Protein kinase C and casein kinase II activities in two human colon carcinoma cell lines, HT-29 and CaCo-2: Possible correlation with differentiation

1990 ◽  
Vol 10 (3) ◽  
pp. 293-299 ◽  
Author(s):  
Ewa Rydell ◽  
Karl-Eric Magnusson ◽  
Anita Sjö ◽  
Krister Axelsson
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Colon Carcinoma ◽  
Carcinoma Cell ◽  
Human Colon ◽  
Human Colon Carcinoma Cell ◽  
Carcinoma Cell Lines ◽  
Kinase C ◽  
Human Colon Carcinoma ◽  
Ht 29

Protein kinase C (PK-C) and casein kinase II (CK-II) activities were studied in two human colon carcinoma cell lines (HT-29 and CaCO-2) undergoing differentiation in vitro resulting, in small-intestine-like cells. CaCo-2 cells, when grown under standard conditions, appear to undergo spontaneous differentiation. In these cells PK-C and CK-II activities were determined on day 5, 10 and 15. No significant differences in activities were seen either in PK-C or CK-II activity. HT-29 cells, when grown in glucose-free medium can be stimulated to undergo differentiation which is completed within 20 days. PK-C and CK-II activities were determined after 5, 10, 15, 20 and 25 days, respectively. PK-C activity rose from 7.9±3.5 pmole32P/mg protein/min at day 5 to 37.5±14.8 pmole32P/mg protein/min at day 20. After 25 days the activity was reduced to 20.0±7.8 pmole32P/mg protein/min. CK-II activity did not change significantly during day 5 to 20, but on day 25 there was a significant decrease in CK-II activity from 94.9±6.4 pmole32P/mg protein/min (day 20) to 62.6±3.9 pmole32P/mg protein/min (day 25) p=0.003. The results in this study indicate a role for PK-C and CK-II in cell growth and differentiation.

scholarly journals Expression and protein kinase C-dependent regulation of peptide/H+ co-transport system in the Caco-2 human colon carcinoma cell line

1994 ◽  
Vol 299 (1) ◽  
pp. 253-260 ◽  
Author(s):  
M Brandsch ◽  
Y Miyamoto ◽  
V Ganapathy ◽  
F H Leibach
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Cell Line ◽  
Transport System ◽  
Carcinoma Cell ◽  
Human Colon ◽  
Human Colon Carcinoma Cell ◽  
Colon Carcinoma Cell Line ◽  
Kinase C ◽  
Human Colon Carcinoma

The characteristics of the transport of the dipeptide glycylsarcosine were studied in the human colon carcinoma cell line Caco-2 grown as a monolayer on impermeable plastic support. Transport of glycylsarcosine in these cells was found to be Na(+)-independent, but was stimulated by an inwardly directed H+ gradient. This H(+)-dependent transport of glycylsarcosine was inhibited by di- and tri-peptides and also by the beta-lactam antibiotic cephalexin, but was unaffected by the amino acids glycine and leucine. The transport system exhibited a Michaelis-Menten constant (Kt) of 1.1 +/- 0.1 mM for glycylsarcosine. The specific activity of the transport system in this cell line was found to be maximal when the cultures were confluent. Treatment of the cells with phorbol esters which activate protein kinase C resulted in a significant inhibition of the transport system. This inhibition was specific and could be blocked if treatment was done in the presence of staurosporine, an inhibitor of protein kinase C. Kinetic analysis revealed that the inhibition was associated with a decrease in the maximal velocity, the Kt remaining unaffected. The phorbol-ester-induced inhibition of the peptide-transport system was not prevented by co-treatment with cycloheximide, an inhibitor of cellular protein synthesis. In addition, there was no change in the intracellular pH following treatment with the phorbol ester, suggesting that the effect was not due to alterations in the transmembrane pH gradient. It is concluded that the peptide/H+ co-transport system, which is known to exist in the normal intestine, is expressed in Caco-2 cells and that the function of the transport system is under the regulatory control of protein kinase C.

