Expression and Function of Classical Protein Kinase C Isoenzymes in Gastric Cancer Cell Line and Its Drug-resistant Sublines

Overview

Journal World J Gastroenterol

Publisher Baishideng Publishing Group

Specialty Gastroenterology

Date 2002 Jun 5

PMID 12046066

Citations 14

Authors

Ying Han

Zhe-Yi Han

Xin-Min Zhou

Ru Shi

Yue Zheng

Yong-Quan Shi

Ji-Yan Miao

Bo-Rong Pan

Dai-Ming Fan

Affiliations

Soon will be listed here.

Abstract

Aim: To investigate the expression and function of classical protein kinase C (PKC) isoenzymes in inducing MDR phenotype in gastric cancer cells.

Methods: Two cell lines were used in the study: gastric cancer cell SGC7901 and its drug-resistant cell SGC7901/VCR stepwise-selected by vincristine 0.3, 0.7 and 1.0 mg.L(-1), respectively. The expression of classical PKC (cPKC) isoenzymes in SGC7901 cells and SGC7901/VCR cells were detected using immunofluorescent cytochemistry, laser confocal scanning microscope and Western blot. The effects of anti-PKC isoenzymes antibody on adriamycin accumulation in SGC7901/VCR cells were determined using flow cytometric analysis.

Results: (1)SGC7901 cells exhibited positive staining of PKC-alpha. SGC7901/VCR cells exhibited stronger staining of PKC-alpha than SGC7901 cells. The higher dosage vincristine selected, the much stronger staining of PKC-alpha was observed on SGC7901/VCR cells. (2)Both SGC7901 and SGC7901/VCR cells exhibited positive staining of PKC-beta I and PKC-beta II with no significant difference. (3) Compared with SGC7901, SGC7901/VCR cells had decreased adriamycin accumulation and retention. Accumulation of adriamycin in SGC7901 was 5.21+/-2.56 mg.L(-1),in SGC7901/VCR 0.3 was 0.85+/-0.29 mg.L(-1), in SGC7901/VCR 0.7 was 0.81+/-0.32 mg.L(-1), and in SGC7901/VCR 1.0 was 0.80+/-0.33 mg.L(-1); Retention of adriamycin in SGC 7901 was 2.51+/-1.23 mg.L(-1), in SGC7901/VCR 0.3 was 0.47+/-0.14 mg.L(-1), in SGC7901/VCR 0.7 was 0.44+/-0.15 mg.L(-1), and in SGC 7901/VCR 1.0 was 0.41+/-0.11 mg.L(-1). (4) Fluorescence intensity presented adriamycin accumulation in SGC7901/VCR cells was increased from 1.14+/-0.36 to 2.71+/-0.94 when cells were co-incubated with anti-PKC-alpha but not with anti-PKC-beta I PKC-beta II and PKCgamma antibodies.

Conclusion: PKC-alpha, but not PKC-beta I, PKC-beta II or PKCgamma, may play a role in multidrug resistance of gastric cancer cells SGC7901/VCR.

Citing Articles

Protein Kinase C (PKC) Isozymes as Diagnostic and Prognostic Biomarkers and Therapeutic Targets for Cancer.

Kawano T, Inokuchi J, Eto M, Murata M, Kang J Cancers (Basel). 2022; 14(21).

PMID: 36358843 PMC: 9658272. DOI: 10.3390/cancers14215425.

The inhibitory effect of pseudolaric acid B on gastric cancer and multidrug resistance via Cox-2/PKC-α/P-gp pathway.

Sun Q, Li Y PLoS One. 2014; 9(9):e107830.

PMID: 25250794 PMC: 4176716. DOI: 10.1371/journal.pone.0107830.

Reversal of multidrug resistance in gastric cancer cells by CDX2 downregulation.

Yan L, Wang X, Yang J, Lian C, Kong F, Wei W World J Gastroenterol. 2013; 19(26):4155-65.

PMID: 23864778 PMC: 3710417. DOI: 10.3748/wjg.v19.i26.4155.

Protein kinase Cα protects against multidrug resistance in human colon cancer cells.

Lee S, Shehzad A, Jung J, Sonn J, Lee J, Park J Mol Cells. 2012; 34(1):61-9.

PMID: 22639047 PMC: 3887773. DOI: 10.1007/s10059-012-0087-1.

Caveolin-1 promotes resistance to chemotherapy-induced apoptosis in Ewing's sarcoma cells by modulating PKCalpha phosphorylation.

Tirado O, MacCarthy C, Fatima N, Villar J, Mateo-Lozano S, Notario V Int J Cancer. 2009; 126(2):426-36.

PMID: 19609943 PMC: 2794946. DOI: 10.1002/ijc.24754.

