JBM  Vol.7 No.5 , May 2019
Association of H3K4me3 and CBX7 Levels to the Down-Regulation of GKN1 Gene Expression in Gastric Cancer Cells
Abstract: Gastrokine 1 (GKN1) is a highly secreted gastric mucosal protein in normal individuals but strongly down-regulated or totally absent in gastric cancer subjects. An epigenetic mechanism might be responsible for GKN1 gene silencing probably through the activity of a transcription factor in association with the enzymes SUV39H1 and HDACs on the GKN1 promoter. In fact, compared to non-tumor tissues, a high increase of H3K9me3 level was observed in the corresponding tumor ones. Because H3K4me3 seems to be a possible epigenetic mark for active euchromatin, we try to verify the H3K4me3 level on the GKN1 promoter in gastric cancer tumor specimens. In addition, we also attempt to highlight if CBX7 could be the possible regulatory transcription factor correlated to GKN1 gene promoter. Therefore, we evaluated if the CBX7 expression levels could be associated with GKN1 down-regulation in gastric cancer. To this purpose, 2 pairs of non-tumor and tumor surgical specimens from patients with gastric cancer were analyzed for H3K4me3 by chromatin immunoprecipitation (ChiP) assays, and 9 pairs were instead analyzed by Western blotting for GKN1, and CBX7 expression levels, respectively. The results suggested that the observed increase of H3K4me3 in tumor samples was not in agreement with its proposed function whereas the expression of CBX7 was not associated with the down-regulation of GKN1. In particular, the expression levels of CBX7 in tumor samples might suggest a survival role in gastric cancer.
Cite this paper: Di Stadio, C. , Altieri, F. , Federico, A. , Miselli, G. , Grillo, M. , Rippa, E. and Arcari, P. (2019) Association of H3K4me3 and CBX7 Levels to the Down-Regulation of GKN1 Gene Expression in Gastric Cancer Cells. Journal of Biosciences and Medicines, 7, 146-156. doi: 10.4236/jbm.2019.75016.

[1]   Oien, K.A., McGregor, F., Butler, S., Ferrier, R.K., Downie, I., Bryce, S., Burns, S. and Keith, W.N. (2004) Gastrokine 1 Is Abundantly and Specifically Expressed in Superficial Gastric Epithelium, Down-Regulated in Gastric Carcinoma, and Shows High Evolutionary Conservation. The Journal of Pathology, 203, 789-797.

[2]   Nardone, G., Martin, G., Rocco, A., Rippa, E., La Monica, G., Caruso, F. and Arcari, P. (2008) Molecular Expression of Gastrokine 1 in Normal Mucosa and in Helicobacter pylori-Related Preneoplastic and Neoplastic Gastric Lesions. Cancer Biology and Therapy, 7, 1890-1895.

[3]   Rippa, E., La Monica, G., Allocca, R., Romano, M.F., De Palma, M. and Arcari, P. (2011) Overexpression of Gastrokine 1 in Gastric Cancer Cells Induces Fas-Mediated Apoptosis. Journal of Cellular Physiology, 226, 2571-2578.

[4]   Pavone, L.M.N., Del Vecchio, P., Mallardo, P., Altieri, F., De Pasquale, V., Rea, S., Martucci, N.M., Di Stadio, C.S., Pucci, P., Flagiello, A., Masullo, M., Arcari, P. and Rippa, E. (2013) Structural Characterization and Biological Properties of Human Gastrokine 1. Molecular BioSystems, 9, 412-421.

[5]   Altieri, F., Di Stadio, C.S., Federico, A., Miselli, G., De Palma, M., Rippa, E. and Arcari, P. (2017) Epigenetic Alterations of Gastrokine 1 Gene Expression in Gastric Cancer. Oncotarget, 810, 16899-16911.

[6]   Bernstein, B.E., Humphrey, E.L., Erlich, R.L., Schneider, R., Bouman, P., Liu, J.S., Kouzarides, T. and Schreiber, S.L. (2002) Methylation of Histone H3 Lys 4 in Coding Regions of Active Genes. Proceedings of the National Academy of Sciences of the United States of America, 99, 8695-8700.

[7]   Schneider, R., Bannister, A.J., Myers, F.A., Thorne, A.W., Crane-Robinson, C. and Kouzarides, T. (2004) Histone H3 Lysine 4 Methylation Patterns in Higher Eukaryotic Genes. Nature Cell Biology, 6, 73-77.

[8]   Bannister, A.J., Zegerman, P., Partridge, J.F., Miska, E.A., Thomas, J.O., Allshire, R.C. and Kouzarides, T. (2001) Selective Recognition of Methylated Lysine 9 on Histone H3 by the HP1 Chromo Domain. Nature, 410, 120-124.

[9]   Stewart, M.D., Li, J. and Wong, J. (2005) Relationship between Histone H3 Lysine 9 Methylation, Transcription Repression, and Heterochromatin Protein 1 Recruitment. Molecular and Cellular Biology, 25, 2525-2538.

[10]   Di Croce, L. and Helin, K. (2013) Transcriptional Regulation by Polycomb Group Proteins. Nature Structural and Molecular Biology, 20, 1147-1155.

