Deciphering Promoter Hypermethylation of Genes Encoding for RASSF/Hippo Pathway Reveals the Poor Prognostic Factor of RASSF2 Gene Silencing in Colon Cancers
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Patients
2.2. DNA Extraction and Methylation-Specific PCR Assay
2.3. Statistical Analysis
3. Results
3.1. Clinical and Histo-Prognostic Population Characteristics
3.2. MS-PCR Results and Impact on Survival
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Ashktorab, H.; Schäffer, A.A.; Daremipouran, M.; Smoot, D.T.; Lee, E.; Brim, H. Distinct Genetic Alterations in Colorectal Cancer. PLoS ONE 2010, 5, e8879. [Google Scholar] [CrossRef] [PubMed]
- Armaghany, T.; Wilson, J.D.; Chu, Q.; Mills, G. Genetic Alterations in Colorectal Cancer. Gastrointest. Cancer Res. 2012, 5, 19–27. [Google Scholar]
- Toyota, M.; Ahuja, N.; Ohe-Toyota, M.; Herman, J.G.; Baylin, S.B.; Issa, J.-P.J. CpG Island Methylator Phenotype in Colorectal Cancer. Proc. Natl. Acad. Sci. USA 1999, 96, 8681–8686. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Weisenberger, D.J.; Siegmund, K.D.; Campan, M.; Young, J.; Long, T.I.; Faasse, M.A.; Kang, G.H.; Widschwendter, M.; Weener, D.; Buchanan, D.; et al. CpG Island Methylator Phenotype Underlies Sporadic Microsatellite Instability and Is Tightly Associated with BRAF Mutation in Colorectal Cancer. Nat. Genet. 2006, 38, 787–793. [Google Scholar] [CrossRef] [PubMed]
- Cancer Colorectal-Santé Publique France. Available online: https://www.santepubliquefrance.fr/maladies-et-traumatismes/cancers/cancer-du-colon-rectum (accessed on 11 February 2021).
- Global Cancer Observatory. Cancer Today. GLOBOCAN. 2018. Available online: https://gco.iarc.fr/today/online-analysis-multi-bars?v=2018&mode=cancer&mode_population=countries&population=900&populations=900&key=total&sex=0&cancer=39&type=0&statistic=5&prevalence=0&population_group=0&ages_group%5B%5D=0&ages_group%5B%5D=17&nb_items=1 (accessed on 13 February 2020).
- Avruch, J.; Zhou, D.; Fitamant, J.; Bardeesy, N.; Mou, F.; Barrufet, L.R. Protein Kinases of the Hippo Pathway: Regulation and Substrates. Semin. Cell Dev. Biol. 2012, 23, 770–784. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Hergovich, A. The Roles of NDR Protein Kinases in Hippo Signalling. Genes 2016, 7, 21. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Dupont, S.; Morsut, L.; Aragona, M.; Enzo, E.; Giulitti, S.; Cordenonsi, M.; Zanconato, F.; Le Digabel, J.; Forcato, M.; Bicciato, S.; et al. Role of YAP/TAZ in Mechanotransduction. Nature 2011, 474, 179–183. [Google Scholar] [CrossRef]
- Praskova, M.; Khoklatchev, A.; Ortiz-Vega, S.; Avruch, J. Regulation of the MST1 Kinase by Autophosphorylation, by the Growth Inhibitory Proteins, RASSF1 and NORE1, and by Ras. Biochem. J. 2004, 381, 453–462. [Google Scholar] [CrossRef]
- Takahashi, Y.; Miyoshi, Y.; Takahata, C.; Irahara, N.; Taguchi, T.; Tamaki, Y.; Noguchi, S. Down-Regulation of LATS1 and LATS2 MRNA Expression by Promoter Hypermethylation and Its Association with Biologically Aggressive Phenotype in Human Breast Cancers. Clin. Cancer Res. 2005, 11, 1380–1385. [Google Scholar] [CrossRef] [Green Version]
