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Journal of Genetics

, Volume 97, Issue 5, pp 1155–1168 | Cite as

Roles of Cyclin A, Myc, Jun and Ppm1l in tumourigenic transformation of NIH3T3 cell

  • Cuifang Chang
  • Lingling Xi
  • Jihong ZhangEmail author
  • Weiming Zhao
  • Zhiyou Liu
  • Jianlin Guo
  • Cunshuan Xu
Research Article

Abstract

To analyse the mechanism of tumourigenic transformation of NIH3T3 cells at the transcriptional level, we used cancerogen 3-methylcholanthrene (3-MCA) and cancerogenic substance phorbol-12-myristate-13-acetate (PMA) to transform NIH3T3 cells and the assessment of transformation was performed using Giemsa staining and methylcellulose colony formation assay. Changes in gene expression profile were detected by Mouse Genome 430 2.0 microarray; and quantitative real-time polymerase chain reaction and Western blotting were used to verify the expression changes of mRNAs and proteins, respectively. With the aid of bioinformatics method, five signalling pathways were identified to participate in different stages of NIH3T3 cell transformation. Further, our study suggested that oncogenes Cyclin A, Myc, Jun and the tumour suppressor gene Ppm1l may play important roles in these pathways.

Keywords

tumourigenic transformation oncogenes tumour suppressor genes signalling transduction gene expression profile 

Notes

Acknowledgements

This work was supported by the Natural Science Foundation of China (no. 31572270), the Natural Science Foundation of Henan (no. 162300410144) and the Natural Science Foundation of Henan (no. 182300410335).

Supplementary material

12041_2018_1009_MOESM1_ESM.pdf (30 kb)
Supplementary material 1 (pdf 29 KB)

