Evaluation of Anti-Cancer, Pro-Apoptotic, and Anti-Metastatic Effects of Synthetic Analogue Compound 2-Imino-7-Methoxy-4-(4-fluorophenyl)-2H-1,3-Thiazino [3,2-a] Benzimidazole on Pancreatic PaCa-2 and Melanoma A375 Cancer Cells
-
Mpho Ndou
,
Marthe C. D. Fotsing
,
Michael H. K. Kengne
,
Edwin M. Mmutlane
,
Derek T. Ndinteh
,
Mthokozisi B.C. Simelane
,
Lesetja Motadi
,
Mpho Choene
Abstract
Synthetic compounds are widely used in cancer drug discovery. Chemotherapies aim to inhibit proliferation and induce apoptosis however, they have limitations. This study aims to explore in vitro anti-proliferative, pro-apoptotic, and anti-metastatic effects of 2-imino-7-methoxy-4-(4-fluorophenyl)-2h-1,3-thiazine [3,2-a] benzimidazole against pancreatic and melanoma cancer cell lines. Cell viability assays were conducted using Alamar blue assay. Caspase 3/7 activation was evaluated using caspase-Glo® 3/7 substrate reagent. Gene expression was analyzed using conventional polymerase chain reaction (PCR). Cell migration was assessed using wound healing. Alamar blue assay showed that the molecule studied exhibited antiproliferative activity on both the PaCa-2 and A375 cell lines, however, with higher cytotoxicity on PaCa-2 which suggests that it is cell-line dependent. Caspase 3/7 activity was upregulated in PaCa-2 and downregulated in A375, suggesting caspase-dependent and caspase-independent cell death, respectively. p53 and Bax were upregulated on both cell lines which suggests that the compound might have induced apoptosis and autophagy. Wound healing showed a decreased cell migration on both cell lines an important stage of metastasis. The study suggests that the study molecule can be a promising chemotherapeutic agent in the development of new anticancer drugs.
1. Arnold M, Singh D, Laversanne M, Vignat J, Vaccarella S, Meheus F, Cust AE, et al. Global burden of cutaneous melanoma in 2020 and projections to 2040. JAMA Dermatol 2022;158:495-503.
2. Ferlay J, Colombet M, Soerjomataram I, Parkin DM, Piñeros M, Znaor A, et al. Cancer statistics for the year 2020: An overview. Int J Cancer 2021;149:778-89.
3. Loeb KR, Loeb LA. Significance of multiple mutations in cancer. Carcinogenesis 2000;21:379-85.
4. Takeshima H, Ushijima T. Accumulation of genetic and epigenetic alterations in normal cells and cancer risk. NPJ Precis oncol 2019;3:7.
5. Porta M, Fabregat X, Malats N, Guarner L, Carrato A, De Miguel A, et al. Exocrine pancreatic cancer: symptoms at presentation and their relation to tumour site and stage. Clin Transl Oncol 2005;7:189-97.
6. Ilic I, Ilic M. International patterns in incidence and mortality trends of pancreatic cancer in the last three decades: a joinpoint regression analysis. World J Gastroenterol 2022;28:4698-715.
7. Martin A, Cros J, Vullierme MP, Dokmak S, Sauvanet A, Levy P, et al. Dilatation of the main pancreatic duct of unknown origin: causes and risk factors of pre-malignancy or malignancy. Surg Endosc 2023;37:3684-90.
8. Rashid S, Shaughnessy M, Tsao H. Melanoma classification and management in the era of molecular medicine. Dermatol Clin 2023;41:49-63.
9. Arnold M, Singh D, Laversanne M, Vignat J, Vaccarella S, Meheus F, Cust AE, et al. Global burden of cutaneous melanoma in 2020 and projections to 2040. JAMA Dermatol 2022;158:495-503.
10. Siegel RL, Miller KD, Wagle NS, Jemal A. Cancer statistics, 2023. Ca Cancer J Clin 2023;73:17-48.
11. Mitchell TC, Karakousis G, Schuchter L. Melanoma. InAbeloff's Clinical Oncology 2020 Jan 1 (pp. 1034-1051). Elsevier.
12. Ghobrial IM, Witzig TE, Adjei AA. Targeting apoptosis pathways in cancer therapy. CA Cancer J Clin 2005;55:178-94.
13. Fulda S, Gorman AM, Hori O, Samali A. Cellular stress responses: cell survival and cell death. Int J Cell Biol 2010:2010:214074.
