Skip to main content

Advertisement

Log in

The impact of C-MYC gene expression on gastric cancer cell

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

The upregulation or mutation of C-MYC has been observed in gastric, colon, breast, and lung tumors and in Burkitt’s lymphoma. However, little is known about the role C-MYC plays in gastric adenocarcinoma. In the present study, we intended to investigate the influence of C-MYC on the growth, proliferation, apoptosis, invasion, and cell cycle of the gastric cancer cell line SGC7901 and the gastric cell line HFE145. C-MYC cDNA was subcloned into a constitutive vector PCDNA3.1 followed by transfection in normal gastric cell line HFE145 by using liposome. Then stable transfectants were selected and appraised. Specific inhibition of C-MYC was achieved using a vector-based siRNA system which was transfected in gastric cancer cell line SGC7901. The apoptosis and cell cycles of these clones were analyzed by using flow cytometric assay. The growth and proliferation were analyzed by cell growth curves and colony-forming assay, respectively. The invasion of these clones was analyzed by using cell migration assay. The C-MYC stable expression clones (HFE-Myc) and C-MYC RNAi cells (SGC-MR) were detected and compared with their control groups, respectively. HFE-Myc grew faster than HFE145 and HFE-PC (HFE145 transfected with PCDNA3.1 vector). SGC-MR1, 2 grew slower than SGC7901 and SGC-MS1, 2 (SGC7901 transfected with scrambled control duplexes). The cell counts of HFE-Myc in the third, fourth, fifth, sixth, and seventh days were significantly more than those of control groups (P < 0.05). Those of SGC-MR1, 2 in the fourth, fifth, sixth, and seventh days were significantly fewer than those of control groups (P < 0.05). Cell cycle analysis showed that proportions of HFE-Myc and SGC-MR cells in G0–G1 and G2–M were different significantly with their control groups, respectively (P < 0.05). The apoptosis rate of HFE-Myc was significantly higher than those of control groups (P < 0.05). Results of colony-forming assay showed that the colony formation rate of HFE-Myc was higher than those of control groups; otherwise, the rate of SGC-MR was lower than those of their control groups (P < 0.05). The results of cell migration assay showed that there were no significant differences between experimental groups and control groups (P > 0.05). In conclusion, C-MYC can promote the growth and proliferation of normal gastric cells, and knockdown of C-MYC can restrain the growth and proliferation of gastric cancer cells. It can induce cell apoptosis and help tumor cell maintain malignant phenotype. But it can have not a detectable influence on the ability of invasion of gastric cancer cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10

Similar content being viewed by others

References

  1. Parkin DM, Poisani P, Ferlay (1999) Global cancer statistics. CA Cancer J Clin 49(1):33–64

    Article  CAS  PubMed  Google Scholar 

  2. Correa P (1992) Human gastric carcinogenesis: a multistep and multifactorial process—First American Cancer society award lecture on cancer epidemiology and prevention. Cancer Res 52(24):6735–6740

    CAS  PubMed  Google Scholar 

  3. Gonzalez CA, Sala N, Capella G (2002) Genetic susceptibility and gastric cancer risk. Int J Cancer 100(3):249–260

    Article  CAS  PubMed  Google Scholar 

  4. Xue FB, Xu YY, Wan Y et al (2001) Association of H. pylori infection with gastric carcinoma: a meta analysis. World J Gastroenterol 7(6):801–804

    CAS  PubMed  Google Scholar 

  5. Nesbit CE, Tersak JM, Prochownik EV (1999) MYC oncogenes and human neoplastic disease. Oncogene 18(19):3004–3016

    Article  CAS  PubMed  Google Scholar 

  6. Dang CV (1999) C-MYC target genes involved in cell growth, apoptosis, an metabolism. Mol Cell Biol 19(1):1–11

    CAS  PubMed  Google Scholar 

  7. Cole MD, McMahon SB (1999) The Myc oncoprotein: a critical evaluation of transactivation and target gene regulation. Oncogene 18(19):2916–2924

    Article  CAS  PubMed  Google Scholar 

  8. Obaya AJ, Mateyak MK, Sedivy JM (1999) Mysterious liaisons: the relationship between C-MYC and the cell cycle. Oncogene 18(19):2934–2941

    Article  CAS  PubMed  Google Scholar 

  9. Hoffman B, Liebermann DA (1998) The proto-oncogene c-myc and apoptosis. Oncogene 17(25):3351–3357

    Article  PubMed  Google Scholar 

  10. Prendergast GC (1999) Mechanisms of apoptosis by C-Myc. Oncogene 18(19):2967–2987

    Article  CAS  PubMed  Google Scholar 

  11. Dang CV, Resar LM, Emison E et al (1999) Function of the C-MYC oncogenic transcription factor. Exp Cell Res 253(1):63–77

    Article  CAS  PubMed  Google Scholar 

  12. Milne AN, Sitarz R, Carvalho R et al (2007) Early onset gastric cancer: on the road to unraveling gastric carcinogenesis. Curr Mol Med 7(1):15–28

