Pizzuto Elena, Mancarella Serena, Gigante Isabella, Serino Grazia, Dituri Francesco, Piccinno Emanuele, Fabregat Isabel, Giannelli Gianluigi
Personalized Medicine Laboratory, National Institute of Gastroenterology "S. de Bellis" IRCCS Research Hospital, 70013 Castellana Grotte, Italy.
TGF-β and Cancer Group-Oncobell Program, Bellvitge Biomedical Research Institute (IDIBELL), 08860 L'Hospitalet de Llobregat, Spain.
Int J Mol Sci. 2025 May 22;26(11):4973. doi: 10.3390/ijms26114973.
Molecular mechanisms responsible for the poor prognosis in patients with intrahepatic cholangiocarcinoma (CCA) are still unknown, but stem cell marker Cluster Differentiation 90 (CD90) has been reported to be associated with a more aggressive cancer phenotype. In this scenario, the TGF-β1 signaling pathway likely has a role as master gene regulator. Aim of the study is to investigate the role of CD90 in iCCA aggressiveness. The molecular profile of HuCCT1/CD90+ and HuCCT1/CD90- cells was obtained through transcriptomic analysis (NGS). Bioinformatic data were confirmed in both cell lines by qRT-PCR and Western blot. Cells were treated with Gemcitabine in monotherapy or in combination with Galunisertib, a selective inhibitor of TGF-βRI, in 2D and 3D models. HuCCT1/CD90+ cells are more proliferative, less migratory, and resistant to Gemcitabine treatment. HuCCT1/CD90+ cells also express lower levels of TGF-β1 compared to /CD90- cell lines. Finally, HuCCT1/CD90+ cells are resistant to Gemcitabine, while the combination of Gemcitabine and Galunisertib displays a synergistic effect on HuCCT1/CD90+ cell proliferation. These results underline that CD90-induced Gemcitabine resistance can be overcome by adding a TGFβ1 inhibitor such as Galunisertib, thereby moving further toward a precision medicine approach in patients with iCCA.
肝内胆管癌（CCA）患者预后不良的分子机制尚不清楚，但据报道干细胞标志物分化簇90（CD90）与更具侵袭性的癌症表型相关。在这种情况下，转化生长因子-β1（TGF-β1）信号通路可能作为主要基因调节因子发挥作用。本研究的目的是探讨CD90在肝内胆管癌侵袭性中的作用。通过转录组分析（NGS）获得了HuCCT1/CD90+和HuCCT1/CD90-细胞的分子图谱。通过qRT-PCR和蛋白质免疫印迹在两种细胞系中证实了生物信息学数据。在二维和三维模型中，细胞分别接受吉西他滨单药治疗或与TGF-βRI选择性抑制剂加鲁尼西单抗联合治疗。HuCCT1/CD90+细胞增殖能力更强，迁移能力较弱，且对吉西他滨治疗有抗性。与/CD90-细胞系相比，HuCCT1/CD90+细胞中TGF-β1的表达水平也较低。最后，HuCCT1/CD90+细胞对吉西他滨有抗性，而吉西他滨和加鲁尼西单抗的联合使用对HuCCT1/CD90+细胞增殖具有协同作用。这些结果强调，添加TGFβ1抑制剂如加鲁尼西单抗可以克服CD90诱导的吉西他滨抗性，从而在肝内胆管癌患者中进一步朝着精准医疗的方向发展。
