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Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 1996 Aug;87(8):824–830. doi: 10.1111/j.1349-7006.1996.tb02106.x

Genetic Status and Expression of the Cyclin‐dependent Kinase Inhibitors in Human Gastric Carcinoma Cell Lines

Yoshihiko Akama 1,2, Wataru Yasui 2, Hiroki Kuniyasu 2, Hiroshi Yokozaki 2, Morihisa Akagi 2, Hidetoshi Tahara 3, Takenori Ishikawa 1, Eiichi Tahara 2,✉
PMCID: PMC5921170  PMID: 8797888

Abstract

Deregulation of cyclin, cyclin‐dependent kinases (CDKs) and their inhibitors could have a pivotal role in the development of diverse human cancers. We examined the genetic status and the expression of CDK inhibitors (p21, p27, pl6 and p15), CDK2 and cyclins (A, D1 and E) in eight gastric carcinoma cell lines, in comparison with the status of p53 gene alterations. All the cell lines (except MKN‐28) that contained a p53 gene abnormality expressed very low or undetectable levels of p21 mRNA, while the cell lines (MKN‐45 and ‐74) with wild‐type p53 gene expressed high levels of p21 mRNA. An inverse correlation was found between the level of p21 mRNA and the expression of mRNAs for CDK2 and G1 cyclins. MKN‐28 was an exception; it contained mutated p53, and expressed mRNAs for p21, CDK2 and G1 cyclins at high levels. Only MKN‐45 and ‐74, with wild‐type p53, expressed considerable levels of p21 protein. Homozygous deletion of the p16 and p15 genes was detected in two (MKN‐45 and HSC‐39) of the eight gastric carcinoma cell lines. p16 protein was not expressed in three cell lines (MKN‐28, MKN‐74 and KATO‐III), as well as MKN‐45 and HSC‐39. Rearrangement of the p15 gene was found in TMK‐1. Rearrangement of the p27 gene was detected in MKN‐45, although the expression of p27 protein was well preserved in all the gastric carcinoma cell lines. The expression of pRb was also preserved in all the cell lines except KATO‐III. No obvious correlation was observed between the p53 gene status and the expression of p27 and p16. These findings suggest that abnormal regulation of CDK2/cyclins and CDK inhibitors might be involved in deregulated growth of gastric carcinomas.

