Abstract
We examined chemosensitivity to 5-fluorouracil (5-FU) in four human gastric cancer cell lines, by analyzing the expression of p53 and its related genes. Treatment with 1mM 5-FU induced variable degrees of apoptosis in the cultured cells. The apoptotic indices 72 h after treatment were approximately 14% in MKN-74 (wild-type p53 gene), 12% in MKN-45 (wild-type), 3% in MKN-28 (mutated) and 0.5% in KATO-III cells (deleted), respectively. On the other hand, 50 μM 5-FU had little effect on the induction of apoptosis in MKN-74 cells, the value being approximately 2% after 72 h. Induction of P53 expression was noted 3 h after initiating the treatment, followed by the induction of P21/Waf1 after 6 h in both MKN-74 and MKN-45 cells. The same expression mode was noted in MKN-74 treated with 50 μM 5-FU. Conversely, the level of P53 expression was constant in MKN-28 cells and absent in KATO-III cells, in which P21/Waf1 had never been induced. The Bax/Bcl-2 expression ratio was gradually elevated for up to 72 h in MKN-74 and MKN-45 cells treated with 1mM 5-FU; in contrast, it was unchanged in MKN-28 and KATO-III cells, and MKN-74 treated with 50 μM 5-FU. These results might indicate that (1) 1mM 5-FU induces apoptosis in cultured gastric cancer cells carrying the wild-type p53 gene, but not those carrying the mutated type or a gene deletion, and (2) the elevated Bax/Bcl-2 expression ratio plays a more crucial role than the higher expression of P21/Waf1 in the induction of p53- gene dependent apoptosis.
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References
Parker WB, Cheng YC. Metabolism and mechanism of action of 5-fluorouracil. Pharmacol Ther 1990; 48: 381–395.
Nabeya Y, Loganzo F Jr, Maslak P, et al. The mutational status of p53 protein in gastric and esophageal adenocarcinoma cell lines predicts sensitivity to chemotherapeutic agents. Int J Cancer 1995; 64: 37–46.
Pickard M, Dive C, Kinsella AR. Differences in resistance to 5-fluorouracil as a function of cell cycle delay and not apoptosis. Br J Cancer 1995; 72: 1389–1396.
Yonish-Rouach E, Resnitzky D, Lotem J, Sachs L, Kimchi A, Oren M. Wild-type p53 induces apoptosis of myeloid leukaemic cells that is inhibited by Interleukin-6. Nature 1991; 352: 345–347.
Shaw P, Bovey R, Tardy S, Sahli R, Sordat B, Costa J. Induction of apoptosis by wild-type p53 in a human colon tumor-derived cell line. Proc Natl Acad Sci USA 1992; 89: 4495–4499.
Ramqvist T, Magnusson KP, Wang Y, Szekely L, Klein G, Wiman KG. Wild-type p53 induces apoptosis in a Burkitt lymphoma (BL) line that carries mutant p53. Oncogene 1993; 8: 1495–1500.
Ryan JJ, Danish R, Gottlieb CA, Clarke MF. Cell cycle analysis of p53-induced cell death in murine erythroleukemia cells. Mol Cell Biol 1993; 13: 711–719.
Wang Y, Ramqvist T, Szekely L, Axelson H, Klein G, Wiman KG. Reconstitution of wild-type p53 expression triggers apoptosis in a p53-negative v-myc retrovirus-induced T-cell lymphoma line. Cell Growth Differ 1993; 4: 467–473.
Eizenberg O, Faber-Elmanr A, Gottlieb E, Oren M, Rotter V, Schwartz M. Direct involvement of p53 in programmed cell death of oligodendrocytes EMBO J 1995; 14: 1136–1144.
Kastsan MB, Onyekwere O, Sidransky D, Vogelstein B, Craig RW. Participation of p53 protein in the cellular response to DNA damage. Cancer Res 1991; 51: 6304–6311.
Fritsche M, Haessler C, Brandner G. Induction of nuclear accumulation of the tumor-suppressor protein p53 by DNA-damaging agents. Oncogene 1993; 8: 307–318.
Nelson WG, Kastan MB. DNA strand breaks: the DNA template alterations that trigger p53-dependent DNA damage response pathways. Mol Cell Biol 1994; 14: 1815–1823.
Hall PA, Meek D, Lane DP. p53-Integrating the complexity. J Pathol 1996; 180: 1–5.
El-Deiry WS, Harper JW, O'Connor PM, et al. WAF1/CIP1 is induced in p53-mediated G1 arrest and apoptosis. Cancer Res 1994; 54: 1169–1174.
Harper JW, Adami GR, Wei N, Keyomarsi K, Elledge SJ. The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases. Cell 1993; 75: 805–816.
Waldman T, Kinzler KW, Vogelstein B. p21 is necessary for the p53-mediated G1 arrest in human cancer cells. Cancer Res 1995; 55: 5187–5190.
Miyashita T, Reed JC. Tumor suppressor p53 is a direct transcriptional activator of the human bax gene. Cell 1995; 80: 293–299.
Oltvai ZN, Milliman CL, Korsmeyer SJ. Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programed cell death. Cell 1993; 74: 609–619.
Ishida M, Gomyo Y, Tatebe S, Ohfuji S, Ito H. Apoptosis in human gastric mucosa, chronic gastritis, dysplasia and carcinoma: analysis by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling. Virchows Arch 1996; 428: 229–235.
Sugamura K, Makino M, Shirai H, et al. Enhanced induction of apoptosis of human gastric carcinoma cells after preoperative treatment with 5-fluorouracil. Cancer 1997; 79: 12–17.
Makino M, Shirai H, Sugamura K, et al. Increased induction of apoptosis of human colorectal cancer cells after preoperative treatment with 5-fluorouracil. Oncol Rep 1996; 3: 281–285
Hojo H. Establishment of cultured cell lines of human stomach cancer origin and their morphological characteristics. Niigata Igakukai Zassi 1977; 91: 737–752.
Sekiguchi M, Sakakibara K, Fujii G. Establishment of cultured cell lines derived from a human gastric carcinoma. Jap J Exp Med 1978; 48: 61–68.
Hayashi H, Tatebe S, Osaki M, Goto A, Suzuki Y, Ito H. Expression of Fas antigen and its mediation of apoptosis in human gastric cancer cell lines. Jpn J Cancer Res 1997; 88: 49–55.
Kyo E, Yokozaki H, Yanagihara K, et al. Reduced tumorigenicity and cell motility of a gastric carcinoma cell line by introduction of wild-type p53 gene. Int J Oncol 1993; 3: 265–271.
Tamura T, Aoyama N, Saya H, et al. Induction of Fas-mediated apoptosis in p53-transfected human colon carcinoma cells. Oncogene 1995; 11: 1939–1946.
Polyak K, Waldman T, He TC, Kinzler KW, Vogelstein B. Genetic determinants of p53-induced apoptosis and growth arrest. Genes Dev 1996; 10: 1945–1952.
Brugarolas J, Chandrasekaran C, Gordon JI, Beach D, Jacks T, Hannon GJ. Radiation-induced cell cycle arrest compromised by p21 deficiency. Nature 1995; 377: 552–557.
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Osaki, M., Tatebe, S., Goto, A. et al. 5-Fluorouracil (5-FU) induced apoptosis in gastric cancer cell lines: role of the p53 gene. Apoptosis 2, 221–226 (1997). https://doi.org/10.1023/A:1026476801463
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DOI: https://doi.org/10.1023/A:1026476801463