表明，基质蛋白可能在多种非凋亡途径中具有多种生物学功能。进一步研究基质蛋白与其他基因的关联，可为基质蛋白在癌症基因治疗中的生物医学意义和未来前景提供见解。
