Cancer-Associated Fibroblasts induce aggressive stem cell phenotype and chemoresistance in cervical tumoroid-fibroblast co-culture system via upregulating CK17 expression

Cervical cancer is a serious health issue affecting women. Cancer development is the result of co-evolution between tumor cells and tumor stroma microenvironment, and Cancer Associated Fibroblasts (CAFs) is a significant part of tumor stroma and plays a big role in helping cancer cells grow, spread, and become aggressive. To investigate the biological function of CAFs in the progression of cervical cancer, we developed a novel CAFs-integrated cervical tumoroid model that can mimic the tumor microenvironment. Our cultured cervical organoids successfully reproduced key features of cervical tissue epithelium, including cellular composition, histological properties and biomarker distribution. Within the co-culture model, CAFs were found to stimulate cancer cells to acquire an aggressive, stem cell-like phenotype. Additionally, CAFs induced an elevated secretion of collagen I, which altered the extracellular matrix to facilitate tumor metastasis. Of particular interest, we discovered that CAFs modulate the CK17-related signaling pathway, causing upregulation of a suite of genes involved in Epithelial-Mesenchymal Transition (EMT) and Cancer Stem Cell (CSC), thus significantly boosting the invasive capacity of tumor cells. Both CK17-overexpressing tumoroids and those co-cultured with CAFs exhibited similar invasive phenotypes in type I collagen matrices, outperforming control tumoroids in terms of invasiveness and proliferation.Our findings indicate that CAFs likely enhance chemoresistance in tumoroid and contribute to malignant progression partly through CK17 upregulation. This implicates CK17 could be a promising clinical target for cervical cancer prevention and treatment.