Department of Advanced Medical Sciences & Technologies, School of Medicine, Jahrom University of Medical Sciences, Jahrom, Iran; Department of Nanobiotechnology, Faculty of Biological Sciences,Tarbiat Modares University, Tehran, Iran.
Department of Molecular Biology, Tofigh Daru Co. (TODACO), Tehran, Iran.
Biomed Pharmacother. 2023 Oct;166:115299. doi: 10.1016/j.biopha.2023.115299. Epub 2023 Aug 11.
Genomic DNA sequences provide unique target sites, with high druggability value, for treatment of genetically-linked diseases like cancer. B-cell lymphoma protein-2 (BCL-2) prevents Bcl-2-associated X protein (BAX) and Bcl-2 antagonist killer 1 (BAK) oligomerization, which would otherwise lead to the release of several apoptogenic molecules from the mitochondrion. It is also known that BCL-2 binds to and inactivates BAX and other pro-apoptotic proteins, thereby inhibiting apoptosis. BCL-2 protein family, through its role in regulation of apoptotic pathways, is possibly related to chemo-resistance in almost half of all cancer types including breast cancer. Here for the first time, we have developed a nanohybrid using a peptide-based carrier and a Deoxyribonucleic acid inhibitor (DNAi) against BCL-2 oncogene to induce apoptosis in breast cancer cells. The genetically designed nanocarrier was functionalized with an internalizing RGD (iRGD) targeting motif and successfully produced by recombinant DNA technology. Gel retardation assay demonstrated that the peptide-based carrier binds single-stranded DNAi upon simple mixing. Dynamic light scattering (DLS) and transmission electron microscopy (TEM) analyses further revealed the formation of nanohybrid particles with a size of 30 nm and a slightly positive charge. This hemocompatible nanohybrid efficiently delivered its contents into cancer cells using iRGD targeting moiety. Gene expression analysis demonstrated that the nanohybrids, which contained DNAi against BCL-2 proficiently suppressed the expression of this oncogene in a sequence specific manner. In addition, the nanohybrid, triggered release of cytochrome c (cyt c) and caspase3/7 activation with high efficiency. Although the DNAi and free nanocarrier were separately unable to affect the cell viability, the nanohybrid of 20 nM of DNAi showed outstanding antineoplastic potential, which was adjusted by the ratio of the MiRGD nanocarrier to DNAi. It should be noted that, the designed nanohybrid showed a suitable specificity profile and did not affect the viability of normal cells. The results suggest that this nanohybrid may be useful for robust breast cancer treatment through targeting the BCL-2 oncogene without any side effects.
基因组 DNA 序列为治疗癌症等遗传性疾病提供了具有高成药性价值的独特靶标。B 细胞淋巴瘤蛋白-2 (BCL-2) 可阻止 Bcl-2 相关 X 蛋白 (BAX) 和 Bcl-2 拮抗剂杀伤 1 (BAK) 寡聚化，否则这将导致几种凋亡原分子从线粒体释放。此外，BCL-2 还与 BAX 和其他促凋亡蛋白结合并使其失活，从而抑制细胞凋亡。BCL-2 蛋白家族通过其在调节凋亡途径中的作用，可能与包括乳腺癌在内的近一半癌症类型的化疗耐药性有关。在这里，我们首次开发了一种使用基于肽的载体和针对 BCL-2 癌基因的脱氧核糖核酸抑制剂 (DNAi) 的纳米杂化物，以诱导乳腺癌细胞凋亡。这种通过基因设计的纳米载体被内化 RGD (iRGD) 靶向基序功能化，并通过重组 DNA 技术成功生产。凝胶阻滞实验表明，基于肽的载体在简单混合时结合单链 DNAi。动态光散射 (DLS) 和透射电子显微镜 (TEM) 分析进一步揭示了纳米杂化物颗粒的形成，其粒径为 30nm，带轻微正电荷。这种血液相容性纳米杂化物通过 iRGD 靶向部分有效地将其内容物递送到癌细胞中。基因表达分析表明，含有针对 BCL-2 的 DNAi 的纳米杂化物能够以序列特异性方式有效地抑制该癌基因的表达。此外，纳米杂化物能够高效触发细胞色素 c (cyt c) 的释放和 caspase3/7 的激活。虽然 DNAi 和游离纳米载体单独使用时都不会影响细胞活力，但 DNAi 为 20nM 的纳米杂化物显示出出色的抗肿瘤潜力，这可以通过 MiRGD 纳米载体与 DNAi 的比例来调节。值得注意的是，设计的纳米杂化物显示出适当的特异性特征，并且不会影响正常细胞的活力。结果表明，这种纳米杂化物通过靶向 BCL-2 癌基因而不会产生任何副作用，可能对乳腺癌的治疗有用。
