Metabolic Syndrome Research Center, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Medical Genetics and Molecular Medicine, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran; Cancer Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
Metabolic Syndrome Research Center, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Medical Biochemistry, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Tissue Cell. 2022 Aug;77:101853. doi: 10.1016/j.tice.2022.101853. Epub 2022 Jun 15.
Colorectal-cancer (CRC) is amongst the most lethal-cancers, mainly due to its metastatic spread and drug chemoresistance. Hence there is a need for new approaches to either increase the efficacy of current therapy or introduce new therapies that have greater efficacy. There is increasing evidence that dysregulation of WNT-signaling-pathway plays an essential role in the development and prognosis of CRC. Here we have investigated the therapeutic potential of targeting the WNT/b-catenin pathway using a novel Wnt/b-catenin inhibitor, PNU-74654, in combination with 5-FU in CRC.
The anti-proliferative-effect of PNU-74654 was evaluated in two-/three-dimensional cell models. The activity of agents on cell growth, migration, invasion, cell cycle and apoptosis was evaluated by MTT, wound healing assay, invasion, FACS, and annexin V staining, respectively. The oxidant/antioxidant levels were also assessed by determining the level of MDA, SOD, as well as using the DCFH-DA assay. We used a xenograft model of CRC to investigate PNU-74654 activity alone and in combination with 5-FU follow by histological staining and biochemical and gene expression analyses by RT-PCR and western blot.
PNU-74654 inhibited cell-growth and synergistically affected the anti-tumor properties of 5-FU via modulation of Cyclin D1 and survivin. This agent inhibited the migration/invasion of colorectal cancer cells via perturbation of E-cadherin. Furthermore, PNU-74654 inhibited the tumor growth, which was more pronounced using the PNU-74654 plus 5-FU combination via induction of reactive oxygen species, down-regulation of SOD and modulation of MCP-1, P53, TNF-α.
Our finding demonstrated that PNU-74654 can target Wnt-pathway, interfere with cell-proliferation, induced-cell death, reduced-migration and interact with 5-FU, supporting further investigations on this therapeutic-approach for colorectal cancer.
结直肠癌（CRC）是最致命的癌症之一，主要是由于其转移扩散和药物化疗耐药性。因此，需要新的方法来提高现有治疗的疗效或引入更有效的新疗法。越来越多的证据表明，WNT 信号通路的失调在 CRC 的发展和预后中起着至关重要的作用。在这里，我们研究了使用新型 Wnt/b-连环蛋白抑制剂 PNU-74654 靶向 WNT/b-连环蛋白途径与 5-FU 联合治疗 CRC 的治疗潜力。
在二维/三维细胞模型中评估 PNU-74654 的抗增殖作用。通过 MTT、划痕愈合试验、侵袭试验、FACS 和 Annexin V 染色分别评估药物对细胞生长、迁移、侵袭、细胞周期和细胞凋亡的作用。还通过测定 MDA、SOD 的水平以及使用 DCFH-DA 测定法来评估氧化剂/抗氧化剂水平。我们使用 CRC 的异种移植模型来研究 PNU-74654 单独使用和与 5-FU 联合使用的活性，然后进行组织学染色以及通过 RT-PCR 和 Western blot 进行生化和基因表达分析。
PNU-74654 抑制细胞生长，并通过调节细胞周期蛋白 D1 和生存素协同影响 5-FU 的抗肿瘤特性。该药物通过干扰 E-钙粘蛋白来抑制结肠癌细胞的迁移/侵袭。此外，PNU-74654 抑制肿瘤生长，与 5-FU 联合使用时更为明显，这是通过诱导活性氧、下调 SOD 以及调节 MCP-1、P53、TNF-α 来实现的。
我们的研究结果表明，PNU-74654 可以靶向 Wnt 通路，干扰细胞增殖，诱导细胞死亡，减少迁移，并与 5-FU 相互作用，支持对这种治疗结直肠癌的方法进行进一步研究。