- Zhang, L.; Tang, F.; Terracciano, L.; Hynx, D.; Kohler, R.; Bichet, S.; Hess, D.; Cron, P.; Hemmings, B.A.; Hergovich, A.; et al. NDR Functions as a Physiological YAP1 Kinase in the Intestinal Epithelium. Curr. Biol. 2015, 25, 296–305. [Google Scholar] [CrossRef] [Green Version]
- Dubois, F.; Keller, M.; Calvayrac, O.; Soncin, F.; Hoa, L.; Hergovich, A.; Parrini, M.-C.; Mazières, J.; Vaisse-Lesteven, M.; Camonis, J.; et al. RASSF1A Suppresses the Invasion and Metastatic Potential of Human Non-Small Cell Lung Cancer Cells by Inhibiting YAP Activation through the GEF-H1/RhoB Pathway. Cancer Res. 2016, 76, 1627–1640. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Cooper, W.N.; Hesson, L.B.; Matallanas, D.; Dallol, A.; von Kriegsheim, A.; Ward, R.; Kolch, W.; Latif, F. RASSF2 Associates with and Stabilizes the Proapoptotic Kinase MST2. Oncogene 2009, 28, 2988–2998. [Google Scholar] [CrossRef] [Green Version]
- Song, H.; Oh, S.; Oh, H.J.; Lim, D.-S. Role of the Tumor Suppressor RASSF2 in Regulation of MST1 Kinase Activity. Biochem. Biophys. Res. Commun. 2010, 391, 969–973. [Google Scholar] [CrossRef]
- Zheng, Y.-B.; Xiao, K.; Xiao, G.-C.; Tong, S.-L.; Ding, Y.; Wang, Q.-S.; Li, S.-B.; Hao, Z.-N. MicroRNA-103 Promotes Tumor Growth and Metastasis in Colorectal Cancer by Directly Targeting LATS2. Oncol. Lett. 2016, 12, 2194–2200. [Google Scholar] [CrossRef] [Green Version]
- Sun, Z.-Q.; Shi, K.; Zhou, Q.-B.; Zeng, X.-Y.; Liu, J.; Yang, S.-X.; Wang, Q.-S.; Li, Z.; Wang, G.-X.; Song, J.-M.; et al. MiR-590-3p Promotes Proliferation and Metastasis of Colorectal Cancer via Hippo Pathway. Oncotarget 2017, 8, 58061–58071. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Schagdarsurengin, U.; Gimm, O.; Hoang-Vu, C.; Dralle, H.; Pfeifer, G.P.; Dammann, R. Frequent Epigenetic Silencing of the CpG Island Promoter of RASSF1A in Thyroid Carcinoma. Cancer Res. 2002, 62, 3698–3701. [Google Scholar] [PubMed]
- Akino, K.; Toyota, M.; Suzuki, H.; Mita, H.; Sasaki, Y.; Ohe-Toyota, M.; Issa, J.-P.J.; Hinoda, Y.; Imai, K.; Tokino, T. The Ras Effector RASSF2 Is a Novel Tumor-Suppressor Gene in Human Colorectal Cancer. Gastroenterology 2005, 129, 156–169. [Google Scholar] [CrossRef]
- Brenner, H.; Chen, C. The Colorectal Cancer Epidemic: Challenges and Opportunities for Primary, Secondary and Tertiary Prevention. Br. J. Cancer 2018, 119, 785–792. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Siegel, R.; Desantis, C.; Jemal, A. Colorectal Cancer Statistics, 2014. CA Cancer J. Clin. 2014, 64, 104–117. [Google Scholar] [CrossRef] [PubMed]
- Loree, J.M.; Pereira, A.A.L.; Lam, M.; Willauer, A.N.; Raghav, K.; Dasari, A.; Morris, V.K.; Advani, S.; Menter, D.G.; Eng, C.; et al. Classifying Colorectal Cancer by Tumor Location Rather than Sidedness Highlights a Continuum in Mutation Profiles and Consensus Molecular Subtypes. Clin. Cancer Res. 2018, 24, 1062–1072. [Google Scholar] [CrossRef] [Green Version]
- Connell, L.C.; Mota, J.M.; Braghiroli, M.I.; Hoff, P.M. The Rising Incidence of Younger Patients with Colorectal Cancer: Questions About Screening, Biology, and Treatment. Curr. Treat. Options Oncol. 2017, 18, 23. [Google Scholar] [CrossRef] [PubMed]