Regulation of taurine transport in human colon carcinoma cell lines (HT-29 and Caco-2) by protein kinase C

1993 ◽  
Vol 264 (5) ◽  
pp. G939-G946 ◽  
Author(s):  
M. Brandsch ◽  
Y. Miyamoto ◽  
V. Ganapathy ◽  
F. H. Leibach
Keyword(s):  
Protein Kinase C ◽  
Cell Line ◽  
Carcinoma Cell ◽  
Human Colon ◽  
Human Colon Carcinoma Cell ◽  
Taurine Transport ◽  
Kinase C ◽  
Human Colon Carcinoma ◽  
Taurine Uptake ◽  
Ht 29

The effect of phorbol 12-myristate 13-acetate (PMA), a phorbol ester known to stimulate protein kinase C, on taurine transport was studied in the human colon carcinoma cell line HT-29. PMA (1 microM) was found to inhibit taurine uptake in confluent monolayers of this cell line by approximately 70% after pretreatment of the cells with the compound for 1 h (IC50 = 42.7 +/- 2.6 nM). The inhibitory effect of PMA on the taurine transporter could be confirmed by using beta-alanine, another substrate for the transporter. Kinetic analysis of taurine uptake indicated that the PMA effect was associated with a decrease in the maximal velocity (954 +/- 26 vs. 676 +/- 28 pmol.10 min-1.mg of protein-1) and an increase in the Michaelis-Menten constant (9.8 +/- 0.5 vs. 13.3 +/- 1.0 microM). The inhibition of taurine uptake could be blocked by cotreatment of the cultures with staurosporine, an inhibitor of protein kinase C. The inactive phorbol ester 4 alpha-phorbol 12,13-didecanoate had no effect. Treatment of the cells with PMA did not alter the uptake of leucine and lysine, stimulated the uptake of aspartic acid, and inhibited the uptake of proline. The PMA effect on taurine uptake was not prevented by cycloheximide, actinomycin D, colchicine, or cytochalasin D. Comparison of the taurine transport activity in HT-29 cells with that in Caco-2, another human colon carcinoma cell line, revealed that the latter cell line also expressed the taurine transporter but at a much reduced level (about one-fifth compared with HT-29).(ABSTRACT TRUNCATED AT 250 WORDS)

Modulation of EGF Receptor Expression by Differentiating Agents in Human Colon Carcinoma Cell Lines

1990 ◽  
Vol 2 (10) ◽  
pp. 345-355 ◽  
Author(s):  
Lizabeth Deutsch Murphy ◽  
Eva M. Valverius ◽  
Maria Tsokos ◽  
Lyn A. Mickley ◽  
Neal Rosen ◽  
...  
Keyword(s):  
Colon Carcinoma ◽  
Cell Lines ◽  
Egf Receptor ◽  
Carcinoma Cell ◽  
Human Colon ◽  
Receptor Expression ◽  
Human Colon Carcinoma Cell ◽  
Carcinoma Cell Lines ◽  
Human Colon Carcinoma ◽  
Differentiating Agents

Activities of purine-metabolizing enzymes in human colon carcinoma cell lines and xenograft tumors

1981 ◽  
Vol 30 (7) ◽  
pp. 793-798 ◽  
Author(s):  
Gerald W. Crabtree ◽  
Daniel L. Dexter ◽  
Johanna D. Stoeckler ◽  
Todd M. Savarese ◽  
Lucy Y. Ghoda ◽  
...  
Keyword(s):  
Colon Carcinoma ◽  
Cell Lines ◽  
Carcinoma Cell ◽  
Human Colon ◽  
Human Colon Carcinoma Cell ◽  
Metabolizing Enzymes ◽  
Carcinoma Cell Lines ◽  
Human Colon Carcinoma

scholarly journals Characterization of human colon carcinoma cell lines isolated from a single primary tumour

1983 ◽  
Vol 47 (3) ◽  
pp. 373-381 ◽  
Author(s):  
M G Brattain ◽  
M E Marks ◽  
J McCombs ◽  
W Finely ◽  
D E Brattain
Keyword(s):  
Colon Carcinoma ◽  
Cell Lines ◽  
Carcinoma Cell ◽  
Primary Tumour ◽  
Human Colon ◽  
Human Colon Carcinoma Cell ◽  
Carcinoma Cell Lines ◽  
Human Colon Carcinoma
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