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The Apoptotic Effects of a Proteasome Inhibitor on Caco-2 Colon Carcinoma Cells

Year 2023, Volume: 2 Issue: 1, 19 - 24, 18.04.2023

Abstract

The first and reversible inhibitor of the 26S proteasome (Velcade, formerly PS-341) has a crucial role with its antitumor activity against a number of malignancies and acts as a prominent inducer of apoptosis. In the present work, it was demonstrated the enhanced anticancer activity of a proteasome inhibitor by evaluating the levels of Bcl-2 protein family members on Caco-2 cell line. For this reason, Caco-2 colon carcinoma cells were treated with Velcade at four different doses (0, 5, 10, and 20nM). Western blotting was performed to detect the expression levels of apoptotic protein. Additionally, apoptotic cell death was demonstrated with in situ TUNEL staining by immunohistochemistry (IHC). An increase the expression levels of both pro-apoptotic proteins was detected in a dose-dependent manner; however, the effective dose was detected as 10 nM. According to the IHC results, it can be said that a serious increase in the amount of apoptotic cells was determined. Taking everything into account, this proteasome inhibitor has a potential to be effective on inducing apoptotic mechanism on colon carcinoma cells.

References

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  • 2. Aldahhan R, Almohazey D, Khan FA. Emerging trends in the application of gold nanoformulations in colon cancer diagnosis and treatment. Semin Cancer Biol. 2022;86(Pt 2):1056-65.
  • 3. Hong YS, Hong SW, Kim SM, Jin DH, Shin JS, Yoon DH, et al. Bortezomib induces G2-M arrest in human colon cancer cells through ROS-inducible phosphorylation of ATM-CHK1. International journal of oncology. 2012;41(1):76-82.
  • 4. Cunningham D, Humblet Y, Siena S, Khayat D, Bleiberg H, Santoro A, et al. Cetuximab monotherapy and cetuximab plus irinotecan in irinotecan-refractory metastatic colorectal cancer. The New England journal of medicine. 2004;351(4):337-45.
  • 5. Douillard JY, Cunningham D, Roth AD, Navarro M, James RD, Karasek P, et al. Irinotecan combined with fluorouracil compared with fluorouracil alone as first-line treatment for metastatic colorectal cancer: a multicentre randomised trial. Lancet. 2000;355(9209):1041-7.
  • 6. Hochster HS, Hart LL, Ramanathan RK, Childs BH, Hainsworth JD, Cohn AL, et al. Safety and efficacy of oxaliplatin and fluoropyrimidine regimens with or without bevacizumab as first-line treatment of metastatic colorectal cancer: results of the TREE Study. Journal of clinical oncology : official journal of the American Society of Clinical Oncology. 2008;26(21):3523-9.
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  • 8. Van Cutsem E, Kohne CH, Hitre E, Zaluski J, Chang Chien CR, Makhson A, et al. Cetuximab and chemotherapy as initial treatment for metastatic colorectal cancer. The New England journal of medicine. 2009;360(14):1408-17.
  • 9. de Gramont A, Figer A, Seymour M, Homerin M, Hmissi A, Cassidy J, et al. Leucovorin and fluorouracil with or without oxaliplatin as first-line treatment in advanced colorectal cancer. Journal of clinical oncology : official journal of the American Society of Clinical Oncology. 2000;18(16):2938-47.
  • 10. Pitts TM, Morrow M, Kaufman SA, Tentler JJ, Eckhardt SG. Vorinostat and bortezomib exert synergistic antiproliferative and proapoptotic effects in colon cancer cell models. Molecular cancer therapeutics. 2009;8(2):342-9.
  • 11. Fennell DA, Chacko A, Mutti L. BCL-2 family regulation by the 20S proteasome inhibitor bortezomib. Oncogene. 2008;27(9):1189-97.
  • 12. Garrido C, Brunet M, Didelot C, Zermati Y, Schmitt E, Kroemer G. Heat shock proteins 27 and 70: anti-apoptotic proteins with tumorigenic properties. Cell cycle. 2006;5(22):2592-601.
  • 13. Song X, Kim SY, Zhang L, Tang D, Bartlett DL, Kwon YT, et al. Role of AMP-activated protein kinase in cross-talk between apoptosis and autophagy in human colon cancer. Cell death & disease. 2014;5:e1504.