Keywords: CDK inhibitors, CDK, Cyclin, Cell cycle, Gastric carcinoma cells

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REFERENCE

  • 1.Hunter , T. and Pines , J.Cyclins and cancer II: cyclin D and cdk inhibitors come of age . Cell , 79 , 573 – 582 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 2.Sherr , C. J.Mammalian G1 cyclins . Cell , 73 , 1059 – 1065 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 3.Jiang , W. , Kahn , Ç. M. , Tomita , N. , Zhang , Y.‐J. , Lu , S. H. and Weinstein , I. B.Amplification and expression of the human cyclin D gene in esophageal cancer . Cancer Res ., 52 , 2980 – 2983 ( 1992. ). [PubMed] [Google Scholar]
  • 4.Kitahara , K. , Yasui , W. , Kuniyasu , H. , Yokozaki , H. , Akama , Y. , Yunotani , S. , Hisatsugu , T. and Tahara , E.Concurrent amplification of cyclin E and cdk2 genes in colorectal carcinomas . Int. J. Cancer , 62 , 25 – 28 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 5.Nishida , N. , Fukuda , Y. , Komeda , T. , Kita , R. , Sando , T. , Furukawa , M. , Amenomori , M. , Shibagaki , I. , Nakao , K. , Ikenaga , M. and Ishizaki , K.Amplification and over‐expression of the cyclin D1 gene in aggressive human hepatocellular carcinoma . Cancer Res ., 54 , 3107 – 3110 ( 1994. ). [PubMed] [Google Scholar]
  • 6.Burkley , M. F. , Sweeny , K. J. E. , Hamilton , J. A. , Sini , R. L. , Manning , D. L. , Nicholson , R. I. , deFazio , A. , Watts , C. K. W. , Musgrove , E. A. and Sutherland , R. L.Expression and amplification of cyclin genes in human breast cancer . Oncogene , 8 , 2127 – 2133 ( 1993. ). [PubMed] [Google Scholar]
  • 7.Akama , Y. , Yasui , W. , Yokozaki , H. , Kuniyasu , H. , Kitahara , K. , Ishikawa , T. and Tahara , E.Frequent amplification of the cyclin E gene in human gastric carcinomas . Jpn. J. Cancer Res ., 86 , 617 – 621 ( 1995. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8.Yasui , W. , Ayhan , A. , Kitadai , Y. , Nishimura , K. , Yokozaki , H. , Ito , H. and Tahara , E.Increased expression of p34cdc2 and its kinase activity in human gastric and colonic carcinomas . Int. J. Cancer , 43 , 36 – 41 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 9.Noda , A. , Ning , Y. , Venable , S. F. , Smith , O. M. and Smith , J. R.Cloning of senescent cell derived inhibitors of DNA synthesis using an expression screen . Exp. Cell Res ., 211 , 90 – 98 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 10.Deiry , W. S. , Tokino , T. , Velcuescu , V. E. , Levy , D. B. , Parsons , R. , Trent , J. M. , Lin , D. , Mercer , W. E. , Kinzler , K. W. and Vogelstein , B.WAF1, a potential mediator of p53 tumor suppression . Cell , 75 , 817 – 825 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 11.Harper , J. W. , Adami , G. R. , Wei , N. , Keyomarsi , K. and Elledge , S. J.The p21 cdk‐interacting protein cip 1 is a potent inhibitor of G1 cyclin‐dependent kinase . Cell , 75 , 805 – 816 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 12.Polyak , K. , Lee , M.‐H. , Bromage , H. , Koff , A. , Roberts , J. M. , Tempst , P. and Massague , J.Cloning of p27Kip1, a cyclin‐dependent kinase inhibitor and a potential mediator of extracellular antimitogenic signals . Cell , 78 , 59 – 66 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 13.Toyoshima , H. and Hunter , T.p27, a novel inhibitor of G1 cyclin‐cdk protein kinase activity, is related to p21 . Cell , 78 , 67 – 74 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 14.Kamb , A. , Gruis , N. A. , Feldhaus , J. , Liu , Q. , Harshman , K. , Tavtigian , S. V. , Stockert , E. , Day , R. S. , III , Johnson , B. E. and Skolnick , M. H.A cell cycle regulator potentially involved in genesis of many tumor types . Science , 264 , 436 – 440 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 15.Nobori , T. , Miura , K. , Wu , D. J. , Lois , A. , Takabayashi , K. and Carson , D. A.Deletion of the cyclin‐dependent kinase‐4 inhibitor gene in multiple human cancers . Nature , 368 , 753 – 756 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 16.Waga , S. , Hannon , G. J. , Beach , D. and Stillman , B.The p21 inhibitor of cyclin‐dependent kinases controls DNA replication by interaction with PCNA . Nature , 369 , 574 – 578 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 17.Sun , Y. , Hildesheim , A. , Li , H. , Li , Y. , Chen , J.‐Y. , Chen , Y.‐J. , Hayes , R. B. , Rothman , N. , Bi , W.‐F. , Cao , Y. , Yao , K.‐T. , Lanier , A. P. , Hegamyer , G.E. , Deiry , W. S. , Xiong , Y. and Colburn , N. H.No point mutation but a codon 31ser‐ arg polymorphism of the WAF‐1/CIP1/p21 tumor suppressor gene in nasopharyngeal carcinoma (NPC): the polymorphism distinguishes Caucasians from Chinese . Cancer Epidemiol. Biomarkers Prev ., 4 , 261 – 267 ( 1995. ). [PubMed] [Google Scholar]
