New method for separation of subpopulations from a heterogeneous colon cancer cell line.
BACKGROUND
To understand the heterogeneity of human colon cancers, a new method to separate cancer subpopulations was developed.


METHODS
Cells from a human colon cancer cell line, DLD-1, were seeded on an 8 microm pore membrane. After six hours, the cells which remained beneath the membrane as well as the cells which dropped onto the 24-well plate were collected. To clarify the differences between the two subpopulations, transepithelial electrical resistance (TEER) and immunocytochemistry were evaluated.


RESULTS
Two subpopulations, clones D and A, were separated from DLD-1 with the newly developed method. Both subpopulations showed quite different TEER values and different arrangements of cell-cell contact. In addition, the distinct subcellular localizations of claudin family proteins and zonula occludens-1 (ZO-1) were identified.


CONCLUSIONS
A new separation method to isolate colon cancer subpopulations was established in which the intercellular junctions differed. This method can be considered as a helpful tool in the investigation of colon cancer heterogeneity.
Takaya
Yoshihiro
Y
Department of Pharmacology, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan.
Tanaka
Kunihiko
K
Hirakawa
Chiasa
C
Tagawa
Yutaka
Y
Niwa
Masami
M
D018938:Caco-2 Cells; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000379:methods*; D003110:Colonic Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D017097:Electric Impedance; D006801:Humans; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D019108:Tight Junctions / Q000235:genetics / Q000378:metabolism
D008565:Membrane Proteins; D012333:RNA, Messenger