  • 14. Zhu K, Dunner K, Jr., McConkey DJ. Proteasome inhibitors activate autophagy as a cytoprotective response in human prostate cancer cells. Oncogene. 2010;29(3):451-62.
  • 15. Ebert B, Kisiela M, Wsol V, Maser E. Proteasome inhibitors MG-132 and bortezomib induce AKR1C1, AKR1C3, AKR1B1, and AKR1B10 in human colon cancer cell lines SW-480 and HT-29. Chem Biol Interact. 2011;191(1-3):239-49.
  • 16. Chang HK, Shin MS, Yang HY, Lee JW, Kim YS, Lee MH, et al. Amygdalin induces apoptosis through regulation of Bax and Bcl-2 expressions in human DU145 and LNCaP prostate cancer cells. Biol Pharm Bull. 2006;29(8):1597-602.
  • 17. Danial NN, Korsmeyer SJ. Cell death: critical control points. Cell. 2004;116(2):205-19.
  • 18. Yu J, Tiwari S, Steiner P, Zhang L. Differential apoptotic response to the proteasome inhibitor Bortezomib [VELCADE, PS-341] in Bax-deficient and p21-deficient colon cancer cells. Cancer biology & therapy. 2003;2(6):694-9.
  • 19. Ding WX, Ni HM, Chen X, Yu J, Zhang L, Yin XM. A coordinated action of Bax, PUMA, and p53 promotes MG132-induced mitochondria activation and apoptosis in colon cancer cells. Molecular cancer therapeutics. 2007;6(3):1062-9.
  • 20. Karin M, Yamamoto Y, Wang QM. The IKK NF-kappa B system: a treasure trove for drug development. Nature reviews Drug discovery. 2004;3(1):17-26.
  • 21. Hideshima T, Chauhan D, Richardson P, Mitsiades C, Mitsiades N, Hayashi T, et al. NF-kappa B as a therapeutic target in multiple myeloma. The Journal of biological chemistry. 2002;277(19):16639-47.
  • 22. Green DR, Kroemer G. Pharmacological manipulation of cell death: clinical applications in sight? The Journal of clinical investigation. 2005;115(10):2610-7.
  • 23. Amaravadi RK, Lippincott-Schwartz J, Yin XM, Weiss WA, Takebe N, Timmer W, et al. Principles and current strategies for targeting autophagy for cancer treatment. Clinical cancer research : an official journal of the American Association for Cancer Research. 2011;17(4):654-66.
  • 24. Waters JP, Pober JS, Bradley JR. Tumour necrosis factor and cancer. The Journal of pathology. 2013;230(3):241-8.
  • 25. Bai J, Demirjian A, Sui J, Marasco W, Callery MP. Histone deacetylase inhibitor trichostatin A and proteasome inhibitor PS-341 synergistically induce apoptosis in pancreatic cancer cells. Biochemical and biophysical research communications. 2006;348(4):1245-53.
  • 26. Jiang X, Tsang YH, Yu Q. c-Myc overexpression sensitizes Bim-mediated Bax activation for apoptosis induced by histone deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA) through regulating Bcl-2/Bcl-xL expression. The international journal of biochemistry & cell biology. 2007;39(5):1016-25.
  • 27. Pei XY, Dai Y, Grant S. Synergistic induction of oxidative injury and apoptosis in human multiple myeloma cells by the proteasome inhibitor bortezomib and histone deacetylase inhibitors. Clinical cancer research : an official journal of the American Association for Cancer Research. 2004;10(11):3839-52.
  • 28. Kretowski R, Stypulkowska A, Cechowska-Pasko M. Efficient apoptosis and necrosis induction by proteasome inhibitor: bortezomib in the DLD-1 human colon cancer cell line. Molecular and cellular biochemistry. 2015;398(1-2):165-73.
  • 29. Ciechanover A, Orian A, Schwartz AL. The ubiquitin-mediated proteolytic pathway: mode of action and clinical implications. Journal of cellular biochemistry Supplement. 2000;34:40-51.
  • 30. Bonfili L, Cuccioloni M, Cecarini V, Mozzicafreddo M, Palermo FA, Cocci P, et al. Ghrelin induces apoptosis in colon adenocarcinoma cells via proteasome inhibition and autophagy induction. Apoptosis : an international journal on programmed cell death. 2013;18(10):1188-200.
  • 31. McCray BA, Taylor JP. The role of autophagy in age-related neurodegeneration. Neuro-Signals. 2008;16(1):75-84.
Year 2023, Volume: 2 Issue: 1, 19 - 24, 18.04.2023

