Department of Medical Biotechnology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran.
Cancer Gene Therapy Research Center, Zanjan University of Medical Sciences, Zanjan, Iran.
Mol Biol Rep. 2020 Mar;47(3):1859-1869. doi: 10.1007/s11033-020-05280-2. Epub 2020 Feb 3.
Cancer stem cells are commonly tolerant toward chemotherapy and radiotherapy. Oct4 and Sox2 transcription factors are shown to be overexpressed in various cancers. At the current research, inhibition of Oct4 and Sox2 transcription factors was performed through application of decoy oligodeoxynucleotides (ODNs) strategy via repressing stemness properties in HT29-ShE cells encompassing enriched cancer stem-like cells. Designed Oct4-Sox2 complex decoy ODNs were transfected into HT29-ShE cells with Lipofectamine reagent. At the next step, ODNs efficiency transfection and subcellular localization were determined via flow cytometry and fluorescence microscopy, respectively. Further investigations such as cell proliferation and apoptosis analysis, colonosphere formation, invasion and migration, and real-time PCR assays were also carried out. Obtained results shed light on the fact that the designed complex decoys were effectively transfected into HT29-ShE cells, and they were found to be localized in subcellular compartments. Oct4-Sox2 decoy ODNs led to decreased cell viability, arresting the cell cycle in G0/G1 phases, increasing apoptosis, inhibition of migration/invasion and colonosphere formation ability of HT29-ShE cells in comparison with control and scramble groups. Furthermore, Oct4-Sox2 complex decoy could modulate the MET process via alteration of mRNA expression of downstream genes. It could be concluded that application of Oct4-Sox2 transcription factor decoy strategy in cells with stemness potential could lead to inhibiting the cell growth and triggering differentiation. Therefore, this technique could be applied along with usual remedies (chemotherapy and radiotherapy) as high potential method for treating cancer.
肿瘤干细胞通常对化疗和放疗具有耐受性。Oct4 和 Sox2 转录因子在各种癌症中表达过度。在目前的研究中,通过应用诱饵寡脱氧核苷酸 (ODN) 策略抑制 Oct4 和 Sox2 转录因子,从而抑制包含丰富肿瘤干细胞样细胞的 HT29-ShE 细胞的干性特性。设计的 Oct4-Sox2 复合物诱饵 ODN 通过脂质体试剂转染 HT29-ShE 细胞。下一步,通过流式细胞术和荧光显微镜分别确定 ODN 的转染效率和亚细胞定位。还进行了细胞增殖和凋亡分析、类器官形成、侵袭和迁移以及实时 PCR 测定等进一步研究。结果表明,设计的复合物诱饵被有效转染到 HT29-ShE 细胞中,并发现它们定位于亚细胞区室中。与对照组和乱序组相比,Oct4-Sox2 诱饵 ODN 导致 HT29-ShE 细胞活力降低,细胞周期停滞在 G0/G1 期,凋亡增加,迁移/侵袭抑制和类器官形成能力降低。此外,Oct4-Sox2 复合物诱饵可以通过改变下游基因的 mRNA 表达来调节 MET 过程。可以得出结论,在具有干细胞特性的细胞中应用 Oct4-Sox2 转录因子诱饵策略可能导致抑制细胞生长和触发分化。因此,该技术可以与常规治疗方法(化疗和放疗)一起作为治疗癌症的高潜力方法。