Abstract
DLC-1 (deleted in liver cancer) gene is frequently deleted in hepatocellular carcinoma. However, little is known about the genetic status and the expression of this gene in gastric cancer. In this study, Northern and Southern analysis showed that seven of nine human gastric cancer cell lines did not express DLC-1 mRNA, but contained the DLC-1 gene. To identify the mechanism of the loss of DLC-1 mRNA expression in these cell lines, we investigated the methylation status of DLC-1 gene by using methylation-specific PCR (MSP) and Southern blot, and found that five of seven DLC-1 nonexpressing gastric cancer cell lines were methylated in the DLC-1 CpG island. Treatment with 5-aza-2′-deoxycytidine (5-Aza-dC) induced DLC-1 mRNA expression in the gastric cancer cell lines that have the methylated alleles. Studies using SNU-601 cell line with methylated DLC-1 alleles revealed that nearly all CpG sites within DLC-1 CpG island were methylated, and that the in vitro methylation of the DLC-1 promoter region is enough to repress DLC-1 mRNA expression, regardless of the presence of transcription factors capable of inducing this gene. In all, 29 of 97 (30%) primary gastric cancers were also shown to be methylated, demonstrating that methylation of the DLC-1 CpG island is not uncommon in gastric cancer. In addition, we demonstrated that DLC-1 mRNA expression was induced, and an increase in the level of acetylated H3 and H4 was detected by the treatment with trichostatin A (TSA) in two DLC-1 nonexpressing cell lines that have the unmethylated alleles. Taken together, the results of our study suggest that the transcriptional silencing of DLC-1, by epigenetic mechanism, may be involved in gastric carcinogenesis.
Similar content being viewed by others
References
Baffa R, Santoro R, Bullrich F, Mandes B, Ishii H and Croce CM . (2000). Clin. Cancer Res., 6, 1372–1377.
Burgers WA, Fuks F and Kouzarides T . (2002). Trends Genet., 18, 275–277.
Cameron EE, Bachman KE, Myohanen S, Herman JG and Baylin SB . (1999). Nat. Genet., 21, 103–107.
Crespo P, Schuebel KE, Ostrom AA, Gutkind JS and Bustelo XR . (1997). Nature, 385, 169–172.
Dobosy JR and Selker EU . (2001). Cell Mol. Life Sci., 58, 721–727.
Doetzlhofer A, Rotheneder H, Lagger G, Koranda M, Kurtev V, Brosch G, Wintersberger E and Seiser C . (1999). Mol. Cell. Biol., 19, 5504–5511.
Emi M, Fujiwara Y, Nakajima T, Tsuchiya E, Tsuda H, Hirohashi S, Maeda Y, Tsuruta K, Miyaki M and Nakamura Y . (1992). Cancer Res., 52, 5368–5372.
Euer N, Schwirzke M, Evtimova V, Burtscher H, Jarsch M, Tarin D and Weidle UH . (2002). Anticancer Res., 22, 733–740.
Fujiwara Y, Ohata H, Emi M, Okui K, Koyama K, Tsuchiya E, Nakajima T, Monden M, Mori T, Kurimasa A, Oshimura M and Nakmura Y . (1994). Genes Chromosomes Cancer, 10, 7–14.
Geiman TM and Robertson KD . (2002). J. Cell Biochem., 87, 117–125.
Guo WH, Weng LQ, Ito K, Chen LF, Nakanishi H, Tatematsu M and Ito Y . (2002). Oncogene, 28, 8351–8355.
Hart MJ, Eva A, Evans T, Aaronson SA and Cerione RA . (1991). Nature, 354, 311–314.
Herman JG, Graff JR, Myohanen S, Nelkin BD and Baylin SB . (1996a). Proc. Natl. Acad. Sci. USA, 93, 9821–9826.
Herman JG, Jen J, Merlo A and Baylin SB . (1996b). Cancer Res., 56, 722–727.
Herman JG, Umar A, Polyak K, Graff JR, Ahuja N, Issa JP, Markowitz S, Willson JK, Hamilton SR, Kinzler KW, Kane MF, Kolodner RD, Vogelstein B, Kunkel TA and Baylin SB . (1998). Proc. Natl. Acad. Sci. USA, 95, 6870–6875.
Huang L, Sowa Y, Sakai T and Pardee AB . (2000). Oncogene, 19, 5712–5719.
Jones PA and Taylor SM . (1980). Cell, 20, 85–93.
Jong HS, Park YI, Kim S, Sohn JH, Kang SH, Song SH, Bang YJ and Kim NK . (1999). Cancer (Philadelphia), 86, 559–565.
Kang SH, Bang YJ, Im YH, Yang HK, Lee DA, Lee HY, Lee HS, Kim NK and Kim S-J . (1999). Oncogene, 18, 7280–7286.
Kikuchi T, Itoh F, Toyota M, Suzuki H, Yamamoto H, Fujita M, Hosokawa M and Imai K . (2002). Int. J. Cancer, 97, 272–277.
Knosel T, Petersen S, Schwabe H, Schluns K, Stein U, Schlag PM, Dietel M and Petersen I . (2002). Virchows Arch., 440, 187–194.
Kurimoto F, Gemma A, Hosoya Y, Seike M, Takenaka K, Uematsu K, Yoshimura A, Shibuya M and Kudoh S . (2001). Int. J. Mol. Med., 8, 89–93.
