Journal: Molecular Cancer

Article Title: CREB3L1 promotes tumor growth and metastasis of anaplastic thyroid carcinoma by remodeling the tumor microenvironment

doi: 10.1186/s12943-022-01658-x

Figure Lengend Snippet: CREB3L1 is involved in remodeling the stromal microenvironment of ATC. A Single - cell RNA-sequencing was used to examine the interaction between ATC cell subsets and cells in the tumor microenvironment. B The sphere formation assay was used to detect the growth of 8505C cells mixed with or without CAFs after CREB3L1 knockdown. *** P < 0.001 CAF + NC versus CAF + CREB3L1 - KD. C Schematic diagram of the experimental procedure. Equal numbers of DiI - labeled 8505C cells (red) and DiO - labeled CAFs (green) were mixed and implanted into the perivitelline space of each zebrafish. D The zebrafish xenograft model was employed to evaluate the CAF - mediated metastasis of 8505C cells after CREB3L1 knockdown. E Flow cytometry analysis of α - SMA - positive fibroblasts, after co - culture with 8505C cells. * P < 0.05, ** P < 0.01 versus the respective CAF + medium, CAF + NC, or CAF + CREB3L1 - KD. F Flow cytometry analysis of α - SMA positive fibroblasts in the sphere derived from the mixture of 8505C and CAFs. G Flow cytometry analysis of α - SMA - positive fibroblasts in the lung tissues of mice with ATC pulmonary metastasis. * P < 0.05 NC versus CREB3L1 - KD. H Counterstaining of Ki67 and α - SMA with antibodies that specifically reacted with the human (Ki67) and mouse (α - SMA) antigen, respectively. Data are presented as the mean ± SD

Article Snippet: The immortalized thyroid cancer - associated fibroblast (CAF) cell line (China Center for Type Culture Collection, C2022119) was isolated from a human PTC tumor and cultured in a DMEM/F12 (1:1) (Gibco) medium, supplemented with 10% FBS.

Techniques: RNA Sequencing Assay, Tube Formation Assay, Labeling, Flow Cytometry, Co-Culture Assay, Derivative Assay