[11]   Pallante, P., Federico, A., Berlingieri, M.T., Bianco, M., Ferraro, A., Forzati, F., Iaccarino, A., Russo, M., Pierantoni, G.M., Leone, V., Sacchetti, S., Troncone, G., Santoro, M. and Fusco, A. (2008) Loss of the CBX7 Gene Expression Correlates with a Highly Malignant Phenotype in Thyroid Cancer. Cancer Research, 68, 6770-6778.

[12]   Pallante, P., Terracciano, L., Carafa, V., Schneider, S., Zlobec, I., Lugli, A., Bianco, M., Ferraro, A., Sacchetti, S., Troncone, G., Fusco, A. and Tornillo, L. (2010) The Loss of the CBX7 Gene Expression Represents an Adverse Prognostic Marker for Survival of Colon Carcinoma Patients. European Journal of Cancer, 46, 2304-2313.

[13]   Karamitopoulou, E., Pallante, P., Zlobec, I., Tornillo, L., Carafa, V., Schaffner, T., Borner, M., Diamantis, I., Esposito, F., Brunner, T., Zimmermann, A., Federico, A., Terracciano, L. and Fusco, A. (2010) Loss of the CBX7 Protein Expression Correlates with a More Aggressive Phenotype in Pancreatic Cancer. European Journal of Cancer, 46, 1438-1444.

[14]   Zhang, X.W., Zhang, L., Qin, W., Yao, X.H., Zheng, L.Z., Liu, X., Li, J. and Guo, W.J. (2010) Oncogenic Role of the Chromobox Protein CBX7 in Gastric Cancer. Journal of Experimental and Clinical Cancer Research, 29, Article ID: 4320.

[15]   Forzati, F., Federico, A., Pallante, P., Fedele, M. and Fusco, A. (2012) Tumor Suppressor Activity of CBX7 in Lung Carcinogenesis. Cell Cycle, 11, 1888-1891.

[16]   Ni, S.-J., Zhao, L.-Q., Wang, X.-F., Wu, Z.-H., Hua, R.-X., Wan, C.-H., Zhang, J.-Y., Zhang, X.-W., Huang, M.-Z., Gan, L., Sun, H.-L., Dimri, G.P. and Guo, W.-J. (2018) CBX7 Regulates Stem Cell-Like Properties of Gastric Cancer Cells via p16 and AKT-NF-κBmiR-21 Pathways. Journal of Hematology and Oncology, 11, Article ID: 1276.

[17]   Di Stadio, C.S., Altieri, F., Federico, A., Miselli, G., Niglio, A., De Palma, M., Rippa, E. and Arcari, P. (2017) Down-Regulation of GKN1 in Gastric Cancer Is Not Associated with the RUNX3 Expression. Journal of Biosciences and Medicines, 5, 1-11.

[18]   Di Stadio, C.S., Altieri, F., Miselli, G., Elce, A., Severino, V., Chambery, A., Quagliariello, V., Villano, V., de Dominicis, G., Rippa, E. and Arcari, P. (2016) AMP18 Interacts with the Anion Exchanger SLC26A3 and Enhances Its Expression in Gastric Cancer Cells. Biochimie, 121, 151-160.

[19]   Edge, S.B. and Compton, C.C. (2010) The American Joint Committee on Cancer: The 7th Edition of the AJCC Cancer Staging Manual and the Future of TNM. Annals of Surgical Oncology, 17, 1471-1474.

[20]   Tsunoda, T. and Takagi, T. (1999) Estimating Transcription Factor Bindability on DNA. Bioinformatics, 15, 622-630.

[21]   Lee, C. and Huang, C.H. (2013) LASAGNA-Search: An Integrated Web Tool for Transcription Factor Binding Site Search and Visualization. BioTechniques, 54, 141-153.

[22]   Messeguer, X., Escudero, R., Farré, D., Nunez, O., Martínez, J. and Albà, M.M. (2002) PROMO: Detection of Known Transcription Regulatory Elements Using Species-Tailored Searches. Bioinformatics, 18, 333-334.

[23]   Farré, D., Roset, R., Huerta, M., Adsuara, J.E., Roselló, L., Albà, M.M. and Messeguer, X. (2003) Identification of Patterns in Biological Sequences at the ALGGEN Server: PROMO and MALGEN. Nucleic Acids Research, 31, 3651-3653.

[24]   Cruz, C., Rosa, D.M., Krueger, C., Gao, Q., Horkai, D., King. M., Field, M. and Houseley, J. (2018) Trimethylation of Histone H3 Lysine 4 Facilitates Gene Expression in Ageing Cells. eLife, 7, e34081.

[25]   Santos-Rosa, H., Schneider, R., Bannister, A.J., Sherriff, J., Bernstein, B.E., Emre, N.C., Schreiber, S.L., Mellor, J. and Kouzarides, T. (2002) Active Genes Are Tri-Methylated at K4 of Histone H3. Nature, 419, 407-411.

[26]   Howe, F.S., Fischl, H., Murray, S.C. and Mellor, J. (2016) Is H3K4me3 Instructive for Transcription Activation? Bioessays, 39, 1-12.