Department of Genetic Engineering, School of Bio-Engineering, SRM Institute of Science and Technology, Kattankulathur, Kanchipuram 603203, Tamil Nadu, India.
Department of Endocrinology, Dr. ALM PG Institute of Basic Medical Sciences, University of Madras, Taramani Campus, Chennai 600113, Tamil Nadu, India.
Colloids Surf B Biointerfaces. 2022 Sep;217:112612. doi: 10.1016/j.colsurfb.2022.112612. Epub 2022 Jun 7.
The cancer stem cell (CSC) hypothesis is an evolving oncogenesis concept. CSCs have a distinct ability to self-renew themselves and also give rise to a phenotypically diverse population of cells. Targeting CSCs represents a promising strategy for cancer treatment. Plant-derived compounds are potent in restricting the expansion of CSCs. DCLK1 has been already reported as a colon CSC specific marker. Nanoparticles can effectively inhibit multiple types of CSCs by targeting specific markers. We have synthesized DCLK1 functionalized folic acid conjugated hesperetin encapsulated chitosan nanoparticles (CFH-DCLK1), specifically to target CSCs. In this regard, we have performed proliferation assay, colony formation assay, cell migration assay, apoptosis assay, flow cytometry analysis, real-time RT- PCR and western blot analyses to determine the effect of CFH-DCLK1 and CFH nanoparticles in HCT116-colon cancer cells. In our study, we have determined the median inhibitory concentration (IC50) of CFH (47.8 µM) and CFH-DCLK1 (4.8 µM) nanoparticles in colon cancer cells. CFH-DCLK1 nanoparticles induced apoptosis and inhibited the migration and invasion of colon cancer cells. Real time PCR and western blot results have demonstrated that the treatment with CFH-DCLK1 nanoparticles significantly reduced the expression of CSC markers such as DCLK1, STAT1 and NOTCH1 compared to the CFH alone in HCT116 colon cancer cells. Finally, in the 3D spheroid model, CFH-DCLK1 nanoparticles significantly inhibited the colonosphere growth. Overall, our results highlight the effectiveness of CFH-DCLK1 nanoparticles in targeting the colon cancer cells and CSCs. This study would lead to the development of therapies targeting both cancer cells and CSCs simultaneously using nanoformulated drugs, which could bring changes in the current cancer treatment strategies.
癌症干细胞（CSC）假说，是一个不断发展的肿瘤发生概念。CSC 具有独特的自我更新能力，并且还产生表型多样化的细胞群体。靶向 CSC 代表了一种有前途的癌症治疗策略。植物衍生化合物在限制 CSC 的扩增方面非常有效。DCLK1 已被报道为结肠 CSC 特异性标志物。纳米粒子可以通过靶向特定标志物来有效抑制多种类型的 CSC。我们已经合成了 DCLK1 功能化叶酸偶联橙皮苷封装壳聚糖纳米粒子（CFH-DCLK1），专门用于靶向 CSC。在这方面，我们进行了增殖实验、集落形成实验、细胞迁移实验、细胞凋亡实验、流式细胞术分析、实时 RT-PCR 和 Western blot 分析，以确定 CFH-DCLK1 和 CFH 纳米粒子在 HCT116 结肠癌细胞中的作用。在我们的研究中，我们确定了 CFH（47.8µM）和 CFH-DCLK1（4.8µM）纳米粒子在结肠癌细胞中的中值抑制浓度（IC50）。CFH-DCLK1 纳米粒子诱导结肠癌细胞凋亡，并抑制其迁移和侵袭。实时 PCR 和 Western blot 结果表明，与单独使用 CFH 相比，CFH-DCLK1 纳米粒子处理显著降低了 HCT116 结肠癌细胞中 CSC 标志物如 DCLK1、STAT1 和 NOTCH1 的表达。最后，在 3D 球体模型中，CFH-DCLK1 纳米粒子显著抑制了结肠球体的生长。总之，我们的研究结果强调了 CFH-DCLK1 纳米粒子在靶向结肠癌细胞和 CSC 方面的有效性。这项研究将导致使用纳米制剂药物同时靶向癌细胞和 CSC 的治疗方法的发展，这可能会改变当前的癌症治疗策略。
