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. 2000 Nov;83(9):1209–1215. doi: 10.1054/bjoc.2000.1436

Downregulation of intracellular nm23-H1 prevents cisplatin-induced DNA damage in oesophageal cancer cells: possible association with Na+, K+-ATPase

N lizuka 1, K Miyamoto 1,3, A Tangoku 2, H Hayashi 2, S Hazama 2, S Yoshino 2, K Yoshimura 2, K Hirose 4, H Yoshida 5, M Oka 2
PMCID: PMC2363580  PMID: 11027435

Abstract

Previously, we showed that expression of nm23-H1 is associated inversely with sensitivity to cisplatin in human oesophageal squamous cell carcinoma (OSCC). The present study was undertaken to investigate the association of nm23-H1 expression with cisplatin-induced DNA damage in OSCC using antisense nm23-H1 transfectants. YES-2/AS-12, an antisense nm23-H1-transfected OSCC cell line, showed significantly reduced expression of intracellular nm23-H1 protein compared with that in parental YES-2 cells and YES-2/Neo transfectants. Surface expression of nm23-H1 protein was not observed in any of the three cell lines. PCR analysis for DNA damage demonstrated that YES-2/AS-12 cells were more resistant to nuclear and mitochondrial DNA damage by cisplatin than were YES-2/Neo cells. In addition, mitochondrial membrane potentials and DNA fragmentation assays confirmed that YES-2/AS-12 was more resistant than YES-2/Neo to apoptosis induced by cisplatin. In contrast, YES-2/AS-12 was more sensitive to ouabain, a selective inhibitor of Na+, K+-ATPase, than YES-2 and YES-2/Neo. Pre-treatment with ouabain resulted in no differences in cisplatin sensitivity between the three cell lines examined. Intracellular platinum level in YES-2/AS-12 was significantly lower than that in YES-2 and YES-2/Neo following incubation with cisplatin, whereas ouabain pre-treatment resulted in no differences in intracellular platinum accumulations between the three cell lines. Our data support the conclusion that reduced expression of intracellular nm23-H1 in OSCC cells is associated with cisplatin resistance via the prevention of both nuclear and mitochondrial DNA damage and suggest that it may be related to Na+, K+-ATPase activity, which is responsible for intracellular cisplatin accumulation. © 2000 Cancer Research Campaign

Keywords: nm23, cisplatin, DNA damage, mitochondrial membrane potential, oesophageal squamous cell carcinoma

