Premium
Molecular Subtyping and In Vivo Phenotyping Studies Identify Human Cell Lines Suitable for Bladder Cancer Research
Author(s) -
Warrick Joshua,
Shuman Lauren,
Zheng Zongyu,
Yamashita Hironobu,
Amponsa Vasty Osei,
DeGraff David
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.445.3
Subject(s) - biology , phenotype , gene expression , cancer research , gene , gene expression profiling , subtyping , gata3 , pathology , genetics , transcription factor , medicine , computer science , programming language
Recent high profile studies have reported the subdivision of urothelial carcinoma (UC) of the bladder into molecular subtypes based upon gene expression and genetic alterations. The precise classification and naming differ by study, though all studies have identified at least two large molecular subtypes of UC, referred to as “luminal” and “basal”. Bladder UCs of the luminal subtype are enriched for mutations in fibroblast growth factor receptor 3 (FGFR3), and often exhibit with papillary morphology and increased expression of genes associated with urothelial differentiation. Conversely, the basal molecular subtype of human UC is enriched for mutations in TP53 and RB1, and often exhibits squamous morphology and expression of genes associated with a squamous phenotype. In order to determine if there exists an association between molecular subtype and phenotypic behavior, we sought to investigate best classification of these cells lines using publically available gene expression and genetic data, as well as morphologic and phenotypic analysis of tumors following tissue recombination xenografting experiments. Following extraction of expression data for 22 UC cell lines from the Cancer Cell Line Encyclopedia (CCLE), consensus cluster analysis revealed three main clusters: basal, luminal, and a third cluster with low expression of luminal and basal genes ( Figure). Expression profiles of the examined UC cell lines were diverse. RT4 was strongly luminal, with high expression of markers of urothelial differentiation (e.g. FOXA1, GATA3) and low expression of basal markers. SCaBER was strongly basal, with high expression of basal markers (e.g. CK14, EGFR) and low expression of luminal markers. UMUC1 had high expression of both luminal and basal markers, though it clustered with luminal cell lines in our analysis. Low expression of basal and luminal markers was seen in SWI1710, UMUC3, HS172T, TCCSUP, T24, JMSU1, and J82. A subgroup of cell lines (RT4, SCaBER, UMUC1, T24, HT1376, and 5637) were recombined with embryonic rat bladder mesenchyme (eBLM) and inserted into the kidney capsule of immunocompromised mice to identify potential association between molecular subtype, tumorigenicity, and histology. Interestingly, RT4, UMUC1, SCaBER, and HT1376 were highly tumorigenic, while T24 and 5637 cells failed to form tumors following surgical implantation. Much as is the case in human disease, molecular profiling correlated with the observed histologic phenotype of surgically implanted cell lines. For example, RT4 tumors cells exhibited papillary morphology, keeping with the luminal subtype, while SCaBER cells demonstrated keratin formation, keeping with the squamous phenotype of basal tumors. UMUC1 showed aggressive invasion, but no had papillary morphology or squamous differentiation. In summary, our findings identify human UC cell lines which represent the spectrum of human UC in regard to molecular subtype. We recommend RT4 as a model for luminal type bladder cancer, and SCaBER as a model for basal type bladder cancer with squamous differentiation.Expression subtypes of bladder cancer cell lines. Red=higher expression, green=lower expression.