  • 18.Koopman , J. , Maintz , D. , Schild , S. , Schramm , J. , Louis , D. N. , Wiestler , O. D. and Deimling , A. V.Multiple polymorphisms, but no mutations, in the WAF1/CIP1 gene in human brain tumours . Br. J. Cancer , 72 , 1230 – 1233 ( 1995. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19.Bhatia , K. , Fan , S. , Spangler , G. , Weintraub , M. , O'Connor , P. M. , Judde , J.‐G. and Magrath , I.A mutant p21 cyclin‐dependent kinase inhibitor isolated from a Burkitt's lymphoma . Cancer Res ., 55 , 1431 – 1435 ( 1995. ). [PubMed] [Google Scholar]
  • 20.Tahara , E.Genetic alterations in human gastrointestinal cancers: the application to molecular diagnosis . Cancer , 75S , 1410 – 1417 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 21.Mattar , R. , Yokozaki , H. , Yasui , W. , Ito , H. and Tahara , E.p53 gene mutations in gastric cancer cell lines . Oncology (Life Sci. Adv.) , 11 , 7 – 12 ( 1992. ). [Google Scholar]
  • 22.Yokozaki , H. , Kuniyasu , H. , Kitadai , Y. , Nishimura , K. , Todo , H. , Ayhan , A. , Yasui , W. , Ito , H. and Tahara , E.p53 point mutations in primary human gastric carcinomas . J. Cancer Res. Clin. Oncol ., 119 , 67 – 70 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 23.Ayhan , A. , Yasui , W. , Yokozaki , H. , Ito , H. and Tahara , E.Genetic abnormalities and expression of p53 in human colon carcinomas . Int. J. Oncol ., 1 , 431 – 437 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 24.Polyak , K. , Kato , J. , Solomon , M. J. , Sherr , C. J. , Massague , J. , Roberts , J. M. and Koff , A.p27Kip1, a cyclin‐cdk inhibitor, links transforming growth factor‐β and contact inhibition to cell cycle arrest . Genes Dev ., 8 , 9 – 22 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 25.Castañeda , M. V. , Lee , M.‐H. , Latres , E. , Polyak , K. , Lacombe , L. , Montgomery , K. , Mathew , S. , Krauter , K. , Sheinfeld , J. , Massague , J. and Cardo , C.P27Kip1: chromosomal mapping to 12p12‐12p13.1 and absence of mutations in human tumors . Cancer Res ., 55 , 1211 – 1214 ( 1995. ). [PubMed] [Google Scholar]
  • 26.Kawamata , N. , Morosetti , R. , Miller , C. W. , Park , D. , Spirin , K. S. , Nakamaki , T. , Takeuchi , S. , Hatta , Y. , Simpson , J. , Wilczynski , S. , Lee , Y. Y. , Bartram , C. R. and Koeffler , H. P.Molecular analysis of the cyclin‐dependent kinase inhibitor gene p27/Kip1 in human malignancies . Cancer Res ., 55 , 2266 – 2269 ( 1995. ). [PubMed] [Google Scholar]
  • 27.Morosetti , R. , Kawamata , N. , Gombart , A. F. , Miller , C. W. , Hatta , Y. , Hirama , T. , Said , J. W. , Tomonaga , M. and Koeffler , H. P.Alterations of the p27Kip1 gene in non‐Hodgkin's lymphomas and adult T‐cell leukemia/lymphoma . Blood , 86 , 1924 – 1930 ( 1995. ). [PubMed] [Google Scholar]
  • 28.Serrano , M. , Hannon , G. J. and Beach , D.A new regulatory motif in cell‐cycle control causing specific inhibition of cyclin D/CDK4 . Nature , 366 , 704 – 707 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 29.Hannon , G. J. and Beach , D.p15INK4D is a potential effector of TGF‐β induced cell cycle arrest . Nature , 371 , 257 – 260 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 30.Mori , T. , Miura , K. , Aoki , T. , Nishihira , T. , Mori , S. and Nakamura , Y.Frequent somatic mutation of the MTS1/CDK4I (multiple tumor suppressor/cyclin‐dependent kinase 4 inhibitor) gene in esophageal squamous cell carcinoma . Cancer Res ., 54 , 3396 – 3397 ( 1994. ). [PubMed] [Google Scholar]
  • 31.Lukas , J. , Parry , D. , Agaard , L. , Mann , D. J. , Bartkova , J. , Straus , M. , Peters , G. and Bartek , J.Retinoblastoma‐protein‐dependent cell‐cycle inhibition by tumour suppressor p16 . Nature , 375 , 503 – 506 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 32.Koh , J. , Enders , G. H. , Dynlacht , B. D. and Harlow , E.Tumour‐derived p16 alleles encoding proteins defective in cell‐cycle inhibition . Nature , 375 , 506 – 510 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 33.Ochiai , A. , Yasui , W. and Tahara , E.Growth‐promoting effect of gastrin on human gastric carcinoma cell line TMK‐1 . Jpn. J. Cancer Res ., 76 , 1064 – 1071 ( 1985. ). [PubMed] [Google Scholar]
  • 34.Yanagihara , K. , Seyama , T. , Tsumuraya , M. , Kamada , N. and Yokoro , K.Establishment and characterization of human signet ring cell gastric carcinoma cell lines with amplification of the c‐myc oncogene . Cancer Res ., 51 , 381 – 386 ( 1991. ). [PubMed] [Google Scholar]