- Yahagi, M.; Okabayashi, K.; Hasegawa, H.; Tsuruta, M.; Kitagawa, Y. The Worse Prognosis of Right-Sided Compared with Left-Sided Colon Cancers: A Systematic Review and Meta-Analysis. J. Gastrointest. Surg. 2016, 20, 648–655. [Google Scholar] [CrossRef] [PubMed]
- Lee, G.H.; Malietzis, G.; Askari, A.; Bernardo, D.; Al-Hassi, H.O.; Clark, S.K. Is Right-Sided Colon Cancer Different to Left-Sided Colorectal Cancer?—A Systematic Review. Eur. J. Surg. Oncol. 2015, 41, 300–308. [Google Scholar] [CrossRef]
- Hesson, L.B.; Wilson, R.; Morton, D.; Adams, C.; Walker, M.; Maher, E.R.; Latif, F. CpG Island Promoter Hypermethylation of a Novel Ras-Effector Gene RASSF2A Is an Early Event in Colon Carcinogenesis and Correlates Inversely with K-Ras Mutations. Oncogene 2005, 24, 3987–3994. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Seidel, C.; Schagdarsurengin, U.; Blümke, K.; Würl, P.; Pfeifer, G.P.; Hauptmann, S.; Taubert, H.; Dammann, R. Frequent Hypermethylation of MST1 and MST2 in Soft Tissue Sarcoma. Mol. Carcinog. 2007, 46, 865–871. [Google Scholar] [CrossRef] [PubMed]
- Park, H.-W.; Kang, H.C.; Kim, I.-J.; Jang, S.-G.; Kim, K.; Yoon, H.-J.; Jeong, S.-Y.; Park, J.-G. Correlation between Hypermethylation of the RASSF2A Promoter and K-Ras/BRAF Mutations in Microsatellite-Stable Colorectal Cancers. Int. J. Cancer 2007, 120, 7–12. [Google Scholar] [CrossRef] [PubMed]
- Expression of RASSF2 in Colorectal Cancer—The Human Protein Atlas. Available online: https://www.proteinatlas.org/ENSG00000101265-RASSF2/pathology/colorectal+cancer/COAD (accessed on 19 April 2021).
- Luo, D.; Ye, T.; Li, T.-Q.; Tang, P.; Min, S.-D.; Zhao, G.-F.; Huang, H.; Chang, J.; Wang, Y.; Lv, L.; et al. Ectopic Expression of RASSF2 and Its Prognostic Role for Gastric Adenocarcinoma Patients. Exp. Ther. Med. 2012, 3, 391–396. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Aydin, D.; Bilici, A.; Kayahan, S.; Yavuzer, D.; Basar, M.; Aliustaoglu, M. Prognostic Importance of RASSF2 Expression in Patients with Gastric Cancer Who Had Undergone Radical Gastrectomy. Clin. Transl. Oncol. 2016, 18, 608–616. [Google Scholar] [CrossRef]
- Carter, J.V.; O’Brien, S.J.; Burton, J.F.; Oxford, B.G.; Stephen, V.; Hallion, J.; Bishop, C.; Galbraith, N.J.; Eichenberger, M.R.; Sarojini, H.; et al. The MicroRNA-200 Family Acts as an Oncogene in Colorectal Cancer by Inhibiting the Tumor Suppressor RASSF2. Oncol. Lett. 2019, 18, 3994–4007. [Google Scholar] [CrossRef]
- Hu, F.; Chen, L.; Bi, M.-Y.; Zheng, L.; He, J.-X.; Huang, Y.-Z.; Zhang, Y.; Zhang, X.-L.; Guo, Q.; Luo, Y.; et al. Potential of RASSF1A Promoter Methylation as a Biomarker for Colorectal Cancer: Meta-Analysis and TCGA Analysis. Pathol. Res. Pract. 2020, 216, 153009. [Google Scholar] [CrossRef]
- Dubois, F.; Bergot, E.; Zalcman, G.; Levallet, G. RASSF1A, Puppeteer of Cellular Homeostasis, Fights Tumorigenesis, and Metastasis-an Updated Review. Cell Death Dis. 2019, 10, 928. [Google Scholar] [CrossRef] [PubMed]