    Article  CAS  PubMed  Google Scholar 

  13. Calcagno DQ, Leal MF, Seabra AD et al (2006) Interrelationship between chromosome 8 aneuploidy, C-MYC amplification and increased expression in individuals from northern Brazil with gastric adenocarcinoma. World J Gastroenterol 12(38):6207–6211

    CAS  PubMed  Google Scholar 

  14. Kozma L, Kiss I, Hajdu J et al (2001) C-myc amplification and cluster analysis in human gastric carcinoma. Anticancer Res 21(1B):707–710

    CAS  PubMed  Google Scholar 

  15. Yang GF, Deng CS, Xiong YY et al (2004) Expression of nuclear factor-kappa B and target genes in gastric precancerous lesions and adenocarcinoma: association with Helicobactor pylori cagA (+) infection. World J Gastroenterol 10(4):491–496

    CAS  PubMed  Google Scholar 

  16. Onoda N, Maeda K, Chung YS et al (1996) Overexpression of c-myc messenger RNA in primary and metastatic lesions of carcinoma of the stomach. J Am Coll Surg 182(1):55–59

    CAS  PubMed  Google Scholar 

  17. Han S, Kim HY, Park K et al (1999) c-Myc expression is related with cell proliferation and associated with poor clinical outcome in human gastric cancer. J Korean Med Sci 14(5):526–530

    CAS  PubMed  Google Scholar 

  18. Nakata B, Onoda N, Chung YS et al (1995) Correlation between malignancy of gastric cancer and c-myc DNA amplification or overexpression of c-myc protein. Gan To Kagaku Ryoho 22(Suppl 2):176–179

    CAS  PubMed  Google Scholar 

  19. Sanz-Ortega J, Steinberg SM, Moro E et al (2000) Comparative study of tumor angiogenesis and immunohistochemistry for p53, c-ErbB2, c-myc and EGFr as prognostic factors in gastric cancer. Histol Histopathol 15(2):455–462

    CAS  PubMed  Google Scholar 

  20. Ishii HH, Gobe GC, Pan W et al (2002) Apoptosis and cell proliferation in the development of gastric carcinomas: associations with c-myc and p53 protein expression. J Gastroenterol Hepatol 17(9):966–972

    Article  CAS  PubMed  Google Scholar 

  21. Costa Raiol LC, Figueira Silva EC, Mendes da Fonseca D et al (2008) Interrelationship between MYC gene numerical aberrations and protein expression in individuals from northern Brazil with early gastric adenocarcinoma. Cancer Genet Cytogenet 181(1):31–35

    Article  PubMed  Google Scholar 

  22. Xu AG, Li SG, Liu JH et al (2001) Function of apoptosis and expression of the proteins Bcl-2, p53 and C-myc in the development of gastric cancer. World J Gastroenterol 7(3):403–406

    CAS  PubMed  Google Scholar 

  23. Lan J, Xiong YY, Lin YX et al (2003) Helicobacter pylori infection generated gastric cancer through p53-Rb tumor-suppressor system mutation and telomerase reactivation. World J Gastroenterol 9(1):54–58

    CAS  PubMed  Google Scholar 

  24. Hashimoto K, Nakagawa Y, Morikawa H et al (2001) Co-overexpression of DEAD box protein rck/p54 and c-myc protein in human colorectal adenomas and the relevance of their expression in cultured cell lines. Carcinogenesis 22(12):1965–1970

    Article  CAS  PubMed  Google Scholar 

  25. Henriksson M, Luscher B (1996) Proteins of the Myc network: essential regulators of cell growth and differentiation. Adv Cancer Res 68:109–182

    Article  CAS  PubMed  Google Scholar 

  26. Oster SK, Ho CS, Soucie EL et al (2002) The myc oncogene: MarvelouslY Complex. Adv Cancer Res 84:81–154

    Article  CAS  PubMed  Google Scholar 

  27. Evan GI, Wyllie AH, Gilbert CS et al (1992) Induction of apoptosis in fibroblasts by c-myc protein. Cell 69(1):119–128

    Article  CAS  PubMed  Google Scholar 

  28. Langlois NE, Lamb J, Eremin O et al (1997) Apoptosis in colorectal carcinoma occurring in patients aged 45 years and under: relationship to prognosis, mitosis, and immunohistochemical demonstration of p53, c-myc and bcl-2 protein products. J Pathol 182(4):392–397

    Article  CAS  PubMed  Google Scholar 

  29. Xiangming C, Natsugoe S, Takao S et al (2001) Preserved Smad4 expression in the transforming growth factor beta signaling pathway is a favorable prognostic factor in patients with advanced gastric cancer. Clin Cancer Res 7(2):277–282

    CAS  PubMed  Google Scholar 

  30. Vaux DL, Cory S, Adams JM (1988) Bcl-2 gene promotes haemopoietic cell survival and cooperates with c-myc to immortalize pre-B cells. Nature 335(6189):440–442

    Article  CAS  PubMed  Google Scholar 

  31. Fernandez PC, Frank SR, Wang L et al (2003) Genomic targets of the human c-Myc protein. Genes Dev 17(9):1115–1129

    Article  CAS  PubMed  Google Scholar 

  32. Robinson K, Asawachaicharn N, Denise A (2009) c-Myc accelerates S-phase and requires WRN to avoid replication stress. PLoS One 4(6):e5951