14. Elmore S. Apoptosis: a review of programmed cell death. Toxicol Pathol 2007;35:495-516.
15. Borrero LJ, El-Deiry WS. Tumor suppressor p53: Biology, signaling pathways, and therapeutic targeting. Biochim Biophys Acta Rev Cancer 2021;1876:188556.
16. Kontomanolis EN, Koutras A, Syllaios A, Schizas D, Mastoraki A, Garmpis N, et al. Role of oncogenes and tumor-suppressor genes in carcinogenesis: a review. Anticancer Res 2020;40:6009-15.
17. Sarkar A, Paul A, Banerjee T, Maji A, Saha S, Bishayee A, et al. Therapeutic advancements in targeting BCL-2 family proteins by epigenetic regulators, natural, and synthetic agents in cancer. Eur J Pharmacol 2023;944:175588.
18. Mitchell TC, Karakousis G, Schuchter L. Melanoma. InAbeloff's Clinical Oncology 2020 Jan 1 (pp. 1034-1051). Elsevier.
19. Monteiro J, Fodde R. Cancer stemness and metastasis: therapeutic consequences and perspectives. European J Cancer 2010;46:1198-203.
20. Debela DT, Muzazu SG, Heraro KD, Ndalama MT, Mesele BW, Haile DC, et al. New approaches and procedures for cancer treatment: Current perspectives. SAGE Open Med 2021;9:20503121211034366.
21. Siegel R, DeSantis C, Virgo K, Stein K, Mariotto A, Smith T, et al. Cancer treatment and survivorship statistics, 2012. CA Cancer J Clin 2012;62:220-41.
22. Zhang L, Sanagapalli S, Stoita A. Challenges in diagnosis of pancreatic cancer. World J Gastroenterol 2018;24:2047-60.
23. Hajighasemi F, Tajic S. Assessment of cytotoxicity of dimethyl sulfoxide in human hematopoietic tumor cell lines. Iran J Blood Cancer 2017;9:48-53.
24. Zandi M. Cytotoxicity of taxol in combination with vincristine and vinblastine against A375 cell line. Gene Cell Tissue 2021;8:e114359.
25. Adan A, Kiraz Y, Baran Y. Cell proliferation and cytotoxicity assays. Curr Pharm Biotechnol 2016;17:1213-21.
26. Gong JX, He Y, Cui ZL, Guo YW. Synthesis, spectral characterization, and antituberculosis activity of thiazino [3, 2-A] benzimidazole derivatives. Phosphorus Sulfur Silicon Relat Elements 2016;191:1036-41.
27. Rodríguez OA, Vergara NE, Sánchez JP, Martínez MT, Sandoval ZG, Cruz A, et al. Synthesis, crystal structure, antioxidant activity and dft study of 2-aryl-2, 3-dihydro-4H-[1, 3] thiazino [3, 2-a] benzimidazol-4-One. Molecules 2014;19:8414-33.
28. Bero J, Beaufay C, Hannaert V, Hérent MF, Michels PA, Quetin-Leclercq J. Antitrypanosomal compounds from the essential oil and extracts of Keetia leucantha leaves with inhibitor activity on Trypanosoma brucei glyceraldehyde-3-phosphate dehydrogenase. Phytomedicine 2013;20:270-4.
29. Larayetan R, Ololade ZS, Ogunmola OO, Ladokun A. Phytochemical constituents, antioxidant, cytotoxicity, antimicrobial, antitrypanosomal, and antimalarial potentials of the crude extracts of Callistemon citrinus. Evid Based Complement Alternat Med 2019:2019:5410923.
30. Apontes P, Leontieva OV, Demidenko ZN, Li F, Blagosklonny MV. Exploring long-term protection of normal human fibroblasts and epithelial cells from chemotherapy in cell culture. Oncotarget 2011;2:222-33.
31. Johari SA, Sarkheil M, Veisi S. Cytotoxicity, oxidative stress, and apoptosis in human embryonic kidney (HEK293) and colon cancer (SW480) cell lines exposed to nanoscale zeolitic imidazolate framework 8 (ZIF-8). Environ Sci Pollut Res Int 2021;28:56772-81.
32. McKenzie BA, Fernandes JP, Doan MA, Schmitt LM, Branton WG, Power C. Activation of the executioner caspases-3 and-7 promotes microglial pyroptosis in models of multiple sclerosis. J Neuroinflammation 2020;17:253.