- Jiang, Y.; Cui, L.; Chen, W.; Shen, S.; Ding, L. The Prognostic Role of RASSF1A Promoter Methylation in Breast Cancer: A Meta-Analysis of Published Data. PLoS ONE 2012, 7, e36780. [Google Scholar] [CrossRef]
- Cao, D.; Chen, Y.; Tang, Y.; Peng, X.-C.; Dong, H.; Li, L.-H.; Cheng, K.; Ge, J.; Liu, J.-Y. Loss of RASSF1A Expression in Colorectal Cancer and Its Association with K-Ras Status. Biomed. Res. Int. 2013, 2013, 976765. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Sun, X.; Yuan, W.; Hao, F.; Zhuang, W. Promoter Methylation of RASSF1A Indicates Prognosis for Patients with Stage II and III Colorectal Cancer Treated with Oxaliplatin-Based Chemotherapy. Med. Sci. Monit. 2017, 23, 5389–5395. [Google Scholar] [CrossRef]
- Minoo, P.; Zlobec, I.; Baker, K.; Tornillo, L.; Terracciano, L.; Jass, J.R.; Lugli, A. Prognostic Significance of Mammalian Sterile20-like Kinase 1 in Colorectal Cancer. Mod. Pathol. 2007, 20, 331–338. [Google Scholar] [CrossRef] [Green Version]
- Maille, E.; Brosseau, S.; Hanoux, V.; Creveuil, C.; Danel, C.; Bergot, E.; Scherpereel, A.; Mazières, J.; Margery, J.; Greillier, L.; et al. MST1/Hippo Promoter Gene Methylation Predicts Poor Survival in Patients with Malignant Pleural Mesothelioma in the IFCT-GFPC-0701 MAPS Phase 3 Trial. Br. J. Cancer 2019, 120, 387–397. [Google Scholar] [CrossRef] [Green Version]
- Guo, Z.; Li, G.; Bian, E.; Ma, C.-C.; Wan, J.; Zhao, B. TGF-β-Mediated Repression of MST1 by DNMT1 Promotes Glioma Malignancy. Biomed. Pharmacother. 2017, 94, 774–780. [Google Scholar] [CrossRef]
- Expression of LATS2 in Colorectal Cancer—The Human Protein Atlas. Available online: https://www.proteinatlas.org/ENSG00000150457-LATS2/pathology/colorectal+cancer/COAD (accessed on 22 May 2021).
- Peng, H.; Pan, X.; Su, Q.; Zhu, L.-S.; Ma, G.-D. MiR-372-3p Promotes Tumor Progression by Targeting LATS2 in Colorectal Cancer. Eur. Rev. Med. Pharmacol. Sci. 2019, 23, 8332–8344. [Google Scholar] [CrossRef] [PubMed]
- Wierzbicki, P.M.; Adrych, K.; Kartanowicz, D.; Stanislawowski, M.; Kowalczyk, A.; Godlewski, J.; Skwierz-Bogdanska, I.; Celinski, K.; Gach, T.; Kulig, J.; et al. Underexpression of LATS1 TSG in Colorectal Cancer Is Associated with Promoter Hypermethylation. World J. Gastroenterol. 2013, 19, 4363–4373. [Google Scholar] [CrossRef]
- Zhang, Y.; Hu, C.-F.; Chen, J.; Yan, L.-X.; Zeng, Y.-X.; Shao, J.-Y. LATS2 Is De-Methylated and Overexpressed in Nasopharyngeal Carcinoma and Predicts Poor Prognosis. BMC Cancer 2010, 10, 538. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Chen, K.-H.; He, J.; Wang, D.-L.; Cao, J.-J.; Li, M.-C.; Zhao, X.-M.; Sheng, X.; Li, W.-B.; Liu, W.-J. Methylation-associated Inactivation of LATS1 and Its Effect on Demethylation or Overexpression on YAP and Cell Biological Function in Human Renal Cell Carcinoma. Int. J. Oncol. 2014, 45, 2511–2521. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Malik, S.A.; Khan, M.S.; Dar, M.; Hussain, M.U.; Shah, M.A.; Shafi, S.M.; Mudassar, S. Molecular Alterations and Expression Dynamics of LATS1 and LATS2 Genes in Non-Small-Cell Lung Carcinoma. Pathol. Oncol. Res. 2018, 24, 207–214. [Google Scholar] [CrossRef] [PubMed]
