Analysis of calcium-dependent protein kinase-C isoenzymes in intrinsically resistant cloned lines of LoVo cells: reversal of resistance by kinase inhibitor 1-(5-isoquinolinylsulfonyl) 2-methylpiperazine.
In order to investigate the involvement of Protein Kinase C (PKC) in the signal transduction mechanisms related to intrinsic chemoresistance, two cellular clones were isolated from LoVo/WT colon adenocarcinoma cell line and their cytogenetic pattern was studied: LoVo C1.7 was intrinsically resistant to Doxorubicin while LoVo C1.5 showed the same resistance index as the mixed parental cell population. Two PKC isoforms, immunologically identified as beta and alpha PKC, were isolated from the cytosolic fraction of all cell types and one single peak of alpha PKC was obtained from the particulate fraction. Resistant LoVo C1.7 cells showed a significant increase of PKC activity; preincubation with H-7 induced PKC inhibition and reversal of drug resistance. These data suggest that in our cell system the identified calcium-dependent PKC subtypes can play a role in the mechanisms of intrinsic resistance.
University of Milan, Institute of Pharmacology, Italy.
E
E
Dolfini
T
T
Dasdia
G
G
Perletti
M
M
Romagnoni
F
F
Piccinini
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000230:Adenocarcinoma; D002118:Calcium / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D002845:Chromatography; D002869:Chromosome Aberrations; D003110:Colonic Neoplasms; D003600:Cytosol / Q000201:enzymology; D004317:Doxorubicin / Q000633:toxicity*; D004351:Drug Resistance; D017886:Durapatite; D006801:Humans; D006882:Hydroxyapatites; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D007546:Isoquinolines / Q000494:pharmacology*; D007621:Karyotyping; D010879:Piperazines / Q000494:pharmacology*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D014410:Tumor Stem Cell Assay
D006882:Hydroxyapatites; D007527:Isoenzymes; D007546:Isoquinolines; D010879:Piperazines; D004317:Doxorubicin; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D017886:Durapatite; D011493:Protein Kinase C; D002118:Calcium