Abstract

References

  • 1. Kim SY, Song X, Zhang L, Bartlett DL, Lee YJ. Role of Bcl-xL/Beclin-1 in interplay between apoptosis and autophagy in oxaliplatin and bortezomib-induced cell death. Biochemical pharmacology. 2014;88(2):178-88.
  • 2. Aldahhan R, Almohazey D, Khan FA. Emerging trends in the application of gold nanoformulations in colon cancer diagnosis and treatment. Semin Cancer Biol. 2022;86(Pt 2):1056-65.
  • 3. Hong YS, Hong SW, Kim SM, Jin DH, Shin JS, Yoon DH, et al. Bortezomib induces G2-M arrest in human colon cancer cells through ROS-inducible phosphorylation of ATM-CHK1. International journal of oncology. 2012;41(1):76-82.
  • 4. Cunningham D, Humblet Y, Siena S, Khayat D, Bleiberg H, Santoro A, et al. Cetuximab monotherapy and cetuximab plus irinotecan in irinotecan-refractory metastatic colorectal cancer. The New England journal of medicine. 2004;351(4):337-45.
  • 5. Douillard JY, Cunningham D, Roth AD, Navarro M, James RD, Karasek P, et al. Irinotecan combined with fluorouracil compared with fluorouracil alone as first-line treatment for metastatic colorectal cancer: a multicentre randomised trial. Lancet. 2000;355(9209):1041-7.
  • 6. Hochster HS, Hart LL, Ramanathan RK, Childs BH, Hainsworth JD, Cohn AL, et al. Safety and efficacy of oxaliplatin and fluoropyrimidine regimens with or without bevacizumab as first-line treatment of metastatic colorectal cancer: results of the TREE Study. Journal of clinical oncology : official journal of the American Society of Clinical Oncology. 2008;26(21):3523-9.
  • 7. Bokemeyer C, Bondarenko I, Makhson A, Hartmann JT, Aparicio J, de Braud F, et al. Fluorouracil, leucovorin, and oxaliplatin with and without cetuximab in the first-line treatment of metastatic colorectal cancer. Journal of clinical oncology : official journal of the American Society of Clinical Oncology. 2009;27(5):663-71.
  • 8. Van Cutsem E, Kohne CH, Hitre E, Zaluski J, Chang Chien CR, Makhson A, et al. Cetuximab and chemotherapy as initial treatment for metastatic colorectal cancer. The New England journal of medicine. 2009;360(14):1408-17.
  • 9. de Gramont A, Figer A, Seymour M, Homerin M, Hmissi A, Cassidy J, et al. Leucovorin and fluorouracil with or without oxaliplatin as first-line treatment in advanced colorectal cancer. Journal of clinical oncology : official journal of the American Society of Clinical Oncology. 2000;18(16):2938-47.
  • 10. Pitts TM, Morrow M, Kaufman SA, Tentler JJ, Eckhardt SG. Vorinostat and bortezomib exert synergistic antiproliferative and proapoptotic effects in colon cancer cell models. Molecular cancer therapeutics. 2009;8(2):342-9.
  • 11. Fennell DA, Chacko A, Mutti L. BCL-2 family regulation by the 20S proteasome inhibitor bortezomib. Oncogene. 2008;27(9):1189-97.
  • 12. Garrido C, Brunet M, Didelot C, Zermati Y, Schmitt E, Kroemer G. Heat shock proteins 27 and 70: anti-apoptotic proteins with tumorigenic properties. Cell cycle. 2006;5(22):2592-601.
  • 13. Song X, Kim SY, Zhang L, Tang D, Bartlett DL, Kwon YT, et al. Role of AMP-activated protein kinase in cross-talk between apoptosis and autophagy in human colon cancer. Cell death & disease. 2014;5:e1504.
  • 14. Zhu K, Dunner K, Jr., McConkey DJ. Proteasome inhibitors activate autophagy as a cytoprotective response in human prostate cancer cells. Oncogene. 2010;29(3):451-62.
  • 15. Ebert B, Kisiela M, Wsol V, Maser E. Proteasome inhibitors MG-132 and bortezomib induce AKR1C1, AKR1C3, AKR1B1, and AKR1B10 in human colon cancer cell lines SW-480 and HT-29. Chem Biol Interact. 2011;191(1-3):239-49.
  • 16. Chang HK, Shin MS, Yang HY, Lee JW, Kim YS, Lee MH, et al. Amygdalin induces apoptosis through regulation of Bax and Bcl-2 expressions in human DU145 and LNCaP prostate cancer cells. Biol Pharm Bull. 2006;29(8):1597-602.