    Article  PubMed  Google Scholar 

  33. Bouchard C, Dittrich O, Kiermaier A et al (2001) Regulation of cyclin D2 gene expression by the Myc/Max/Mad network: Myc-dependent TRRAP recruitment and histone acetylation at the cyclin D2 promoter. Genes Dev 15(16):2042–2047

    Article  CAS  PubMed  Google Scholar 

  34. Menssen A, Hermeking H (2002) Characterization of the c-MYC-regulated transcriptome by SAGE: identification and analysis of c-MYC target genes. Proc Natl Acad Sci USA 99(9):6274–6279

    Article  CAS  PubMed  Google Scholar 

  35. Hermeking H, Rago C, Schuhmacher M et al (2000) Identification of CDK4 as a target of c-MYC. Proc Natl Acad Sci USA 97(5):2229–2234

    Article  CAS  PubMed  Google Scholar 

  36. Orian A, van Steensel B, Delrow J et al (2003) Genomic binding by the Drosophila Myc, Max, Mad/Mnt transcription factor network. Genes Dev 17(9):1101–1114

    Article  CAS  PubMed  Google Scholar 

  37. Miliani de Marval PL, Macias E, Rounbehler R et al (2004) Lack of cyclin-dependent kinase 4 inhibits c-myc tumorigenic activities in epithelial tissues. Mol Cell Biol 24(17):7538–7547

    Article  PubMed  Google Scholar 

  38. Kozar K, Ciemerych MA, Rebel VI et al (2004) Mouse development and cell proliferation in the absence of d-cyclins. Cell 118(4):477–491

    Article  CAS  PubMed  Google Scholar 

  39. Wu S, Cetinkaya C, Munoz-Alonso MJ, von der Lehr N et al (2003) Myc represses differentiation -induced p21CIP1 expression via Miz-1-dependent interaction with the p21 core promoter. Oncogene 22(3):351–360

    Article  CAS  PubMed  Google Scholar 

  40. Seoane J, Pouponnot C, Staller P et al (2001) TGFbeta influences Myc, Miz-1 and Smad to control the CDK inhibitor p15INK4b. Nat Cell Biol 3(4):400–408

    Article  CAS  PubMed  Google Scholar 

  41. Claassen GF, Hann SR (2000) A role for transcriptional repression of p21CIP1 by c-Myc in overcoming transforming growth factor beta induced cell-cycle arrest. Proc Natl Acad Sci USA 97(17):9498–9503

    Article  CAS  PubMed  Google Scholar 

  42. Peltenburg LT, de Bruin EC, Meersma D et al (2004) c-Myc is able to sensitize human melanoma cells to diverse apoptotic triggers. Melanoma Res 14(1):3–12

    Article  CAS  PubMed  Google Scholar 

  43. Pelengaris S, Khan M, Evan G (2002) c-MYC: more than just a matter of life and death. Nat Rev Cancer 2(10):764–776

    Article  CAS  PubMed  Google Scholar 

  44. Nilsson JA, Cleveland JL (2003) Myc pathways provoking cell suicide and cancer. Oncogene 22(56):9007–9021

    Article  CAS  PubMed  Google Scholar 

  45. Yang BS, Geddes TJ, Pogulis RJ et al (1991) Transcriptional suppression of cellular gene expression by c-Myc. Mol Cell Biol 11(4):2291–2295

    CAS  PubMed  Google Scholar 

  46. Yang BS, Gilbert JD, Freytag SO et al (1993) Overexpression of Myc suppresses CCAAT transcription factor/nuclear factor 1-dependent promoters in vivo. Mol Cell Biol 13(5):3093–3102

    CAS  PubMed  Google Scholar 

  47. Shiio Y, Donohoe S, Yi EC et al (2002) Quantitative proteomic analysis of Myc oncoprotein function. EMBO J 21(19):5088–5096

    Article  CAS  PubMed  Google Scholar 

  48. Geisler JP, Geisler HE, Manahan KJ et al (2004) Nuclear and cytoplasmic c-myc staining in endometrial carcinoma and their relationship to survival. Int J Gynecol Cancer 14(1):133–137

    Article  CAS  PubMed  Google Scholar 

  49. Ruzinova MB, Caron T, Rodig SJ (2010) Altered Subcellular Localization of c-Myc Protein Identifies Aggressive B-cell Lymphomas Harboring a c-MYC Translocation. Am J Surg Pathol 34(6):882–891

    Google Scholar 

Download references

Acknowledgments

The authors wish to thank Drs. Haili Huang and Gangshi Wang, and Nurse Weidi You, Weihua Wang et al., for handling patient contacts. We wish to thank the Forth Military Medical University of PLA for providing means for the current investigation.

Conflict of interest statement

The authors declare no competing interest.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Lin Zhang.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhang, L., Hou, Y., Ashktorab, H. et al. The impact of C-MYC gene expression on gastric cancer cell. Mol Cell Biochem 344, 125–135 (2010). https://doi.org/10.1007/s11010-010-0536-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-010-0536-0

Keywords

Navigation