p53 functional activation is independent of its
genotype in five esophageal squamous cell carcinoma cell lines
p53 functional activation is independent of its
genotype in five esophageal squamous cell carcinoma cell lines
Junfang JI,Kun WU,Min WU,Qimin ZHAN,
State Key Laboratory
of Molecular Oncology, Cancer Institute, Chinese Academy of Medical
Sciences & Peking Union Medical College, Beijing 100021, China;
Abstract:p53 mutations have been found in many esophageal squamous cell carcinoma (ESCC) clinical specimens and cell lines. We reasoned that functional inactivation of wild-type p53 or the functional activation of mutant-type p53 might exist in these specimens and cell lines. In this study, we identified the correlation between p53 functional activation and its genotype in five different ESCC cell lines. To examine the potential p53 activation in a certain ESCC cell line, DNA damage methods including X-ray exposure and cisplatin treatment were employed to treat cells. Further, the expression of p53 protein and four transcripts of well-known p53 target genes were investigated using Western blot and reverse transcription-polymerase chain reaction (RT-PCR) after cell exposure to DNA damage. The results showed that in KYSE 30 cell line with mutant p53 and KYSE 150 with wild-type p53, p53 could be activated by DNA damages. However, p53 could not be activated following the DNA damages in YES 2 with wild-type p53, KYSE 70 with mutant p53, and EC9706 with unknown p53 genotype. All our data indicated that p53 function in certain cells is not closely correlated with its genotype. To judge p53 function in a particular cell line, it is important to examine the p53 functional activation, but not to simply rely on the p53 genotype.
. p53 functional activation is independent of its
genotype in five esophageal squamous cell carcinoma cell lines[J]. Front. Med., 2010, 4(4): 412-418.
Junfang JI, Kun WU, Min WU, Qimin ZHAN, . p53 functional activation is independent of its
genotype in five esophageal squamous cell carcinoma cell lines. Front. Med., 2010, 4(4): 412-418.
Parkin D M, Bray F, Ferlay J, Pisani P. Global cancer statistics, 2002. CA Cancer J Clin, 2005, 55(2): 74–108 PMID: 15761078
Ke L. Mortality and incidence trends from esophagus cancerin selected geographic areas of China circa 1970-90. Int J Cancer, 2002, 102(3): 271–274 PMID: 12397650
Lerut T, Coosemans W, De Leyn P, Van Raemdonck D, Deneffe G, Decker G. Treatment of esophageal carcinoma. Chest, 1999, 116(6 Suppl): 463S–465S PMID: 10619509
Zhan Q, Carrier F, Fornace A J Jr. Induction of cellularp53 activity by DNA-damaging agents and growth arrest. Mol Cell Biol, 1993, 13(7): 4242–4250 PMID: 8321226
Hainaut P, Hernandez T, Robinson A, Rodriguez-Tome P, Flores T, Hollstein M, Harris C C, Montesano R. IARC Database of p53 gene mutations in human tumors and cell lines:updated compilation, revised formats and newvisualisation tools. Nucleic Acids Res, 1998, 26(1): 205–213 PMID: 9399837
May P, May E. Twenty years of p53 research: structural and functional aspects of the p53 protein. Oncogene, 1999, 18(53): 7621–7636 PMID: 10618702
Zhan Q, Fan S, Bae I, Guillouf C, Liebermann D A, O'Connor P M, Fornace A J Jr. Induction of bax by genotoxic stress in human cells correlates withnormal p53 status and apoptosis. Oncogene, 1994, 9(12): 3743–3751 PMID: 7970735
Greenblatt M S, Bennett W P, Hollstein M, Harris C C. Mutations in the p53 tumor suppressor gene: clues tocancer etiology and molecular pathogenesis. Cancer Res, 1994, 54(18): 4855–4878 PMID: 8069852