References

Merritt J, Sullivan J, Drew L, Khan A, Wilson K, Mulqueen M . The bisindolylmaleimide protein kinase C inhibitor, Ro 32-2241, reverses multidrug resistance in KB tumour cells. Cancer Chemother Pharmacol. 1999; 43(5):371-8. DOI: 10.1007/s002800050909. View

van Gijn R, van Tellingen O, Haverkate E, Bult A, Beijnen J . Pharmacokinetics and metabolism of the staurosporine analogue CGP 41 251 in mice. Invest New Drugs. 1999; 17(1):29-41. DOI: 10.1023/a:1006260217400. View

Lopez-Soriano F, Argiles J . Lipids: A key role in multidrug resistance? (Review). Int J Oncol. 2000; 16(4):783-98. DOI: 10.3892/ijo.16.4.783. View

Matsumoto Y, Kunishio K, Nagao S . Increased phosphorylation of DNA topoisomerase II in etoposide resistant mutants of human glioma cell line. J Neurooncol. 2000; 45(1):37-46. DOI: 10.1023/a:1006346624083. View

Shtil A, Ktitorova O, Kakpakova E, Holian O . Differential effects of the MDR1 (multidrug resistance) gene-activating agents on protein kinase C: evidence for redundancy of mechanisms of acquired MDR in leukemia cells. Leuk Lymphoma. 2001; 40(1-2):191-5. DOI: 10.3109/10428190009054896. View

Radeff J, Nagy Z, Stern P . Involvement of PKC-beta in PTH, TNF-alpha, and IL-1 beta effects on IL-6 promoter in osteoblastic cells and on PTH-stimulated bone resorption. Exp Cell Res. 2001; 268(2):179-88. DOI: 10.1006/excr.2001.5283. View

Kim M, Lim W, Cha J, An N, Yoo S, Park J . The activation of PI 3-K and PKC zeta in PMA-induced differentiation of HL-60 cells. Cancer Lett. 2001; 171(1):79-85. DOI: 10.1016/s0304-3835(01)00505-5. View

Gill P, Gescher A, Gant T . Regulation of MDR1 promoter activity in human breast carcinoma cells by protein kinase C isozymes alpha and theta. Eur J Biochem. 2001; 268(15):4151-7. DOI: 10.1046/j.1432-1327.2001.02326.x. View

Shindo M, Irie K, Nakahara A, Ohigashi H, Konishi H, Kikkawa U . Toward the identification of selective modulators of protein kinase C (PKC) isozymes: establishment of a binding assay for PKC isozymes using synthetic C1 peptide receptors and identification of the critical residues involved in the phorbol ester.... Bioorg Med Chem. 2001; 9(8):2073-81. DOI: 10.1016/s0968-0896(01)00100-6. View

10.

Lum H, Podolski J, Gurnack M, Schulz I, Huang F, Holian O . Protein phosphatase 2B inhibitor potentiates endothelial PKC activity and barrier dysfunction. Am J Physiol Lung Cell Mol Physiol. 2001; 281(3):L546-55. DOI: 10.1152/ajplung.2001.281.3.L546. View

11.

Boesch D, Garvin J . Age-dependent activation of PKC isoforms by angiotensin II in the proximal nephron. Am J Physiol Regul Integr Comp Physiol. 2001; 281(3):R861-7. DOI: 10.1152/ajpregu.2001.281.3.R861. View

12.

Manier D, Shelton R, Sulser F . Cross-talk between PKA and PKC in human fibroblasts: what are the pharmacotherapeutic implications?. J Affect Disord. 2001; 65(3):275-9. DOI: 10.1016/s0165-0327(00)00278-0. View

13.

Banan A, Fields J, Talmage D, Zhang Y, Keshavarzian A . PKC-beta1 mediates EGF protection of microtubules and barrier of intestinal monolayers against oxidants. Am J Physiol Gastrointest Liver Physiol. 2001; 281(3):G833-47. DOI: 10.1152/ajpgi.2001.281.3.G833. View

14.

Smith J, Yu R, Hinkle P . Activation of MAPK by TRH requires clathrin-dependent endocytosis and PKC but not receptor interaction with beta-arrestin or receptor endocytosis. Mol Endocrinol. 2001; 15(9):1539-48. DOI: 10.1210/mend.15.9.0695. View

15.

Rivedal E, Opsahl H . Role of PKC and MAP kinase in EGF- and TPA-induced connexin43 phosphorylation and inhibition of gap junction intercellular communication in rat liver epithelial cells. Carcinogenesis. 2001; 22(9):1543-50. DOI: 10.1093/carcin/22.9.1543. View

16.

Chang G, Roth C . Structure of MsbA from E. coli: a homolog of the multidrug resistance ATP binding cassette (ABC) transporters. Science. 2001; 293(5536):1793-800. DOI: 10.1126/science.293.5536.1793. View

17.

Meng L, Zhang J, Ding J . Salvicine, a novel DNA topoisomerase II inhibitor, exerting its effects by trapping enzyme-DNA cleavage complexes. Biochem Pharmacol. 2001; 62(6):733-41. DOI: 10.1016/s0006-2952(01)00732-8. View

18.

Mansson E, Paul A, Lofgren C, Ullberg K, Paul C, Eriksson S . Cross-resistance to cytosine arabinoside in a multidrug-resistant human promyelocytic cell line selected for resistance to doxorubicin: implications for combination chemotherapy. Br J Haematol. 2001; 114(3):557-65. DOI: 10.1046/j.1365-2141.2001.02979.x. View

19.

Kumar A, Hawkins K, Hannan M, Ganz M . Activation of PKC-beta(I) in glomerular mesangial cells is associated with specific NF-kappaB subunit translocation. Am J Physiol Renal Physiol. 2001; 281(4):F613-9. DOI: 10.1152/ajprenal.2001.281.4.F613. View

20.

Chen L, Hahn H, Wu G, Chen C, Liron T, Schechtman D . Opposing cardioprotective actions and parallel hypertrophic effects of delta PKC and epsilon PKC. Proc Natl Acad Sci U S A. 2001; 98(20):11114-9. PMC: 58692. DOI: 10.1073/pnas.191369098. View