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Selected References

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  1. Andrews P. A., Mann S. C., Huynh H. H., Albright K. D. Role of the Na+, K(+)-adenosine triphosphatase in the accumulation of cis-diamminedichloroplatinum(II) in human ovarian carcinoma cells. Cancer Res. 1991 Jul 15;51(14):3677–3681. [PubMed] [Google Scholar]
  2. Blok L. J., Chang G. T., Steenbeek-Slotboom M., van Weerden W. M., Swarts H. G., De Pont J. J., van Steenbrugge G. J., Brinkmann A. O. Regulation of expression of Na+,K+-ATPase in androgen-dependent and androgen-independent prostate cancer. Br J Cancer. 1999 Sep;81(1):28–36. doi: 10.1038/sj.bjc.6690647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bosset J. F., Gignoux M., Triboulet J. P., Tiret E., Mantion G., Elias D., Lozach P., Ollier J. C., Pavy J. J., Mercier M. Chemoradiotherapy followed by surgery compared with surgery alone in squamous-cell cancer of the esophagus. N Engl J Med. 1997 Jul 17;337(3):161–167. doi: 10.1056/NEJM199707173370304. [DOI] [PubMed] [Google Scholar]
  4. Daoud S. S., Clements M. K., Small C. L. Polymerase chain reaction analysis of cisplatin-induced mitochondrial DNA damage in human ovarian carcinoma cells. Anticancer Drugs. 1995 Jun;6(3):405–412. doi: 10.1097/00001813-199506000-00007. [DOI] [PubMed] [Google Scholar]
  5. Eguchi Y., Srinivasan A., Tomaselli K. J., Shimizu S., Tsujimoto Y. ATP-dependent steps in apoptotic signal transduction. Cancer Res. 1999 May 1;59(9):2174–2181. [PubMed] [Google Scholar]
  6. Ferguson A. W., Flatow U., MacDonald N. J., Larminat F., Bohr V. A., Steeg P. S. Increased sensitivity to cisplatin by nm23-transfected tumor cell lines. Cancer Res. 1996 Jul 1;56(13):2931–2935. [PubMed] [Google Scholar]
  7. Freije J. M., Lawrence J. A., Hollingshead M. G., De la Rosa A., Narayanan V., Grever M., Sausville E. A., Paull K., Steeg P. S. Identification of compounds with preferential inhibitory activity against low-Nm23-expressing human breast carcinoma and melanoma cell lines. Nat Med. 1997 Apr;3(4):395–401. doi: 10.1038/nm0497-395. [DOI] [PubMed] [Google Scholar]
  8. Fulda S., Lutz W., Schwab M., Debatin K. M. MycN sensitizes neuroblastoma cells for drug-induced apoptosis. Oncogene. 1999 Feb 18;18(7):1479–1486. doi: 10.1038/sj.onc.1202435. [DOI] [PubMed] [Google Scholar]
  9. Hazama S., Noma T., Wang F., Iizuka N., Ogura Y., Yoshimura K., Inoguchi E., Hakozaki M., Hirose K., Suzuki T. Tumour cells engineered to secrete interleukin-15 augment anti-tumour immune responses in vivo. Br J Cancer. 1999 Jul;80(9):1420–1426. doi: 10.1038/sj.bjc.6690538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Higashiyama M., Doi O., Yokouchi H., Kodama K., Nakamori S., Tateishi R., Kimura N. Immunohistochemical analysis of nm23 gene product/NDP kinase expression in pulmonary adenocarcinoma: lack of prognostic value. Br J Cancer. 1992 Sep;66(3):533–536. doi: 10.1038/bjc.1992.308. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hishikawa Y., Abe S., Kinugasa S., Yoshimura H., Monden N., Igarashi M., Tachibana M., Nagasue N. Overexpression of metallothionein correlates with chemoresistance to cisplatin and prognosis in esophageal cancer. Oncology. 1997 Jul-Aug;54(4):342–347. doi: 10.1159/000227714. [DOI] [PubMed] [Google Scholar]
  12. Iizuka N., Hirose K., Noma T., Hazama S., Tangoku A., Hayashi H., Abe T., Yamamoto K., Oka M. The nm23-H1 gene as a predictor of sensitivity to chemotherapeutic agents in oesophageal squamous cell carcinoma. Br J Cancer. 1999 Oct;81(3):469–475. doi: 10.1038/sj.bjc.6690717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Iizuka N., Miyamoto K., Okita K., Tangoku A., Hayashi H., Yosino S., Abe T., Morioka T., Hazama S., Oka M. Inhibitory effect of Coptidis Rhizoma and berberine on the proliferation of human esophageal cancer cell lines. Cancer Lett. 2000 Jan 1;148(1):19–25. doi: 10.1016/s0304-3835(99)00264-5. [DOI] [PubMed] [Google Scholar]
  14. Iizuka N., Oka M., Noma T., Nakazawa A., Hirose K., Suzuki T. NM23-H1 and NM23-H2 messenger RNA abundance in human hepatocellular carcinoma. Cancer Res. 1995 Feb 1;55(3):652–657. [PubMed] [Google Scholar]
