INHIBITOR OF PROTEIN KINASES 1-(4-CHLOROBENZYL)-3-CHLORO-4-(3-TRIFLUOROMETHYLPHENYLAMINO)-1H-PYRROLE-2,5-DIONE INDUCES DNA DAMAGE AND APOPTOSIS IN HUMAN COLON CARCINOMA CELLS
DOI: http://dx.doi.org/10.30970/sbi.1404.636
Abstract
Background. The heterocyclic scaffolds are in the list of key structural blocks used at synthesis of novel biologically active compounds.
Materials and Methods. The present study addressed the evaluation of the mechanisms of the DNA damaging and pro-apoptotic actions in vitro of the maleimide derivative 1-(4-chlorobenzyl)-3-chloro-4-(3-trifluoromethylphenylamino)-1Н-pyrrole-2,5-dione (MI-1) targeting human colon carcinoma cells of HCT116 line. The Western-blot analysis was used to study changes in apoptosis-associated proteins, DNA comet assay under alkaline conditions was applied for evaluation of the DNA-damaging events, and Barton’s assay with diphenylamine was applied for measuring the level of DNA fragmentation in human colon carcinoma cells treated with MI-1 compound.
Results. The results of the Western-blot analysis demonstrated that MI-1 induced the apoptosis in HCT116 cells via mitochondria-dependent pathway. It activated caspase 3 via its cleavage in the treated human colon carcinoma cells. Besides, MI-1 increased the content of mitochondria-specific proteins: endonuclease G (EndoG) and the pro-apoptotic cytosolic protein protease-activating factor 1 (Apaf1). At the same time, MI-1 reduced the level of the anti-apoptotic Bcl-2 protein in HCT116 cells. The DNA comet analysis under alkaline conditions of the targeted human colon carcinoma cells of HCT116 line demonstrated that MI-1 induced DNA single-strand breaks in line with the olive tail moment of 13.2. The results of the colorimetric diphenylamine assay in HCT116 cells have shown that cell treatment with MI-1 increased the content of fragmented DNA to 14.2 %.
Conclusions. The anti-proliferative action of MI-1 in human colon carcinoma cells of HCT116 line is associated with apoptosis induction via mitochondria-dependent pathway, as well as the DNA damage through single-strand breaks and DNA fragmentation. These data suggest that the 1-(4-chlorobenzyl)-3-chloro-4-(3-trifluoromethylphenylamino)-1Н-pyrrole-2,5-dione (MI-1) might be a promising agent for suppression of growth of colon tumor cells.
Keywords
Full Text:
PDFReferences
1. An W., Lai H., Zhang Y., Liu M., Lin X., Cao S. Apoptotic pathway as the therapeutic target for anticancer traditional chinese medicines. Frontiers in Pharmacology, 2019; 10: 758. Crossref ● PubMed ● PMC ● Google Scholar | ||||
| ||||
2. Arora S., Tandon S. DNA fragmentation and cell cycle arrest: a hallmark of apoptosis induced by Ruta graveolens in human colon cancer cells. Homeopathy, 2015; 104(1): 36-47. Crossref ● PubMed ● Google Scholar | ||||
| ||||
3. Bao H., Zhang Q., Zhu Z., Xu H., Ding F., Wang M., Du S., Du Y., Yan Z. BHX, a novel pyrazoline derivative, inhibits breast cancer cell invasion by reversing the epithelial-mesenchymal transition and down-regulating Wnt/β-catenin signalling. Scientific Reports, 2017; 7(1): 9153. Crossref ● PubMed ● PMC ● Google Scholar | ||||
| ||||
4. Basnakian A.G., Apostolov E.O., Yin X., Abiri S.O., Stewart A.G., Singh A.B., Shah S.V. Endonuclease G promotes cell death of non-invasive human breast cancer cells. Experimental Cell Research, 2006; 312(20): 4139-4149. Crossref ● PubMed ● PMC ● Google Schola | ||||
| ||||
5. Campbell K.J., Tait S. Targeting BCL-2 regulated apoptosis in cancer. Open Biology, 2018; 8(5): 180002. Crossref ● PubMed ● PMC ● Google Scholar | ||||
