Abstrakt

Aim: The activation of canonical Wnt pathway is etiologically associated with the development of colorectal cancers. There are many possible molecular targets for the therapeutic abrogation of Wnt/β-catenin signaling. The aim of this study was to select the best molecular targets for the attenuation of β-catenin-dependent gene expression in colorectal cancer cell lines. Material/Methods: An siRNA screen was used for the selection of the best molecular targets for the down-regulation of TCF/LEF-dependent GFP expression in HCT116 cells. The level of the expression of β-catenin target genes was analyzed by qPCR. The effect of the tested chemicals on cell migration, cell cycle and apoptosis was assessed by the wound healing assay, flow cytometric analysis of propidium iodide stained cells and flow cytometric analysis of the activity of caspases-3/7, respectively. Results: Of the forty three genes which were tested in the screening stage, eight (KDM6A, KDM1A, PORCN, KDM4C, CARM1, DVL1, CBP, KMT2A) were selected as most promising. Small molecule inhibitors of these proteins (GSK-J4, GSK-LSD1, IWP-2, ML324, MS049, Dvl-PDZ Domain Inhibitor II, PRI-724, MM-102) were further used. The inhibitors of Porcupine (IWP-2) and CBP (PRI-724) were most effective in the down-regulation of the expression of β-catenin target genes and the induction of apoptosis in HCT116 cells, but showed weaker effects in DLD-1 cells. Conclusions: The inhibition of CBP and Porcupine is the most effective in attenuating canonical Wnt signaling in colon cancer cells. Future studies should determine which factors affect the sensitivity towards these promising anti-cancer agents.