  15. Iizuka N., Tangoku A., Hayashi H., Yosino S., Abe T., Morioka T., Oka M. The association between nm23-H1 expression and survival in patients with esophageal squamous cell carcinoma. Cancer Lett. 1999 Apr 26;138(1-2):139–144. doi: 10.1016/s0304-3835(98)00387-5. [DOI] [PubMed] [Google Scholar]
  16. Ishikawa N., Shimada N., Munakata Y., Watanabe K., Kimura N. Isolation and characterization of a gene encoding rat nucleoside diphosphate kinase. J Biol Chem. 1992 Jul 15;267(20):14366–14372. [PubMed] [Google Scholar]
  17. Kalinowski D. P., Illenye S., Van Houten B. Analysis of DNA damage and repair in murine leukemia L1210 cells using a quantitative polymerase chain reaction assay. Nucleic Acids Res. 1992 Jul 11;20(13):3485–3494. doi: 10.1093/nar/20.13.3485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kim C. N., Wang X., Huang Y., Ibrado A. M., Liu L., Fang G., Bhalla K. Overexpression of Bcl-X(L) inhibits Ara-C-induced mitochondrial loss of cytochrome c and other perturbations that activate the molecular cascade of apoptosis. Cancer Res. 1997 Aug 1;57(15):3115–3120. [PubMed] [Google Scholar]
  19. Kok T. C., Van der Gaast A., Dees J., Eykenboom W. M., Van Overhagen H., Stoter G., Tilanus H. W., Splinter T. A. Cisplatin and etoposide in oesophageal cancer: a phase II study. Rotterdam Oesophageal Tumour Study Group. Br J Cancer. 1996 Sep;74(6):980–984. doi: 10.1038/bjc.1996.469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kroemer G., Dallaporta B., Resche-Rigon M. The mitochondrial death/life regulator in apoptosis and necrosis. Annu Rev Physiol. 1998;60:619–642. doi: 10.1146/annurev.physiol.60.1.619. [DOI] [PubMed] [Google Scholar]
  21. Leone A., Flatow U., VanHoutte K., Steeg P. S. Transfection of human nm23-H1 into the human MDA-MB-435 breast carcinoma cell line: effects on tumor metastatic potential, colonization and enzymatic activity. Oncogene. 1993 Sep;8(9):2325–2333. [PubMed] [Google Scholar]
  22. Lindmark G. NM-23 H1 immunohistochemistry is not useful as predictor of metastatic potential of colorectal cancer. Br J Cancer. 1996 Nov;74(9):1413–1418. doi: 10.1038/bjc.1996.557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mann S. C., Andrews P. A., Howell S. B. Modulation of cis-diamminedichloroplatinum(II) accumulation and sensitivity by forskolin and 3-isobutyl-1-methylxanthine in sensitive and resistant human ovarian carcinoma cells. Int J Cancer. 1991 Jul 30;48(6):866–872. doi: 10.1002/ijc.2910480613. [DOI] [PubMed] [Google Scholar]
  24. Marshall L. J., Muimo R., Riemen C. E., Mehta A. Na+ and K+ regulate the phosphorylation state of nucleoside diphosphate kinase in human airway epithelium. Am J Physiol. 1999 Jan;276(1 Pt 1):C109–C119. doi: 10.1152/ajpcell.1999.276.1.C109. [DOI] [PubMed] [Google Scholar]
  25. O'Neill C. F., Koberle B., Masters J. R., Kelland L. R. Gene-specific repair of Pt/DNA lesions and induction of apoptosis by the oral platinum drug JM216 in three human ovarian carcinoma cell lines sensitive and resistant to cisplatin. Br J Cancer. 1999 Dec;81(8):1294–1303. doi: 10.1038/sj.bjc.6694381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ohmori T., Nishio K., Ohta S., Kubota N., Adachi M., Komiya K., Saijo N. Ouabain-resistant non-small-cell lung-cancer cell line shows collateral sensitivity to cis-diamminedichloroplatinum(II) (CDDP). Int J Cancer. 1994 Apr 1;57(1):111–116. doi: 10.1002/ijc.2910570120. [DOI] [PubMed] [Google Scholar]
  27. Oka M., Hirazawa K., Yamamoto K., Iizuka N., Hazama S., Suzuki T., Kobayashi N. Induction of Fas-mediated apoptosis on circulating lymphocytes by surgical stress. Ann Surg. 1996 Apr;223(4):434–440. doi: 10.1097/00000658-199604000-00013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Oka M., Iizuka N., Yamamoto K., Gondo T., Abe T., Hazama S., Akitomi Y., Koishihara Y., Ohsugi Y., Ooba Y. The influence of interleukin-6 on the growth of human esophageal cancer cell lines. J Interferon Cytokine Res. 1996 Dec;16(12):1001–1006. doi: 10.1089/jir.1996.16.1001. [DOI] [PubMed] [Google Scholar]