33. Yadav P, Yadav R, Jain S, Vaidya A. Caspase‐3: a primary target for natural and synthetic compounds for cancer therapy. Chem Biol Drug Des 2021;98:144-65.
34. Perfettini JL, Reed JC, Israël N, Martinou JC, Dautry-Varsat A, Ojcius DM. Role of Bcl-2 family members in caspase-independent apoptosis during Chlamydia infection. Infect Immun 2002;70:55-61.
35. Tait SW, Green DR. Caspase-independent cell death: leaving the set without the final cut. Oncogene 2008;27:6452-61.
36. Zhang YP, Li YQ, Lv YT, Wang JM. Effect of curcumin on the proliferation, apoptosis, migration, and invasion of human melanoma A375 cells. Genet Mol Res 2015;14:1056-67.
37. Güllülü Ö, Hehlgans S, Rödel C, Fokas E, Rödel F. Tumor suppressor protein p53 and inhibitor of apoptosis proteins in colorectal cancer—A promising signaling network for therapeutic interventions. Cancers (Basel) 2021;13:624.
38. Gupta S, Kass GE, Szegezdi E, Joseph B. The mitochondrial death pathway: a promising therapeutic target in diseases. J Cell Mol Med 2009;13:1004-33.
39. CC H. Clinical implications of the p53 tumor-suppressor gene. N Engl J Med 1993;329:1318-27.
40. Xu Y, Ye H. Progress in understanding the mechanisms of resistance to BCL-2 inhibitors. Exp Hematol Oncol 2022;11:31.
41. Haldar S, Negrini M, Monne M, Sabbioni S, Croce CM. Down-regulation of bcl-2 by p53 in breast cancer cells. Cancer Res 1994;54:2095-7.
42. Naseri MH, Mahdavi M, Davoodi J, Tackallou SH, Goudarzvand M, Neishabouri SH. Up regulation of Bax and down regulation of Bcl2 during 3-NC mediated apoptosis in human cancer cells. Cancer Cell Int 2015;15:55.
43. Saeki K, Yuo A, Okuma E, Yazaki Y, Susin SA, Kroemer G, et al. Bcl-2 down-regulation causes autophagy in a caspase-independent manner in human leukemic HL60 cells. Cell Death Differ 2000;7:1263-9.
44. Kögel D, Prehn JH. Caspase-independent cell death mechanisms. InMadame Curie Bioscience Database [Internet] 2013. Landes Bioscience.
45. Lee TJ, Kim EJ, Kim S, Jung EM, Park JW, Jeong SH, et al. Caspase-dependent and caspase-independent apoptosis induced by evodiamine in human leukemic U937 cells. Mol Cancer Ther 2006;5:2398-407.
46. Yee KS, Wilkinson S, James J, Ryan KM, Vousden KH. PUMA-and Bax-induced autophagy contributes to apoptosis. Cell Death Differ 2009;16:1135-45.
2. Ferlay J, Colombet M, Soerjomataram I, Parkin DM, Piñeros M, Znaor A, et al. Cancer statistics for the year 2020: An overview. Int J Cancer 2021;149:778-89.
3. Loeb KR, Loeb LA. Significance of multiple mutations in cancer. Carcinogenesis 2000;21:379-85.
4. Takeshima H, Ushijima T. Accumulation of genetic and epigenetic alterations in normal cells and cancer risk. NPJ Precis oncol 2019;3:7.
5. Porta M, Fabregat X, Malats N, Guarner L, Carrato A, De Miguel A, et al. Exocrine pancreatic cancer: symptoms at presentation and their relation to tumour site and stage. Clin Transl Oncol 2005;7:189-97.
6. Ilic I, Ilic M. International patterns in incidence and mortality trends of pancreatic cancer in the last three decades: a joinpoint regression analysis. World J Gastroenterol 2022;28:4698-715.
7. Martin A, Cros J, Vullierme MP, Dokmak S, Sauvanet A, Levy P, et al. Dilatation of the main pancreatic duct of unknown origin: causes and risk factors of pre-malignancy or malignancy. Surg Endosc 2023;37:3684-90.
8. Rashid S, Shaughnessy M, Tsao H. Melanoma classification and management in the era of molecular medicine. Dermatol Clin 2023;41:49-63.
9. Arnold M, Singh D, Laversanne M, Vignat J, Vaccarella S, Meheus F, Cust AE, et al. Global burden of cutaneous melanoma in 2020 and projections to 2040. JAMA Dermatol 2022;158:495-503.
