Chromosomal and proteome analysis of a new T24-based cell line model for aggressive bladder cancer.
Cell line models aid in understanding cancer aggressiveness. The aim of this study was the establishment of a metastatic variant (T24M) of the T24 bladder cancer cell line and its initial characterization at chromosomal and proteomic levels. T24M were spontaneously developed in mice from T24 cells, following cycles of subcutaneous injections and culture in vitro. Transwell migration assays and injections in mice revealed increased migration and tumorigenic properties of T24M compared to the T24 cells. Cytogenetic analysis demonstrated that T24M retained several karyotypic characteristics of the parental cells and also acquired novel chromosomal aberrations related to aggressive bladder cancer. Proteomic analysis of the T24 and T24M cells by 2-DE and MS led to the generation of their 2-DE proteomic map and revealed differences in multiple proteins. These include proteases of the lysosomal and proteasome degradation pathways, mitochondrial and cytoskeletal proteins. The 2-DE findings were confirmed by immunoblotting of cell lysates and immunohistochemistry of bladder cancer tissue sections for cathepsin D and activity assays for proteasome. Collectively, our results suggest that the T24M cells reflect many known chromosomal and proteomic aberrations encountered in aggressive bladder cancers but also provide access to novel findings with potentially clinical applications.
Makridakis
Manousos
M
Biotechnology Division, Biomedical Research Foundation, Academy of Athens, Athens, Greece.
Gagos
Sarantis
S
Petrolekas
Andreas
A
Roubelakis
Maria G
MG
Bitsika
Vasiliki
V
Stravodimos
Konstantinos
K
Pavlakis
Kitty
K
Anagnou
Nicholas P
NP
Coleman
Jonathan
J
Vlahou
Antonia
A
D000818:Animals; D002402:Cathepsin D / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000502:physiology; D049109:Cell Proliferation; D016009:Chi-Square Distribution; D002869:Chromosome Aberrations; D020223:Chromosome Painting; D020732:Cytogenetic Analysis; D004195:Disease Models, Animal; D015180:Electrophoresis, Gel, Two-Dimensional; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology; D006801:Humans; D008297:Male; D051379:Mice; D016513:Mice, SCID; D009368:Neoplasm Transplantation; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D011506:Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism*; D040901:Proteomics; D018709:Statistics, Nonparametric; D001743:Urinary Bladder / Q000378:metabolism / Q000473:pathology; D001749:Urinary Bladder Neoplasms / Q000235:genetics* / Q000378:metabolism*
D011506:Proteins; D002402:Cathepsin D; D046988:Proteasome Endopeptidase Complex