Functional and molecular characterization of the epithelioid to round transition in human colorectal cancer LoVo cells.
In subclones of the human colon cancer LoVo cell line, there is a reproducible spontaneous transition from an epithelioid (E) to a round (R) morphotype. The E to R transition is associated with increased cell growth, absence of E-cadherin-dependent compaction in a slow aggregation assay, loss of contact inhibition of motility and directional migration in a wound filling motility assay. Furthermore, none of the E subclones from LoVo was invasive into chick heart fragments. This is in contrast to the R subclones that were either nonadherent or adherent and invasive. Macroarray analysis demonstrated transcriptional downregulation of plakoglobin in R type LoVo cells and this was confirmed at the level of the mRNA by quantitative RT-PCR. Western blotting showed lower expression of all components of the E-cadherin/catenin complex in R subclones. Interestingly, treatment of R subclones with the demethylating agent 5-aza-2'-deoxycytidine resulted in restoration of the E morphotype, higher expression of E-cadherin, but not plakoglobin mRNA, and higher expression of E-cadherin and plakoglobin at the protein level.
Debruyne
Philip R
PR
Department of Radiotherapy and Nuclear Medicine, Ghent University Hospital, B-9000 Ghent, Belgium.
Vermeulen
Stefan J
SJ
Berx
Geert
G
Pocard
Marc
M
Correia da Rocha
Ana-Sofia
AS
Li
Xuedong
X
Cirnes
Luis
L
Poupon
Marie-France
MF
van Roy
Frans M
FM
Mareel
Marc M
MM
D001374:Azacitidine / Q000031:analogs & derivatives* / Q000633:toxicity; D002449:Cell Aggregation / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods; D002465:Cell Movement / Q000187:drug effects; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D003110:Colonic Neoplasms / Q000235:genetics* / Q000473:pathology*; D000077209:Decitabine; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D014644:Genetic Variation; D006801:Humans; D007413:Intestinal Mucosa / Q000473:pathology*; D012333:RNA, Messenger / Q000235:genetics; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D012333:RNA, Messenger; D000077209:Decitabine; D001374:Azacitidine