10. Siegel RL, Miller KD, Wagle NS, Jemal A. Cancer statistics, 2023. Ca Cancer J Clin 2023;73:17-48.
11. Mitchell TC, Karakousis G, Schuchter L. Melanoma. InAbeloff's Clinical Oncology 2020 Jan 1 (pp. 1034-1051). Elsevier.
12. Ghobrial IM, Witzig TE, Adjei AA. Targeting apoptosis pathways in cancer therapy. CA Cancer J Clin 2005;55:178-94.
13. Fulda S, Gorman AM, Hori O, Samali A. Cellular stress responses: cell survival and cell death. Int J Cell Biol 2010:2010:214074.
14. Elmore S. Apoptosis: a review of programmed cell death. Toxicol Pathol 2007;35:495-516.
15. Borrero LJ, El-Deiry WS. Tumor suppressor p53: Biology, signaling pathways, and therapeutic targeting. Biochim Biophys Acta Rev Cancer 2021;1876:188556.
16. Kontomanolis EN, Koutras A, Syllaios A, Schizas D, Mastoraki A, Garmpis N, et al. Role of oncogenes and tumor-suppressor genes in carcinogenesis: a review. Anticancer Res 2020;40:6009-15.
17. Sarkar A, Paul A, Banerjee T, Maji A, Saha S, Bishayee A, et al. Therapeutic advancements in targeting BCL-2 family proteins by epigenetic regulators, natural, and synthetic agents in cancer. Eur J Pharmacol 2023;944:175588.
18. Mitchell TC, Karakousis G, Schuchter L. Melanoma. InAbeloff's Clinical Oncology 2020 Jan 1 (pp. 1034-1051). Elsevier.
19. Monteiro J, Fodde R. Cancer stemness and metastasis: therapeutic consequences and perspectives. European J Cancer 2010;46:1198-203.
20. Debela DT, Muzazu SG, Heraro KD, Ndalama MT, Mesele BW, Haile DC, et al. New approaches and procedures for cancer treatment: Current perspectives. SAGE Open Med 2021;9:20503121211034366.
21. Siegel R, DeSantis C, Virgo K, Stein K, Mariotto A, Smith T, et al. Cancer treatment and survivorship statistics, 2012. CA Cancer J Clin 2012;62:220-41.
22. Zhang L, Sanagapalli S, Stoita A. Challenges in diagnosis of pancreatic cancer. World J Gastroenterol 2018;24:2047-60.
23. Hajighasemi F, Tajic S. Assessment of cytotoxicity of dimethyl sulfoxide in human hematopoietic tumor cell lines. Iran J Blood Cancer 2017;9:48-53.
24. Zandi M. Cytotoxicity of taxol in combination with vincristine and vinblastine against A375 cell line. Gene Cell Tissue 2021;8:e114359.
25. Adan A, Kiraz Y, Baran Y. Cell proliferation and cytotoxicity assays. Curr Pharm Biotechnol 2016;17:1213-21.
26. Gong JX, He Y, Cui ZL, Guo YW. Synthesis, spectral characterization, and antituberculosis activity of thiazino [3, 2-A] benzimidazole derivatives. Phosphorus Sulfur Silicon Relat Elements 2016;191:1036-41.
27. Rodríguez OA, Vergara NE, Sánchez JP, Martínez MT, Sandoval ZG, Cruz A, et al. Synthesis, crystal structure, antioxidant activity and dft study of 2-aryl-2, 3-dihydro-4H-[1, 3] thiazino [3, 2-a] benzimidazol-4-One. Molecules 2014;19:8414-33.
28. Bero J, Beaufay C, Hannaert V, Hérent MF, Michels PA, Quetin-Leclercq J. Antitrypanosomal compounds from the essential oil and extracts of Keetia leucantha leaves with inhibitor activity on Trypanosoma brucei glyceraldehyde-3-phosphate dehydrogenase. Phytomedicine 2013;20:270-4.
29. Larayetan R, Ololade ZS, Ogunmola OO, Ladokun A. Phytochemical constituents, antioxidant, cytotoxicity, antimicrobial, antitrypanosomal, and antimalarial potentials of the crude extracts of Callistemon citrinus. Evid Based Complement Alternat Med 2019:2019:5410923.
30. Apontes P, Leontieva OV, Demidenko ZN, Li F, Blagosklonny MV. Exploring long-term protection of normal human fibroblasts and epithelial cells from chemotherapy in cell culture. Oncotarget 2011;2:222-33.
