A study of membrane activity in rat prostate cancer cells: an evaluation of the FM1-43 dye technique.
A study was initiated to test whether the FM1-43 dye technique could be applied to the study of endocytic membrane activity in two rodent prostate cancer (MAT-LyLu and AT-2) cell lines of markedly different metastatic ability. The lipophilic dye FM1-43, which has frequently been used to monitor endo/exocytic activity in excitable cells was employed. We found, as in excitable tissues, that both strongly metastatic (MAT-LyLu) and weakly metastatic (AT-2) cells in culture take up FM1-43 to give vesicular staining of a variable pattern, which appeared to differ between the two cell lines. However, unlike excitable tissues, neither cell line subsequently released the dye. Indeed, both cell lines retained the dye through several rounds of cell division suggesting that dye incorporated by cells does not enter the endo/exocytotic cycle. Uptake of dye was independent of temperature, Na+/K+ gradients, pH or metabolism. We suggest that passive accumulation of FM1-43 can occur in cancer cells and should not, automatically, be interpreted as evidence of endocytosis.
Mycielska
M E
ME
Department of Biology, Imperial College of Science, Technology and Medicine, London, UK.
Schäfer
M
M
Fraser
S P
SP
Djamgoz
M B
MB
Bashford
C L
CL
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D002470:Cell Survival; D004705:Endocytosis; D005089:Exocytosis; D005456:Fluorescent Dyes / Q000493:pharmacokinetics / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007473:Ion Channels / Q000378:metabolism; D008297:Male; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D011188:Potassium / Q000378:metabolism; D011471:Prostatic Neoplasms / Q000378:metabolism*; D011726:Pyridinium Compounds / Q000493:pharmacokinetics / Q000494:pharmacology; D000644:Quaternary Ammonium Compounds / Q000493:pharmacokinetics / Q000494:pharmacology; D051381:Rats; D012964:Sodium / Q000378:metabolism; D013696:Temperature; D014407:Tumor Cells, Cultured
C073804:FM1 43; D005456:Fluorescent Dyes; D007473:Ion Channels; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds; D012964:Sodium; D011188:Potassium