  • 17. Danial NN, Korsmeyer SJ. Cell death: critical control points. Cell. 2004;116(2):205-19.
  • 18. Yu J, Tiwari S, Steiner P, Zhang L. Differential apoptotic response to the proteasome inhibitor Bortezomib [VELCADE, PS-341] in Bax-deficient and p21-deficient colon cancer cells. Cancer biology & therapy. 2003;2(6):694-9.
  • 19. Ding WX, Ni HM, Chen X, Yu J, Zhang L, Yin XM. A coordinated action of Bax, PUMA, and p53 promotes MG132-induced mitochondria activation and apoptosis in colon cancer cells. Molecular cancer therapeutics. 2007;6(3):1062-9.
  • 20. Karin M, Yamamoto Y, Wang QM. The IKK NF-kappa B system: a treasure trove for drug development. Nature reviews Drug discovery. 2004;3(1):17-26.
  • 21. Hideshima T, Chauhan D, Richardson P, Mitsiades C, Mitsiades N, Hayashi T, et al. NF-kappa B as a therapeutic target in multiple myeloma. The Journal of biological chemistry. 2002;277(19):16639-47.
  • 22. Green DR, Kroemer G. Pharmacological manipulation of cell death: clinical applications in sight? The Journal of clinical investigation. 2005;115(10):2610-7.
  • 23. Amaravadi RK, Lippincott-Schwartz J, Yin XM, Weiss WA, Takebe N, Timmer W, et al. Principles and current strategies for targeting autophagy for cancer treatment. Clinical cancer research : an official journal of the American Association for Cancer Research. 2011;17(4):654-66.
  • 24. Waters JP, Pober JS, Bradley JR. Tumour necrosis factor and cancer. The Journal of pathology. 2013;230(3):241-8.
  • 25. Bai J, Demirjian A, Sui J, Marasco W, Callery MP. Histone deacetylase inhibitor trichostatin A and proteasome inhibitor PS-341 synergistically induce apoptosis in pancreatic cancer cells. Biochemical and biophysical research communications. 2006;348(4):1245-53.
  • 26. Jiang X, Tsang YH, Yu Q. c-Myc overexpression sensitizes Bim-mediated Bax activation for apoptosis induced by histone deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA) through regulating Bcl-2/Bcl-xL expression. The international journal of biochemistry & cell biology. 2007;39(5):1016-25.
  • 27. Pei XY, Dai Y, Grant S. Synergistic induction of oxidative injury and apoptosis in human multiple myeloma cells by the proteasome inhibitor bortezomib and histone deacetylase inhibitors. Clinical cancer research : an official journal of the American Association for Cancer Research. 2004;10(11):3839-52.
  • 28. Kretowski R, Stypulkowska A, Cechowska-Pasko M. Efficient apoptosis and necrosis induction by proteasome inhibitor: bortezomib in the DLD-1 human colon cancer cell line. Molecular and cellular biochemistry. 2015;398(1-2):165-73.
  • 29. Ciechanover A, Orian A, Schwartz AL. The ubiquitin-mediated proteolytic pathway: mode of action and clinical implications. Journal of cellular biochemistry Supplement. 2000;34:40-51.
  • 30. Bonfili L, Cuccioloni M, Cecarini V, Mozzicafreddo M, Palermo FA, Cocci P, et al. Ghrelin induces apoptosis in colon adenocarcinoma cells via proteasome inhibition and autophagy induction. Apoptosis : an international journal on programmed cell death. 2013;18(10):1188-200.
  • 31. McCray BA, Taylor JP. The role of autophagy in age-related neurodegeneration. Neuro-Signals. 2008;16(1):75-84.
There are 31 citations in total.

Details

Primary Language English
Subjects Clinical Sciences (Other)
Journal Section Research Articles
Authors

Selina Aksak Karameşe

Early Pub Date March 30, 2023
Publication Date April 18, 2023
Published in Issue Year 2023 Volume: 2 Issue: 1

Cite

APA Aksak Karameşe, S. (2023). The Apoptotic Effects of a Proteasome Inhibitor on Caco-2 Colon Carcinoma Cells. Eurasian Journal of Molecular and Biochemical Sciences, 2(1), 19-24.