| ||||
6. Dubinina G., Golovach S., Kozlovsky V., Tolmachov A.O., Volovenko Yu.M. Antiproliferative action of the new derivatives of l-(4-R-benzyl)-3-R1-4-(R2-phenylamino)-1H-pyrrol-2,5-dione. Journal of Organic and Pharmaceutical Chemistry, 2007; 5(1): 39-49. (In Ukrainian) Google Scholar | ||||
| ||||
7. Dubinina G.G., Chupryna O.O., Platonov M.O., Borisko P.O., Ostrovska G.V., Tolmachov A.O., Shtil A.A. In silico design of protein kinase inhibitors: successes and failures. Anti-Cancer Agents in Medicinal Chemistry, 2007; 7(2): 171-188. Crossref ● PubMed ● Google Scholar | ||||
| ||||
8. Feng N., Luo J., Guo X. Silybin suppresses cell proliferation and induces apoptosis of multiple myeloma cells via the PI3K/Akt/mTOR signaling pathway. Molecular Medicine Reports, 2016; 13: 3243-3248. Crossref ● PubMed ● Google Scholar | ||||
| ||||
9. Finiuk N., Klyuchivska O., Ivasechko I., Hreniukh V., Ostapiuk Y., Shalai Y., Panchuk R., Matiychuk V., Obushak M., Stoika R., Babsky A. Proapoptotic effects of novel thiazole derivative on human glioma cells. Anti-Cancer Drugs, 2019; 30(1): 27-37. Crossref ● PubMed ● Google Scholar | ||||
| ||||
10. Finiuk N.S., Ivasechko I.I., Klyuchivska O.Yu., Kuznietsova H.M., Rybalchenko V.K., Stoika R.S. Cytotoxic action of maleimide derivative 1-(4-Cl-benzyl)-3-chloro-4-(CF(3)-phenylamino)-1H-pyrrole-2,5-dione toward mammalian tumor cells and its capability to interact with DNA. The Ukrainian Biochemical Journal, 2020; 92(4): 55-62. Crossref | ||||
| ||||
11. Finiuk N.S., Ivasechko I.I., Klyuchivska O.Yu., Ostapiuk Yu.V., Hreniukh V.P., Shalai Ya.R., Matiychuk V.S., Obushak M.D., Babsky A.M., Stoika R.S. Apoptosis induction in human leukemia cells by novel 2-amino-5-benzylthiazole derivatives. The Ukrainian Biochemical Journal, 2019; 91(2): 29-39. Crossref ● Google Scholar | ||||
| ||||
12. Hsu C.C., Tseng L.M., Lee H.C. Role of mitochondrial dysfunction in cancer progression. Experimental Biology and Medicine, 2016; 241(12): 1281-1295. Crossref ● PubMed ● PMC ● Google Scholar | ||||
| ||||
13. Jain C.K., Majumder H.K., Roychoudhury S. Natural compounds as anticancer agents targeting DNA topoisomerases. Current Genomics, 2017; 18(1): 75-92. Crossref ● PubMed ● PMC ● Google Scholar | ||||
| ||||
14. Jiang Z.Q., Li M.H., Qin Y.M., Jiang H.Y., Zhang X., Wu M.H. Luteolin inhibits tumorigenesis and induces apoptosis of non-small cell lung cancer cells via regulation of MicroRNA-34a-5p. International Journal of Molecular Sciences, 2018; 19(2): 447. Crossref ● PubMed ● PMC ● Google Scholar | ||||
| ||||
15. Kale M., Sonwane G., Nawale R., Mourya V. Molecular modeling studies on some important anticancer heterocycles: an overview. Current Computer-Aided Drug Design, 2018; 14(3): 178-190. Crossref ● PubMed ● Google Scholar | ||||
| ||||
16. Khan M., Maryam A., Qazi J.I., Ma T. Targeting apoptosis and multiple signaling pathways with icariside II in cancer cells. International Journal of Biological Sciences, 2015; 11: 1100-1112. Crossref ● PubMed ● PMC ● Google Scholar | ||||
| ||||
17. Kuznietsova H.M., Lynchak O.V., Danylov M.O., Kotliar I.P., Rybalchenko V.K. Effect of dihydropyrrol and maleimide derivatives on the state of liver and colon in normal rats and those with colorectal carcinogenesis induced by dimethylhydrazine. The Ukrainian Biochemical Journal, 2013; 85(3): 74-84. Crossref ● PubMed ● Google Scholar | ||||
| ||||
