Enhanced metastatic potential in the MB49 urothelial carcinoma model

Abstract

Recent data suggest that patients with a basal/stem-like bladder cancer (BC) subtype tend to have metastatic disease, but this is unconfirmed. Here we report the identification of murine MB49 cell line sub-clones with stem-like characteristics in culture. Subcutaneous implantation of S2 and S4 MB49 sub-clones into immunocompetent mice resulted in lung metastases in 50% and 80% of mice respectively, whereas none of the mice implanted with the parental cells developed metastasis. Gene profiling of cells cultured from S2 and S4 primary and metastatic tumors revealed that a panel of genes with basal/stem-like/EMT properties is amplified during metastatic progression. Among them, ITGB1 , TWIST1 and KRT6B are consistently up-regulated in metastatic tumors of both MB49 sub-clones. To evaluate clinical relevance, we examined these genes in a human public dataset and found that ITGB1 and KRT6B expression in BC patient tumor samples are positively correlated with tumor grade. Likewise, the expression levels of these three genes are correlated with worse clinical outcomes. This MB49 BC metastatic pre-clinical model provides a unique opportunity to validate and recapitulate results discovered in patient studies and to pursue future mechanistic therapeutic interventions for BC metastasis.

<p>HIF-1α, TWIST-1 and ITGB-1, associated with Tumor Stiffness, as Novel Predictive Markers for the Pathological Response to Neoadjuvant Chemotherapy in Breast Cancer</p>

“…In addition, over-expression of both TWIST-1 and ITGB-1 is associated with increased tumorigenesis, growth, metastasis, and resistance to chemotherapy in several cancers. 42,43 Based on the analysis above, we propose a hypothesis to explain why high tumor stiffness is closely correlated with chemoresistance in BC. The IFP increases in tumor environments with high matrix stiffness, leading to heterogeneity and tortuosity of neovascularization.…”

Bladder Cancer Metastasis Induced by Chronic Everolimus Application Can Be Counteracted by Sulforaphane In Vitro

“…Likewise, high expression of the integrin β1 subtype positively correlated with tumor grade and poor clinical outcome in bladder cancer patients [ 29 , 30 ]. Prevention of αV, α6, and β1 up-regulation by SFN or the SFN-everolimus combination could, therefore, immobilize tumor cells to impede cancer progression.…”

Non-stem bladder cancer cell-derived extracellular vesicles promote cancer stem cell survival in response to chemotherapy

“…The parental MB49 cells were a gift from Dr. Timothy Ratliff at Purdue University College of Veterinary Medicine and were cultured in RPMI 1640 (Thermo Fisher, USA) containing 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (Gibco, USA). The MB49 F1 and S2 sub-clones were established via the isolation of a single clone from MB49 primary cells as described previously [ 18 ]. Cisplatin (CDDP) was purchased from Sigma (USA).…”

“…RNA was extracted from MB49 cells as reported previously [ 18 ]. Briefly, cells that had reached 80–90% confluence in 100-mm dishes were lysed with 1 ml Trizol (Invitrogen, USA).…”