  • 35.Yoshida , K. , Yokozaki , H. , Niimoto , M. , Ito , H. , Ito , M. and Tahara , E.Expression of TGF‐β and procollagen type I and type III in human gastric carcinomas . Int. J. Cancer , 44 , 394 – 398 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 36.Wang , J. , Chenivesse , X. , Henglein , B. and Brechot , C.Hepatitis B virus integration in a cyclin A gene in a hepatocellular carcinoma . Nature , 343 , 555 – 557 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 37.Lew , D. J. , Dulic , V. and Reed , S. I.Isolation of three novel human cyclins by rescue of G1 cyclin (cln) function in yeast . Cell , 66 , 1197 – 1206 ( 1991. ). [DOI] [PubMed] [Google Scholar]
  • 38.Tsai , L.‐H. , Harlow , E. and Meyerson , M.Isolation of the human cdk2 gene that encodes the cyclin A‐ and adenovirus E1A‐associated p33 kinase . Nature , 353 , 174 – 177 ( 1991. ). [DOI] [PubMed] [Google Scholar]
  • 39.Akagi , M. , Yasui , W. , Akama , Y. , Yokozaki , H. , Tahara , H. , Haruma , K. , Kajiyama , G. and Tahara , E.Inhibition of cell growth by transforming growth factor β1 is associated with p53‐independent induction of p21 in gastric carcinoma cells . Jpn. J. Cancer Res ., 87 , 377 – 384 ( 1996. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 40.Bates , S. , Bonetta , L. , MacAllan , D. , Parry , D. , Holder , A. , Dickson , C. and Peters , G.CDK6 (PLSTIRE) and CDK4 (PSK‐J3) are a distinct subset of the cyclin‐dependent kinases that associate with cyclin D1 . Oncogene , 9 , 71 – 79 ( 1994. ). [PubMed] [Google Scholar]
  • 41.Michieli , P. , Chedid , M. , Lin , D. , Pierce , J. H. , Mercer , W. E. and Givol , D.Induction of Waf1/CIP1 by a p53‐independent pathway . Cancer Res ., 54 , 3391 – 3395 ( 1994. ). [PubMed] [Google Scholar]
  • 42.Elbendary , A. , Berchuck , A. , Davis , P. , Havrilesky , L. , Bast , R. C. and Igrehart , J. D.Transforming growth factor β1 can induce CIP1/WAF1 expression independent of the p53 pathway in ovarian cancer cells . Cell Growth Differ ., 5 , 1301 – 1307 ( 1994. ). [PubMed] [Google Scholar]
  • 43.Deng , C. , Zhang , P. , Harper , J. W. , Elledge , S. J. and Leder , P.Mice lacking p21CIP1/WAF1 undergo normal development, but are defective in G1 checkpoint control . Cell , 82 , 675 – 684 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 44.Spruck , C. H. I. , Zulueta , M. , Shibata , A. , Simoneau , A. R. , Lin , M.‐F. , Gonzales , F. , Tsai , Y. C. and Jones , P. A.p16 gene in uncultured tumours . Nature , 370 , 183 – 184 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 45.Zhang , S. Y. , Szanto , A. J. , Sauter , E. R. , Shafarenko , M. , Mitsunaga , S. , Nobori , T. , Carson , D. A. , Ridge , J. A. and Goodrow , T. L.Higher frequency of alterations in the p16/CDKN2 gene in squamous cell carcinoma cell lines than in primary tumors of the head and neck . Cancer Res ., 54 , 5050 – 5053 ( 1994. ). [PubMed] [Google Scholar]
  • 46.Merlo , A. , Herman , J. G. , Lee , D. J. , Garbrielson , E. , Burger , P. C. , Baylin , S. B. and Sidransky , D.5′ CpG island methylation is associated with transcriptional silencing of the tumour suppressor p16/CDKN2/MTS1 in human cancers . Nat. Med ., 1 , 686 – 692 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 47.Zulueta , M. , Bender , C. M. , Yang , A. S. , Nguyen , T. , Beart , R. W. , Tornout , J. M. V. and Jones , P. A.Methylation of the 5′ CpG island of the p16/CDKN2 tumor suppressor gene in normal and transformed human tissues with gene silencing . Cancer Res ., 55 , 4531 – 4535 ( 1995. ). [PubMed] [Google Scholar]
  • 48.Otterson , G. A. , Kratzke , R. A. , Coxon , A. , Kim , Y. W. and Kaye , F. J.Absence of p16INK4 protein is restricted to the subset of lung cancer lines that retains wild type Rb . Oncogene , 9 , 3375 – 3378 ( 1994. ). [PubMed] [Google Scholar]
  • 49.Yeager , T. , Stadler , W. , Belair , C. , Puthenveettil , J. , Olopade , O. and Reznikoff , C.Increased p16 levels correlate with pRb alterations in human urothelial cells . Cancer Res ., 55 , 493 – 497 ( 1995. ). [PubMed] [Google Scholar]
  • 50.Weinberg , R. A.The retinoblastoma protein and cell cycle control . Cell , 81 , 323 – 330 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 51.Kitahara , K. , Yasui , W. , Yokozaki , H. , Semba , S. , Hamamoto , T. , Hisatsugu , T. and Tahara , E.Expression of cyclin D1, CDK4 and p27KIP1 is associated with the p16MTS1 gene status in human esophageal carcinoma cell lines . J. Exp. Ther. Oncol ., 1 , 7 – 12 ( 1996. ). [PubMed] [Google Scholar]

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