  29. Olivero O. A., Chang P. K., Lopez-Larraza D. M., Semino-Mora M. C., Poirier M. C. Preferential formation and decreased removal of cisplatin-DNA adducts in Chinese hamster ovary cell mitochondrial DNA as compared to nuclear DNA. Mutat Res. 1997 Jun 13;391(1-2):79–86. doi: 10.1016/s0165-1218(97)00037-2. [DOI] [PubMed] [Google Scholar]
  30. Olivero O. A., Semino C., Kassim A., Lopez-Larraza D. M., Poirier M. C. Preferential binding of cisplatin to mitochondrial DNA of Chinese hamster ovary cells. Mutat Res. 1995 Apr;346(4):221–230. doi: 10.1016/0165-7992(95)90039-x. [DOI] [PubMed] [Google Scholar]
  31. Otero A. S., Doyle M. B., Hartsough M. T., Steeg P. S. Wild-type NM23-H1, but not its S120 mutants, suppresses desensitization of muscarinic potassium current. Biochim Biophys Acta. 1999 Mar 8;1449(2):157–168. doi: 10.1016/s0167-4889(99)00009-9. [DOI] [PubMed] [Google Scholar]
  32. Russell R. L., Pedersen A. N., Kantor J., Geisinger K., Long R., Zbieranski N., Townsend A., Shelton B., Brünner N., Kute T. E. Relationship of nm23 to proteolytic factors, proliferation and motility in breast cancer tissues and cell lines. Br J Cancer. 1998 Sep;78(6):710–717. doi: 10.1038/bjc.1998.566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  34. Scambia G., Ferrandina G., Marone M., Benedetti Panici P., Giannitelli C., Piantelli M., Leone A., Mancuso S. nm23 in ovarian cancer: correlation with clinical outcome and other clinicopathologic and biochemical prognostic parameters. J Clin Oncol. 1996 Feb;14(2):334–342. doi: 10.1200/JCO.1996.14.2.334. [DOI] [PubMed] [Google Scholar]
  35. Shimada M., Taguchi K., Hasegawa H., Gion T., Shirabe K., Tsuneyoshi M., Sugimachi K. Nm23-H1 expression in intrahepatic or extrahepatic metastases of hepatocellular carcinoma. Liver. 1998 Oct;18(5):337–342. doi: 10.1111/j.1600-0676.1998.tb00815.x. [DOI] [PubMed] [Google Scholar]
  36. Stahl J. A., Leone A., Rosengard A. M., Porter L., King C. R., Steeg P. S. Identification of a second human nm23 gene, nm23-H2. Cancer Res. 1991 Jan 1;51(1):445–449. [PubMed] [Google Scholar]
  37. Steeg P. S., Bevilacqua G., Kopper L., Thorgeirsson U. P., Talmadge J. E., Liotta L. A., Sobel M. E. Evidence for a novel gene associated with low tumor metastatic potential. J Natl Cancer Inst. 1988 Apr 6;80(3):200–204. doi: 10.1093/jnci/80.3.200. [DOI] [PubMed] [Google Scholar]
  38. Timmer-Bosscha H., Timmer A., Meijer C., de Vries E. G., de Jong B., Oosterhuis J. W., Mulder N. H. cis-diamminedichloroplatinum(ii) resistance in vitro and in vivo in human embryonal carcinoma cells. Cancer Res. 1993 Dec 1;53(23):5707–5713. [PubMed] [Google Scholar]
  39. Tokunaga Y., Urano T., Furukawa K., Kondo H., Kanematsu T., Shiku H. Reduced expression of nm23-H1, but not of nm23-H2, is concordant with the frequency of lymph-node metastasis of human breast cancer. Int J Cancer. 1993 Aug 19;55(1):66–71. doi: 10.1002/ijc.2910550113. [DOI] [PubMed] [Google Scholar]
  40. Urano T., Furukawa K., Shiku H. Expression of nm23/NDP kinase proteins on the cell surface. Oncogene. 1993 May;8(5):1371–1376. [PubMed] [Google Scholar]
  41. Willems R., Van Bockstaele D. R., Lardon F., Lenjou M., Nijs G., Snoeck H. W., Berneman Z. N., Slegers H. Decrease in nucleoside diphosphate kinase (NDPK/nm23) expression during hematopoietic maturation. J Biol Chem. 1998 May 29;273(22):13663–13668. doi: 10.1074/jbc.273.22.13663. [DOI] [PubMed] [Google Scholar]
  42. Yoon K. L., Modica-Napolitano J. S., Ernst S. G., Aprille J. R. Denaturing gradient gel method for mapping single base changes in human mitochondrial DNA. Anal Biochem. 1991 Aug 1;196(2):427–432. doi: 10.1016/0003-2697(91)90489-g. [DOI] [PubMed] [Google Scholar]
  43. Zaborina O., Li X., Cheng G., Kapatral V., Chakrabarty A. M. Secretion of ATP-utilizing enzymes, nucleoside diphosphate kinase and ATPase, by Mycobacterium bovis BCG: sequestration of ATP from macrophage P2Z receptors? Mol Microbiol. 1999 Mar;31(5):1333–1343. doi: 10.1046/j.1365-2958.1999.01240.x. [DOI] [PubMed] [Google Scholar]

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