Morii N, Kumagai N, Nur-E-Kamal MS, Narumiya S and Maruta H . (1993). J. Biol. Chem., 268, 27160–27163.
Nan X, Ng HH, Johnson CA, Laherty CD, Turner BM, Eisenman RN and Bird A . (1998). Nature, 393, 386–389.
Ng IO, Liang ZD, Cao L and Lee TK . (2000). Cancer Res., 60, 6581–6584.
Pannell D, Osborne CS, Yao S, Sukonnik T, Pasceri P, Karaiskakis A, Okano M, Li E, Lipshitz HD and Ellis J . (2000). EMBO J., 19, 5884–5894.
Qiu RG, Chen J, McCormick F and Symons M . (1995). Proc. Natl. Acad. Sci. USA, 92, 11781–11785.
Richon VM, Sandhoff TW, Rifkind RA and Marks PA . (2000). Proc. Natl. Acad. Sci. USA, 97, 10014–10019.
Ridley AJ and Hall A . (1992). Cell, 70, 389–399.
Robertson KD . (2002). Oncogene, 21, 5361–5379.
Satoh A, Toyota M, Itoh F, Kikuchi T, Obata T, Sasaki Y, Suzuki H, Yawata A, Kusano M, Fujita M, Hosokawa M, Yanagihara K, Tokino T and Imai K . (2002). Br. J. Cancer, 86, 1817–1823.
Sekimata M, Kabuyama Y, Emori Y and Homma Y . (1999). J. Biol. Chem., 274, 17757–17762.
Sliva D, Mason R, **ao H and English D . (2000). Biochem. Biophys. Res. Commun., 268, 471–479.
Song SH, Jong HS, Choi HH, Inoue H, Tanabe T, Kim NK and Bang YJ . (2001). Cancer Res., 61, 4628–4635.
Song SH, Jong HS, Choi HH, Kang SH, Ryu MH, Kim NK, Kim WH and Bang YJ . (2000). Int. J. Cancer, 87, 236–240.
Sowa Y, Orita T, Minamikawa S, Nakano K, Mizuno T, Nomura H and Sakai T . (1997). Biochem. Biophys. Res. Commun., 241, 142–150.
Stirzaker C, Millar DS, Paul CL, Warnecke PM, Harrison J, Vincent PC, Frommer M and Clark SJ . (1997). Cancer Res., 57, 2229–2237.
Takakura M, Kyo S, Sowa Y, Wang Z, Yatabe N, Maida Y, Tanaka M and Inoue M . (2001). Nucleic Acids Res., 29, 3006–3011.
Taniura S, Kamitani H, Watanabe T and Eling TE . (2002). J. Biol. Chem., 277, 16823–16830.
Tsutsumi S, Gupta SK, Hogan V, Collard JG and Raz A . (2002). Cancer Res., 62, 4484–4490.
Van Lint C, Emiliani S and Verdin E . (1996). Gene Expression, 5, 245–253.
Vignali M, Hassan AH, Neely KE and Workman JL . (2000). Mol. Cell Biol., 20, 1899–1910.
Wade PA, Jones PL, Vermaak D, Veenstra GJ, Imhof A, Sera T, Tse C, Ge H, Shi YB, Hansen JC and Wolffe AP . (1998). Cold Spring Harb. Symp. Quant. Biol., 63, 435–445.
Wiltshire RN, Dennis TR, Sondak VK and Meltzer PS . (2001). Trent. JM. Cancer Genet. Cytogenet., 131, 97–103.
Wistuba II, Behrens C, Virmani AK, Milchgrub S, Syed S, Lam S, Mackay B, Minna JD and Gazdar AF . (1999). Cancer Res., 59, 1973–1979.
Yaremko ML, Recant WM and Westbrook CA . (1995). Genes Chromosomes Cancer, 13, 186–191.
Yuan BZ, Miller MJ, Keck CL, Zimonjic DB, Thorgeirsson SS and Popescu NC . (1998). Cancer Res., 58, 2196–2199.
Zhang Y and Dufau ML . (2002). J. Biol. Chem., 277, 33431–33438.
Acknowledgements
This work was supported in part by grants from the Ministry of Science & Technology of Korea through the National Research Laboratory Program for Cancer Epigenetics and by 2002 BK21 Project for Medicine, Dentistry, and Pharmacy.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kim, T., Jong, HS., Song, SH. et al. Transcriptional silencing of the DLC-1 tumor suppressor gene by epigenetic mechanism in gastric cancer cells. Oncogene 22, 3943–3951 (2003). https://doi.org/10.1038/sj.onc.1206573
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.onc.1206573
- Springer Nature Limited
Keywords
This article is cited by
-
Impact of clinical features, cytogenetics, genetic mutations, and methylation dynamics of CDKN2B and DLC-1 promoters on treatment response to azacitidine
Annals of Hematology (2020)
-
Epigenetic mechanisms regulating the development of hepatocellular carcinoma and their promise for therapeutics
Hepatology International (2017)
-
Curcumin inhibits growth of human breast cancer cells through demethylation of DLC1 promoter
Molecular and Cellular Biochemistry (2017)
-
Aberrant gene promoter methylation of p16, FHIT, CRBP1, WWOX, and DLC-1 in Epstein–Barr virus-associated gastric carcinomas
Medical Oncology (2015)
-
DLC1 expression is reduced in human cutaneous melanoma and correlates with patient survival
Modern Pathology (2014)