Tanaka H, Shibagaki I, Shimada Y, Wagata T, Imamura M, Ishizaki K. Characterization of p53 genemutations in esophageal squamous cell carcinoma cell lines: increasedfrequency and different spectrum of mutations from primary tumors. Int J Cancer, 1996, 65(3): 372–376 PMID: 8575860
Ko L J, Prives C. p53: puzzle and paradigm. Genes Dev, 1996, 10(9): 1054–1072 PMID: 8654922
Merchant A K, Loney T L, Maybaum J. Expression of wild-type p53 stimulates an increase in both Bax and Bcl-xL protein content in HT29 cells. Oncogene, 1996, 13(12): 2631–2637 PMID: 9000137
Miyashita T, Krajewski S, Krajewska M, Wang H G, Lin H K, Liebermann D A, Hoffman B, Reed J C. Tumor suppressorp53 is a regulator of bcl-2 and bax gene expression in vitro and in vivo. Oncogene, 1994, 9(6): 1799–1805 PMID: 8183579
Pietenpol J A, Tokino T, Thiagalingam S, el-Deiry W S, Kinzler K W, Vogelstein B. Sequence-specific transcriptionalactivation is essential for growth suppression by p53. Proc Natl Acad Sci U S A, 1994, 91(6): 1998–2002 PMID: 8134338
Moll U M, LaQuaglia M, Bénard J, Riou G. Wild-type p53 protein undergoes cytoplasmic sequestrationin undifferentiated neuroblastomas but not in differentiated tumors. Proc Natl Acad Sci U S A, 1995, 92(10): 4407–4411 PMID: 7753819
Moll U M, Ostermeyer A G, Haladay R, Winkfield B, Frazier M, Zambetti G. Cytoplasmic sequestrationof wild-type p53 protein impairs the G1 checkpoint after DNA damage. Mol Cell Biol, 1996, 16(3): 1126–1137 PMID: 8622657
Fan W, Jin S, Tong T, Zhao H, Fan F, Antinore M J, Rajasekaran B, Wu M, Zhan Q. BRCA1 regulates GADD45 through its interactions withthe OCT-1 and CAAT motifs. J Biol Chem, 2002, 277(10): 8061–8067 PMID: 11777930
Barnas C, Martel-Planche G, Furukawa Y, Hollstein M, Montesano R, Hainaut P. Inactivation of the p53 proteinin cell lines derived from human esophageal cancers. Int J Cancer, 1997, 71(1): 79–87 PMID: 9096669
Fujii T, Kato S, Yamana H, Tanaka Y, Fujita H, Shirouzu K, Morimatsu M. Expression of G1 cell cycle markers and the effect of adenovirus-mediatedoverexpression of p21Waf-1 in squamous cell carcinoma of the esophagus. Int J Oncol, 2001, 18(1): 157–163 PMID: 11115554
Rigberg D A, Centeno J, Kim F S, Ke B, Swenson K, Maggard M, McFadden D W. Irradiation-induced up-regulation of Fas in esophagealsquamous cell carcinoma is not accompanied by Fas ligand-mediatedapoptosis. J Surg Oncol, 1999, 71(2): 91–96 PMID: 10389864
el-Deiry W S. Regulation of p53 downstream genes. Semin Cancer Biol, 1998, 8(5): 345–357 PMID: 10101800
Leng R P, Lin Y, Ma W, Wu H, Lemmers B, Chung S, Parant J M, Lozano G, Hakem R, Benchimol S. Pirh2, a p53-induced ubiquitin-proteinligase, promotes p53 degradation. Cell, 2003, 112(6): 779–791 PMID: 12654245
Zhan Q. Gadd45a, a p53- and BRCA1-regulated stress protein, incellular response to DNA damage. Mutat Res, 2005, 569(1–2): 133–143 PMID: 15603758
Rigberg D A, Kim F S, Blinman T A, Cole M A, Lane J S, So J, McFadden D W. p21 expression is increased by irradiation in esophageal squamouscell carcinoma. J Surg Res, 1998, 76(2): 137–142 PMID: 9698513
el-Deiry W S, Kern S E, Pietenpol J A, Kinzler K W, Vogelstein B. Definition of a consensus binding site for p53. Nat Genet, 1992, 1(1): 45–49 PMID: 1301998
Hollstein M, Sidransky D, Vogelstein B, Harris C C. p53 mutations in human cancers. Science, 1991, 253(5015): 49–53 PMID: 1905840
Liang S H, Clarke M F. Regulation of p53 localization. Eur J Biochem, 2001, 268(10): 2779–2783 PMID: 11358492