胰腺癌(PC)的预后特征取决于肿瘤微环境的影响因素。瘦素是脂肪细胞作为脂肪因子释放到肿瘤微环境中的关键致癌因子。它通过激活癌症干细胞(CSC)分子调节剂Notch来促进肿瘤的发展,刺猬,和Wnt/β-连环蛋白信号传导。这些途径的下游靶标之一是CDK4/6和细胞周期蛋白D,其由在PC中高度突变的P16INK4A控制。因此,这项研究的目的是确定CDK4/6抑制剂的作用,palbociclib,在瘦素诱导的PC细胞上,并靶向Notch,刺猬,和Wnt/β-连环蛋白信号通路通过miR-150、miR-506和miR-208调节。瘦素治疗增加了Panc-1,MiaPaCa-2和Capan-2细胞的增殖能力,并降低了palbociclib的作用。此外,从瘦素处理的(瘦素)和瘦素未处理的(瘦素-)Panc-1和MiaPaCa-2细胞产生肿瘤球,并用miR-506,miR-150(肿瘤抑制miRNA)转染,或抗miR-208(oncomiR),其次是palbociclib治疗。miR-506或miR-150的强制表达通过抑制集落和肿瘤球状体的形成,显著增加了瘦素+细胞对palbociclib治疗的易感性,和CD44在Panc-1和MiaPaCa-2细胞中的表达。此外,增加的miR-150表达更有效地抑制N-cadherin,β-连环蛋白,p-GSK3β,缺口,和Wnt5a/b在Leptin-/+Panc-1和MiaPaCa-2细胞中的表达。因此,palbociclib抑制了瘦素治疗诱导的CSC谱,抑制肿瘤球形式和瘦素靶向信号通路,从而禁用Panc-1和MiaPaCa-2细胞的抗性机制。miR-506或miR-150的表达增加和miR-208的抑制增强了Panc-1和MiaPaCa-2Leptin-/+细胞对palbociclib治疗的敏感性。因此,这项研究证明,将CSC分子调节剂抑制剂与palbociclib联合使用可提高抑制PC细胞增殖的成功率。
The prognostic characteristics of pancreatic cancer (PC) are determined by the contributing factors from the tumor microenvironment. Leptin is a critical oncogenic factor released by adipocytes as an adipokine into the tumor microenvironment, where it promotes tumor development by activating cancer stem cell (CSC) molecular regulators Notch, Hedgehog, and Wnt/β-catenin signaling. One of the downstream targets of these pathways is CDK4/6 and cyclin D which is controlled by P16 INK4A that is highly mutated in PC. Therefore, the purpose of this study was to determine the effect of a CDK4/6 inhibitor, palbociclib, on Leptin-induced PC cells and to target the Notch, Hedgehog, and Wnt/β-catenin signaling pathways via miR-150, miR-506, and miR-208 modulation. Leptin treatment increased the ability of Panc-1, MiaPaCa-2, and Capan-2 cells to proliferate and decreased the effect of palbociclib. Additionally, tumorspheres were generated from Leptin-treated (Leptin+) and Leptin-untreated (Leptin-) Panc-1 and MiaPaCa-2 cells and transfected with miR-506, miR-150 (tumorsuppressor miRNAs), or anti-miR-208 (oncomiR), followed by palbociclib treatment. Forced expression of miR-506 or miR-150 significantly increased the susceptibility of Leptin+ cells to palbociclib treatment by inhibiting colony and tumor spheroid formation, and CD44 expression in Panc-1 and MiaPaCa-2 cells. Additionally, the increased miR-150 expression is more effective at inhibiting N-cadherin, β-catenin, p-GSK3β, Notch, and Wnt5a/b expression in Leptin-/+ Panc-1 and MiaPaCa-2 cells. As a result, palbociclib suppressed the CSC profile induced by leptin treatment, inhibiting both tumorsphere forms and leptin-targeted signaling pathways, thereby disabling the Panc-1 and MiaPaCa-2 cells\' resistance mechanism. Increased expression of miR-506 or miR-150 and inhibition of miR-208 enhanced sensitivity of Panc-1 and MiaPaCa-2 Leptin-/+ cells to palbociclib treatment. As a result, this study proved that combining inhibitors of CSC molecular regulators with palbociclib improves the success rate of inhibition of PC cell proliferation.