31. Johari SA, Sarkheil M, Veisi S. Cytotoxicity, oxidative stress, and apoptosis in human embryonic kidney (HEK293) and colon cancer (SW480) cell lines exposed to nanoscale zeolitic imidazolate framework 8 (ZIF-8). Environ Sci Pollut Res Int 2021;28:56772-81.
32. McKenzie BA, Fernandes JP, Doan MA, Schmitt LM, Branton WG, Power C. Activation of the executioner caspases-3 and-7 promotes microglial pyroptosis in models of multiple sclerosis. J Neuroinflammation 2020;17:253.
33. Yadav P, Yadav R, Jain S, Vaidya A. Caspase‐3: a primary target for natural and synthetic compounds for cancer therapy. Chem Biol Drug Des 2021;98:144-65.
34. Perfettini JL, Reed JC, Israël N, Martinou JC, Dautry-Varsat A, Ojcius DM. Role of Bcl-2 family members in caspase-independent apoptosis during Chlamydia infection. Infect Immun 2002;70:55-61.
35. Tait SW, Green DR. Caspase-independent cell death: leaving the set without the final cut. Oncogene 2008;27:6452-61.
36. Zhang YP, Li YQ, Lv YT, Wang JM. Effect of curcumin on the proliferation, apoptosis, migration, and invasion of human melanoma A375 cells. Genet Mol Res 2015;14:1056-67.
37. Güllülü Ö, Hehlgans S, Rödel C, Fokas E, Rödel F. Tumor suppressor protein p53 and inhibitor of apoptosis proteins in colorectal cancer—A promising signaling network for therapeutic interventions. Cancers (Basel) 2021;13:624.
38. Gupta S, Kass GE, Szegezdi E, Joseph B. The mitochondrial death pathway: a promising therapeutic target in diseases. J Cell Mol Med 2009;13:1004-33.
39. CC H. Clinical implications of the p53 tumor-suppressor gene. N Engl J Med 1993;329:1318-27.
40. Xu Y, Ye H. Progress in understanding the mechanisms of resistance to BCL-2 inhibitors. Exp Hematol Oncol 2022;11:31.
41. Haldar S, Negrini M, Monne M, Sabbioni S, Croce CM. Down-regulation of bcl-2 by p53 in breast cancer cells. Cancer Res 1994;54:2095-7.
42. Naseri MH, Mahdavi M, Davoodi J, Tackallou SH, Goudarzvand M, Neishabouri SH. Up regulation of Bax and down regulation of Bcl2 during 3-NC mediated apoptosis in human cancer cells. Cancer Cell Int 2015;15:55.
43. Saeki K, Yuo A, Okuma E, Yazaki Y, Susin SA, Kroemer G, et al. Bcl-2 down-regulation causes autophagy in a caspase-independent manner in human leukemic HL60 cells. Cell Death Differ 2000;7:1263-9.
44. Kögel D, Prehn JH. Caspase-independent cell death mechanisms. InMadame Curie Bioscience Database [Internet] 2013. Landes Bioscience.
45. Lee TJ, Kim EJ, Kim S, Jung EM, Park JW, Jeong SH, et al. Caspase-dependent and caspase-independent apoptosis induced by evodiamine in human leukemic U937 cells. Mol Cancer Ther 2006;5:2398-407.
46. Yee KS, Wilkinson S, James J, Ryan KM, Vousden KH. PUMA-and Bax-induced autophagy contributes to apoptosis. Cell Death Differ 2009;16:1135-45.
| Files | ||
| Issue | Vol 62 No 5 (2024) | |
| Section | Original Articles | |
| DOI | https://doi.org/10.18502/acta.v62i5.17777 | |
| Keywords | ||
| Cancer Synthetic compounds PaCa-2 A375 Anti-proliferation Pro-apoptosis Anti-metastatic Gene expression p53 Bcl-2 Bax | ||
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How to Cite
1.
Ndou M, Fotsing M, Kengne M, M. Mmutlane E, T. Ndinteh D, B.C. Simelane M, Motadi L, Choene M. Evaluation of Anti-Cancer, Pro-Apoptotic, and Anti-Metastatic Effects of Synthetic Analogue Compound 2-Imino-7-Methoxy-4-(4-fluorophenyl)-2H-1,3-Thiazino [3,2-a] Benzimidazole on Pancreatic PaCa-2 and Melanoma A375 Cancer Cells. Acta Med Iran. 2025;62(5):277-289.