Przypisy

• 1. Alsaqer S.F., Tashkandi M.M., Kartha V.K., Yang YT., Alkheriji Y.,Salama A., Varelas X., Kukuruzinska M., Monti S., Bais M.V.: Inhibitionof LSD1 epigenetically attenuates oral cancer growth and metastasis.Oncotarget, 2017; 8: 73372–73386
  Google Scholar
• 2. Ayadi M., Bouygues A., Ouaret D., Ferrand N., Chouaib S., ThieryJ.P., Muchardt C., Sabbah M., Larsen A.K.: Chronic chemotherapeuticstress promotes evolution of stemness and WNT/beta-cateninsignaling in colorectal cancer cells: implications for clinical use ofWNT-signaling inhibitors. Oncotarget, 2015; 6: 18518–18533
  Google Scholar
• 3. Berry W.L., Janknecht R.: KDM4/JMJD2 histone demethylases:epigenetic regulators in cancer cells. Cancer Res., 2013; 73: 2936–2942
  Google Scholar
• 4. Berry W.L., Kim T.D., Janknecht R.: Stimulation of β-catenin andcolon cancer cell growth by the KDM4B histone demethylase. Int. J.Oncol., 2014; 44: 1341–1348 5 Covey T.M., Kaur S., Tan Ong T., Proffitt K.D., Wu Y., Tan P., VirshupD.M.: PORCN moonlights in a Wnt-independent pathway that regulatescancer cell proliferation. PLoS One, 2012; 7: e34532
  Google Scholar
• 5. expression. J. Cancer Res. Clin. Oncol., 2015; 141: 11–21
  Google Scholar
• 6. Gaddis M., Gerrard D., Frietze S., Farnham P.J.: Altering cancertranscriptomes using epigenomic inhibitors. Epigenetics Chromatin,2015; 8: 9
  Google Scholar
• 7. Grandy D., Shan J., Zhang X., Rao S., Akunuru S., Li H., Zhang Y.,Alpatov I., Zhang X.A., Lang R.A., Shi D.L., Zheng J.J.: Discovery andcharacterization of a small molecule inhibitor of the PDZ domain ofdishevelled. J. Biol. Chem., 2009; 284: 16256–16263
  Google Scholar
• 8. Hanson A.J., Wallace H.A., Freeman T.J., Beauchamp R.D., Lee L.A.,Lee E.: XIAP monoubiquitylates Groucho/TLE to promote canonicalWnt signaling. Mol. Cell. 2012; 45: 619–628
  Google Scholar
• 9. Hsu H.C., Liu Y.S., Tseng K.C., Yang T.S., Yeh C.Y., You J.F., HungH.Y., Chen S.J., Chen H.C.: CBB1003, a lysine-specific demethylase 1inhibitor, suppresses colorectal cancer cells growth through downregulationof leucine-rich repeat-containing G-protein-coupled receptor
  Google Scholar
• 10. Huang Z., Li S., Song W., Li X., Li Q., Zhang Z., Han Y., ZhangX., Miao S., Du R., Wang L.: Lysine-specific demethylase 1 (LSD1/KDM1A) contributes to colorectal tumorigenesis via activation ofthe Wnt/β-catenin pathway by down-regulating Dickkopf-1 (DKK1).PLoS One, 2013; 8: e70077
  Google Scholar
• 11. Iorio F., Knijnenburg T.A., Vis D.J., Bignell G.R., Menden M.P.,Schubert M., Aben N., Goncalves E., Barthorpe S., Lightfoot H., CokelaerT., Greninger P., van Dyk E., Chang H., de Silva H., et al.: A landscapeof pharmacogenomic interactions in cancer. Cell, 2016; 166: 740–754
  Google Scholar
• 12. Kim T.D., Fuchs J.R., Schwartz E., Abdelhamid D., Etter J., BerryW.L., Li C., Ihnat M.A., Li P.K., Janknecht R.: Pro-growth role of theJMJD2C histone demethylase in HCT-116 colon cancer cells and identificationof curcuminoids as JMJD2 inhibitors. Am. J. Transl. Res.,2014; 6: 236–247
  Google Scholar
• 13. Krishnamurthy N., Kurzrock R.: Targeting the Wnt/beta-cateninpathway in cancer: Update on effectors and inhibitors. Cancer Treat.Rev., 2018; 62: 50–60
  Google Scholar
• 14. Mo M.L., Li M.R., Chen Z., Liu X.W., Sheng Q., Zhou H.M.: Inhibitionof the Wnt palmitoyltransferase porcupine suppresses cellgrowth and downregulates the Wnt/β-catenin pathway in gastriccancer. Oncol. Lett., 2013; 5: 1719–1723
  Google Scholar
• 15. Nusse R., Clevers H.: Wnt/β-catenin signaling, disease, andemerging therapeutic modalities. Cell, 2017; 169: 985–999
  Google Scholar
• 16. Ou C.Y., LaBonte M.J., Manegold P.C., So A.Y., Ianculescu I., GerkeD.S., Yamamoto K.R., Ladner R.D., Kahn M., Kim J.H., Stallcup M.R.:A coactivator role of CARM1 in the dysregulation of β-catenin activityin colorectal cancer cell growth and gene expression. Mol.Cancer Res., 2011; 9: 660–670
  Google Scholar
• 17. Paluszczak J., Kleszcz R., Studzińska-Sroka E., Krajka-Kuźniak V.:Lichen-derived caperatic acid and physodic acid inhibit Wnt signalingin colorectal cancer cells. Mol. Cell. Biochem., 2018; 441: 109–124
  Google Scholar
• 18. Polakis P.: Drugging Wnt signalling in cancer. EMBO J., 2012;31: 2737–2746
  Google Scholar
• 19. Siar C.H., Nagatsuka H., Han P.P., Buery R.R., Tsujigiwa H., NakanoK., Ng K.H., Kawakami T.: Differential expression of canonicaland non-canonical Wnt ligands in ameloblastoma. J. Oral Pathol.Med., 2012; 41: 332–339
  Google Scholar
• 20. Takahashi-Yanaga F., Kahn M.: Targeting Wnt signaling: Canwe safely eradicate cancer stem cells? Clin. Cancer Res., 2010; 16:3153–3162
  Google Scholar
• 21. Urushibara S., Tsubota T., Asai R., Azumi J., Ashida K., FujiwaraY., Shiota G.: WNT/β-catenin signaling inhibitor IC-2 suppressessphere formation and sensitizes colorectal cancer cells to 5-fluorouracil.Anticancer Res., 2017; 37: 4085–4091
  Google Scholar
• 22. Verkaar F., Zaman G.J.: New avenues to target Wnt/β-cateninsignaling. Drug Discov. Today, 2011; 16: 35–41
  Google Scholar
• 23. Wend P., Fang L., Zhu Q., Schipper J.H., Loddenkemper C., KoselF., Brinkmann V., Eckert K., Hindersin S., Holland J.D., Lehr S., KahnM., Ziebold U., Birchmeier W.: Wnt/β-catenin signalling inducesMLL to create epigenetic changes in salivary gland tumours. EMBOJ., 2013; 32: 1977–1989
  Google Scholar
• 24. Wiese M., Walther N., Diederichs C., Schill F., Monecke S., SalinasG., Sturm D., Pfister S.M., Dressel R., Johnsen S.A., Kramm C.M.: Theβ-catenin/CBP-antagonist ICG-001 inhibits pediatric glioma tumorigenicityin a Wnt-independent manner. Oncotarget, 2017; 8: 27300–27313
  Google Scholar
• 25. Yamamoto S., Tateishi K., Kudo Y., Yamamoto K., Isagawa T.,Nagae G., Nakatsuka T., Asaoka Y., Ijichi H., Hirata Y., Otsuka M.,Ikenoue T., Aburatani H., Omata M., Koike K.: Histone demethylaseKDM4C regulates sphere formation by mediating the cross talk betweenWnt and Notch pathways in colonic cancer cells. Carcinogenesis,2013; 34: 2380–2388
  Google Scholar
• 26. Zhao L., Li W., Zang W., Liu Z., Xu X., Yu H., Yang Q., Jia J.: JMJD2Bpromotes epithelial-mesenchymal transition by cooperatingwith β-catenin and enhances gastric cancer metastasis. Clin. CancerRes., 2013; 19: 6419–6429
  Google Scholar

Pełna treść artykułu