References

  1. Adjei A. A. 2001 Blocking oncogenic Ras signaling for cancer therapy. J. Natl. Cancer Inst. 93, 1062–1074.CrossRefGoogle Scholar
  2. Amato K. R., Wang S., Hastings A. K., Youngblood V. M., Santapuram P. R., Chen H. et al. 2014 Genetic and pharmacologic inhibition of EPHA2 promotes apoptosis in NSCLC. J. Clin. Investig. 124, 2037–2049.CrossRefGoogle Scholar
  3. Antonov A. V., Schmidt E. E., Dietmann S., Krestyaninova M. and Hermjakob H. 2010 R spider: a network-based analysis of gene lists by combining signaling and metabolic pathways from Reactome and KEGG databases. Nucleic Acids Res. 38, W78–W83.CrossRefGoogle Scholar
  4. Bottje W. G., Kong B. W., Song J. J., Lee J. Y., Hargis B. M., Lassiter K. et al. 2012 Gene expression in breast muscle associated with feed efficiency in a single male broiler line using a chicken 44K microarray. II. Differentially expressed focus genes. Poult. Sci. 91, 2576–2587.CrossRefGoogle Scholar
  5. Chang W. Y., Bryce D. M., D’Souza S. J. and Dagnino L. 2004 The DP-1 transcription factor is required for keratinocyte growth and epidermal stratification. J. Biol. Chem. 279, 51343–51353.CrossRefGoogle Scholar
  6. Chen G. Qiu H., Ke S., Hu S., Yu S., Zou S. et al. 2013a The fibroblast growth factor receptor 2-mediated extracellular signal-regulated kinase 1/2 signaling pathway plays is important in regulating excision repair cross-complementary gene 1 expression in hepatocellular carcinoma. Biomed. Rep. 1, 604–608.CrossRefGoogle Scholar
  7. Chen J. S., Hung W. S., Chan H. H., Tsai S. J. and Sun H. S. 2013b In silico identification of oncogenic potential of fyn-related kinase in hepatocellular carcinoma. Bioinformatics 29, 420–427.CrossRefGoogle Scholar
  8. Chen L., Hayden M. S., Gilmore E. S., Alexander-Savino C., Oleksyn D., Gillespie K. et al. 2017 PKK deletion in basal keratinocytes promotes tumorigenesis after chemical carcinogenesis. Carcinogenesis 8, 418–428.Google Scholar
  9. Cheng F., McLaughlin P. J., Verderame M. F. and Zagon I. S. 2008 The OGF-OGFr axis utilizes the p21 pathway to restrict progression of human pancreatic cancer. Mol. Cancer 7, 5.CrossRefGoogle Scholar
  10. Connelly L., Barham W., Onishko H. M., Sherrill T., Chodosh L. A., Blackwell T. S. et al. 2011 Inhibition of NF-kappa B activity in mammary epithelium increases tumor latency and decreases tumor burden. Oncogene 30, 1402–1412.CrossRefGoogle Scholar
  11. Creton S., Aardema M. J., Carmichael P. L., Harvey J. S., Martin F. L., Newbold R. F. et al. 2012 Cell transformation assays for prediction of carcinogenic potential: state of the science and future research needs. Mutagenesis 27, 93–101.CrossRefGoogle Scholar
  12. Donahue A. C. and Fruman D. A. 2004 PI3K signaling controls cell fate at many points in B lymphocyte development and activation. Semin. Cell Dev. Biol. 15, 183–197.CrossRefGoogle Scholar
  13. Ehrhardt C. and Ludwig S. 2009 A new player in a deadly game: influenza viruses and the PI3K/Akt signalling pathway. Cell. Microbiol. 11, 863–871.CrossRefGoogle Scholar
  14. Fong C. W., Chua M. S., McKie A. B., Ling S. H., Mason V., Li R. et al. 2006 Sprouty 2, an inhibitor of mitogen-activated protein kinase signaling, is down-regulated in hepatocellular carcinoma. Cancer Res. 66, 2048–2058.CrossRefGoogle Scholar
  15. Gao Y. and Lui W. Y. 2014 Synergistic effect of interferon-gamma and tumor necrosis factor-alpha on coxsackievirus and adenovirus receptor expression: an explanation of cell sloughing during testicular inflammation in mice. Biol. Reprod. 90, 59.CrossRefGoogle Scholar
  16. Gilan O., Diesch J., Amalia M., Jastrzebski K., Chueh A. C., Verrills N. M. D. et al. 2015 PR55alpha-containing protein phosphatase 2A complexes promote cancer cell migration and invasion through regulation of AP-1 transcriptional activity. Oncogene 34, 1333–1339.CrossRefGoogle Scholar
  17. Gushiken F. C., Patel V., Liu Y., Pradhan S., Bergeron A. L., Peng Y. et al. 2008 Protein phosphatase 2A negatively regulates integrin alpha(IIb)beta(3) signaling. J. Biol. Chem. 283, 12862–12869.CrossRefGoogle Scholar
  18. Hanahan D. and Weinberg R. A. 2000 The hallmarks of cancer. Cell 100, 57–70.CrossRefGoogle Scholar
  19. He X., Tan X., Wang X., Jin H., Liu L., Ma L. et al. 2014 C-Myc-activated long noncoding RNA CCAT1 promotes colon cancer cell proliferation and invasion. Tumour Biol. 35, 12181–12188.CrossRefGoogle Scholar
  20. Khan S., Lopez-Dee Z., Kumar R. and Ling J. 2013 Activation of NFkB is a novel mechanism of pro-survival activity of glucocorticoids in breast cancer cells. Cancer Lett. 337, 90–95.CrossRefGoogle Scholar
  21. Kim S. G., Lee A. J., Bae S. H., Kim S. M., Lee J. H., Kim M. J. et al. 2014 Total extract of Korean red ginseng facilitates human bone marrow hematopoietic colony formation in vitro. Blood Res. 49, 177–181.CrossRefGoogle Scholar