- Kim, E.; Ahn, B.; Oh, H.; Lee, Y.J.; Lee, J.H.; Lee, Y.; Kim, C.H.; Chae, Y.-S.; Kim, J.Y. High Yes-Associated Protein 1 with Concomitant Negative LATS1/2 Expression Is Associated with Poor Prognosis of Advanced Gastric Cancer. Pathology 2019, 51, 261–267. [Google Scholar] [CrossRef]
- Levallet, G.; Creveuil, C.; Bekaert, L.; Péres, E.; Planchard, G.; Lecot-Cotigny, S.; Guillamo, J.-S.; Emery, E.; Zalcman, G.; Lechapt-Zalcman, E. Promoter Hypermethylation of Genes Encoding for RASSF/Hippo Pathway Members Reveals Specific Alteration Pattern in Diffuse Gliomas. J. Mol. Diagn. 2019, 21, 695–704. [Google Scholar] [CrossRef]
Clinical Characteristics | All Patients N = 229 | |
---|---|---|
N | % | |
Age (years old) | 71 (27–99) | |
Sex (female/male) | 109/120 | 47.6/52.4 |
Location (right/left) | 119/110 | 52.0/48.0 |
Tumoral size (cm) | 5.05 (0.4–16) | |
Clinical stage | 21/81/87/40 | 9.1/35.4/38.0/17.5 |
I/II/III/IV | ||
Intestinal occlusion | 67 | 29.2 |
Synchronous metastasis (no/hepatic/extrahepatic) | 188/35/6 | 82.1/15.3/2.6 |
Recurrence | ||
No | 180 | 78,6 |
Local/Hepatic/Extrahepatic | 18/19/8 | 7.9/8.3/3.5 |
Death | 96 | 40.6 |
Histological Characteristics | All Patients N = 229 | |
---|---|---|
N | % | |
Differentiation Well/Moderately/Poor Mucinous | 101/76/23 29 | 44.1/33.2/10.0 12.7 |
Stage T 1/2/3/4a/4b | 7/18/140/51/2 | 3.0/7.8/61.1/22.2/0.9 |
Stage N 0/1a/1b/1c/2 | 113/34/37/9/36 | 49.3/14.9/16.2/3.9/15.7 |
Emboli | 96 | 41.9 |
MMR (pMMR/dMMR/NA 1) | 24/10/195 | 10.5/4.4/85.1 |
RAS (no, mutated/NA 1) | 46/34/149 | 20.1/14.8/65.1 |
BRAF (no, mutated/NA 1) | 69/6/154 | 30.1/2.6/67.3 |
RASSF2 hypermethylation | 199 | 86.9 |
Clinical and Histological | Univariate Model | ||
---|---|---|---|
Characteristics | HR | IC 95% | p |
Age (<70 years old) | 2.07 | [0.95–4.50] | 0.07 |
Sex (male) | 1.98 | [0.88–4.44] | 0.10 |
Location (right) | 0.57 | [0.26–1.25] | 0.16 |
Synchronous metastasis (no metastasis) | 1.07 | [0.38–2.98] | 0.90 |
Tumoral deposit | 2.00 | [0.73–5.50] | 0.18 |
Lymphatic tumor emboli | 0.94 | [0.43–2.03] | 0.86 |
Budding | 1.25 | [0.48–3.25] | 0.64 |
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Riffet, M.; Eid, Y.; Faisant, M.; Fohlen, A.; Menahem, B.; Alves, A.; Dubois, F.; Levallet, G.; Bazille, C. Deciphering Promoter Hypermethylation of Genes Encoding for RASSF/Hippo Pathway Reveals the Poor Prognostic Factor of RASSF2 Gene Silencing in Colon Cancers. Cancers 2021, 13, 5957. https://doi.org/10.3390/cancers13235957
Riffet M, Eid Y, Faisant M, Fohlen A, Menahem B, Alves A, Dubois F, Levallet G, Bazille C. Deciphering Promoter Hypermethylation of Genes Encoding for RASSF/Hippo Pathway Reveals the Poor Prognostic Factor of RASSF2 Gene Silencing in Colon Cancers. Cancers. 2021; 13(23):5957. https://doi.org/10.3390/cancers13235957
Chicago/Turabian StyleRiffet, Marc, Yassine Eid, Maxime Faisant, Audrey Fohlen, Benjamin Menahem, Arnaud Alves, Fatéméh Dubois, Guénaelle Levallet, and Céline Bazille. 2021. "Deciphering Promoter Hypermethylation of Genes Encoding for RASSF/Hippo Pathway Reveals the Poor Prognostic Factor of RASSF2 Gene Silencing in Colon Cancers" Cancers 13, no. 23: 5957. https://doi.org/10.3390/cancers13235957