18. Kuznietsova H.M., Yena M.S., Kotlyar I.P., Ogloblya O.V., Rybalchenko V.K. Anti-inflammatory effects of protein kinase inhibitor pyrrol derivate. The Scientific World Journal, 2016; 2016: 2145753. Crossref ● PubMed ● PMC ● Google Scholar | ||||
| ||||
19. Lang D.K., Kaur R., Arora R., Saini B., Arora S. Nitrogen containing heterocycles as anticancer agents: an overview. Anti-Cancer Agents in Medicinal Chemistry, 2020; 20(18): 2150-2168. Crossref ● PubMed ● Google Scholar | ||||
| ||||
20. Liao W., McNutt M.A., Zhu W.G. The comet assay: a sensitive method for detecting DNA damage in individual cells. Methods, 2009; 48(1): 46-53. Crossref ● PubMed ● Google Scholar | ||||
| ||||
21. Lu M., Wang Y., Zhan X. The MAPK pathway-based drug therapeutic targets in pituitary adenomas. Frontiers in Endocrinology, 2019; 10: 330. Crossref ● PubMed ● PMC ● Google Scholar | ||||
| ||||
22. Lu Y., Liu Y., Yang C. Evaluating in vitro DNA damage using comet assay. Journal of Visualized Experiments, 2017; 128: 56450. Crossref ● PubMed ● PMC ● Google Scholar | ||||
| ||||
23. Mahmood T., Yang P.C. Western blot: technique, theory, and trouble shooting. North American Journal of Medical Sciences, 2012; 4(9): 429-434. Crossref ● PubMed ● PMC ● Google Scholar | ||||
| ||||
24. Martins P., Jesus J., Santos S, Raposo L.R., Roma-Rodrigues C., Baptista P.V., Fernandes A.R. Heterocyclic anticancer compounds: recent advances and the paradigm shift towards the use of nanomedicine's tool box. Molecules, 2015; 20(9): 16852-16891. Crossref ● PubMed ● PMC ● Google Scholar | ||||
| ||||
25. McIlwain D.R., Berger T., Mak T.W. Caspase functions in cell death and disease. Cold Spring Harbor Perspectives in Biology, 2013; 5(4): a008656. Crossref ● PubMed ● PMC ● Google Scholar | ||||
| ||||
26. Mitra S., Nguyen L.N., Akter M., Park G., Choi E.H., Kaushik N.K. Impact of ROS generated by chemical, physical, and plasma techniques on cancer attenuation. Cancers, 2019; 11(7): 1030. Crossref ● PubMed ● PMC ● Google Scholar | ||||
| ||||
27. Ngoi N., Choong C., Lee J., Bellot G., Wong A., Goh B.C., Pervaiz S. Targeting mitochondrial apoptosis to overcome treatment resistance in cancer. Cancers, 2020; 12(3): 574. Crossref ● PubMed ● PMC ● Google Scholar | ||||
| ||||
28. Pat. 22204 UA. Compound of 1,4-disubstituted 5-amino-1,2-dihydropyrrole-3-one having anticancer activity. Dubinina G.G., Volovenko Yu.M. Publ. 25.04.2007. | ||||
| ||||
29. Pfeffer C.M., Singh A.T.K. Apoptosis: A target for anticancer therapy. International Journal of Molecular Sciences, 2018; 19(2): 448. Crossref ● PubMed ● PMC ● Google Scholar | ||||
| ||||
30. Pistritto G., Trisciuoglio D., Ceci C., Garufi A., D'Orazi G. Apoptosis as anticancer mechanism: function and dysfunction of its modulators and targeted therapeutic strategies. Aging, 2016; 8(4): 603-619. Crossref ● PubMed ● PMC ● Google Scholar | ||||
| ||||
31. Wu H., Medeiros L.J., Young K.H. Apoptosis signaling and BCL-2 pathways provide opportunities for novel targeted therapeutic strategies in hematologic malignances. Blood Reviews, 2018; 32(1): 8-28. Crossref ● PubMed ● Google Scholar | ||||
| ||||
32. Yoshida H., Kong Y.Y., Yoshida R., Elia A.J., Hakem A., Hakem R., Penninger J.M., Mak T.W. Apaf1 is required for mitochondrial pathways of apoptosis and brain development. Cell, 1998; 94(6): 739-750. Crossref ● PubMed ● Google Scholar |
Refbacks
- There are currently no refbacks.
Copyright (c) 2020 Studia Biologica
This work is licensed under a Creative Commons Attribution 4.0 International License.