  22. Koso H., Tsuhako A., Lyons E., Ward J. M., Rust A. G., Adams D. J. et al. 2014 Identification of FoxR2 as an oncogene in medulloblastoma. Cancer Res. 74, 2351–2361.CrossRefGoogle Scholar
  23. Lai K. O., Chen Y., Po H. M., Lok K. C., Gong K., Ip N. Y. et al. 2004 Identification of the Jak/Stat proteins as novel downstream targets of EphA4 signaling in muscle: implications in the regulation of acetylcholinesterase expression. J. Biol. Chem. 279, 13383–13392.CrossRefGoogle Scholar
  24. Mori S., Bernardi R., Laurent A., Resnati M., Crippa A., Gabrieli A. et al. 2012 Myb-binding protein 1A (MYBBP1A) is essential for early embryonic development, controls cell cycle and mitosis, and acts as a tumor suppressor. PLoS one 7, e39723.CrossRefGoogle Scholar
  25. Nie J., Liu S. and Di L. 2000 Cancer pain and its influence on cancer patients’ quality of life. Zhonghua Zhong Liu Za Zhi 22, 432–434.PubMedGoogle Scholar
  26. Poburski D. and Thierbach R. 2016 Improvement of the BALB/c-3T3 cell transformation assay: a tool for investigating cancer mechanisms and therapies. Sci. Rep. 6, 32966.CrossRefGoogle Scholar
  27. Quelo I., Gauthier C., Hannigan G. E., Dedhar S. and St-Arnaud R. 2004 Integrin-linked kinase regulates the nuclear entry of the c-Jun coactivator alpha-NAC and its coactivation potency. J. Biol. Chem. 279, 43893–43899.CrossRefGoogle Scholar
  28. Rao W., Xie G., Zhang Y., Wang S., Wang Y., Zhang H. et al. 2014 OVA66, a tumor associated protein, induces oncogenic transformation of NIH3T3 cells. PLoS one 9, e85705.CrossRefGoogle Scholar
  29. Reddy V. S., Valente A. J., Delafontaine P. and Chandrasekar B. 2011 Interleukin-18/WNT1-inducible signaling pathway protein-1 signaling mediates human saphenous vein smooth muscle cell proliferation. J. Cell. Physiol. 226, 3303–3315.CrossRefGoogle Scholar
  30. Salomonis N., Hanspers K., Zambon A. C., Vranizan K., Lawlor S. C., Dahlquist K. D. et al. 2007 GenMAPP 2: new features and resources for pathway analysis. BMC Bioinf. 8, 217.CrossRefGoogle Scholar
  31. Shekaran A., Shoemaker J. T., Kavanaugh T. E., Lin A. S., LaPlaca M. C., Fan Y. et al. 2014 The effect of conditional inactivation of beta 1 integrins using twist 2 Cre, Osterix Cre and osteocalcin Cre lines on skeletal phenotype. Bone 68, 131–141.CrossRefGoogle Scholar
  32. Standaert M. L., Kanoh Y., Sajan M. P., Bandyopadhyay G. and Farese R. V. 2002 Cbl, IRS-1, and IRS-2 mediate effects of rosiglitazone on PI3K, PKC-lambda, and glucose transport in 3T3/L1 adipocytes. Endocrinology 143, 1705–1716.CrossRefGoogle Scholar
  33. Tannenbaum S. L., Hernandez M., Zheng D. D., Sussman D. A. and Lee D. J. 2014 Individual- and neighborhood-level predictors of mortality in Florida colorectal cancer patients. PLoS one 9, e106322.CrossRefGoogle Scholar
  34. Wang G. P. and Xu C. S. 2011b Alterations in DNA repair gene expression and their possible regulation in rat-liver regeneration. Genet. Mol. Biol. 34, 304–309.CrossRefGoogle Scholar
  35. Wang C., Mayer J. A., Mazumdar A., Fertuck K., Kim H., Brown M. et al. 2011a Estrogen induces c-myc gene expression via an upstream enhancer activated by the estrogen receptor and the AP-1 transcription factor. Mol. Endocrinol. 25, 1527–1538.CrossRefGoogle Scholar
  36. Wu C. and Dedhar S. 2001 Integrin-linked kinase (ILK) and its interactors: a new paradigm for the coupling of extracellular matrix to actin cytoskeleton and signaling complexes. J. Cell Biol. 155, 505–510.CrossRefGoogle Scholar
  37. Wu Q., Fan C., Chen T., Liu C., Mei W., Chen S. et al. 2013 Microwave-assisted synthesis of arene ruthenium(II) complexes that induce S-phase arrest in cancer cells by DNA damage-mediated p53 phosphorylation. Eur. J. Med. Chem. 63, 57–63.CrossRefGoogle Scholar
  38. Zhang L. and Zeng Y. 2009 Balb/c 3T3 cell transformation assay and its application to the screening of carcinogens. Wei Sheng Yan Jiu 38, 226–232.PubMedGoogle Scholar
  39. Zhao M., Sun J. and Zhao Z. 2013 TSGene: a web resource for tumor suppressor genes. Nucleic Acids Res. 41, D970–D976.CrossRefGoogle Scholar
  40. Zupancic K., Blejec A., Herman A., Veber M., Verbovsek U., Korsic M. et al. 2014 Identification of plasma biomarker candidates in glioblastoma using an antibody-array-based proteomic approach. Radiol. Oncol. 48, 257–266.CrossRefGoogle Scholar

Copyright information

© Indian Academy of Sciences 2018

Authors and Affiliations

  1. 1.State Key Laboratory Cultivation Base for Cell Differentiation RegulationHenan Normal UniversityXinxiangPeople’s Republic of China
  2. 2.Department of Histology and EmbryologyXinxiang Medical UniversityXinxiangPeople’s Republic of China
  3. 3.Henan Provincial People’s HospitalZhengzhouPeople’s Republic of China

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