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GSTs, MRP and Apoptototic Markers in DLD-1 Human Colon Cancer Cell Line Before and After 5-FU Treatment

Year 2022, Volume 6, Issue 1, 25 - 40, 01.06.2022
https://doi.org/10.47947/ijnls.1073707

Abstract

Multidrugresistance is an important factor limiting the effect of chemotherapy on cancer treatment. Disorders of drug transport and apoptosis, deterioration of redox homeostasis are among the main mechanisms that lead to multidrug resistance. The aim of this study was to determine the effect of 5-FU on GST isozymes, drug resistance proteins and apoptotic proteins before and after 5-Flourouracil application on DLD-1 colon cancer cell line. The cytotoxic effect of 5-FU was measured by WST-1test and, the efficiency of drug application was, also, proved by double staining via Hoechst 33342 with Propidium iodide. Next, the expression levels of GST isozymes, drug resistance proteins and apoptotic proteins were determined by immunocytochemistry. The cytotoxic effect of 5-FU at different doses on DLD-1 colon cancer cell line was determined by WST-1 method. MRP-2, 3, 6, 7 of drug resistance proteins; GSTA1, GSTM1, GSTT1, GSTZ1, GSTK1 and GSTO1 of GST proteins; bcl-2, caspase-3, p38, and p53, which are apoptotic proteins, have higher expression in the drug-treated DLD-1 cell line. GSTS1, MDR-1 and MRP-1expressions were not immunocytochemically different. It was determined that there is a direct correlation between the level of cytotoxicity and applied drug concentration. The cytotoxic effect of the drug increased with the increase in the dose of the drug. In this study, as first in the literature, the expression levels of some apoptotic markers, GST isozymes and drug resistance proteinswere evaluated togetherand except GSTS1, MDR-1 and MRP-1, they were all upregulated with respect to the control group after 5-FU administration.

References

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Year 2022, Volume 6, Issue 1, 25 - 40, 01.06.2022
https://doi.org/10.47947/ijnls.1073707

Abstract

References

  • Ahnen, D. J., Feigl, P., Quan, G., Feneoglio-Preiser, C., Lovata, L. C., Bunn, P. A., Stemmerman, G., Wells, J. D., Macdonald, J. S., & Meyskens, Jr., F. L. (1998). Ki-ras mutation and p53 overexpression predict the clinical behavior of colorectal cancer: a Southwest Oncology Group study. Cancer Research, 58, 1149-1158.
  • Baguley, B. C. (2010). Multidrug resistance in cancer. Methods Molecular Biology 596, 1-14. https://doi.org/10.1007/978-1-60761-416-6_1
  • Bahrami, A., Amerizadeh, F., Hassanian, S., M., ShahidSales, S., Khazaei, M., Maftauh, M., Ghayour-Mobarhan, M., Ferns, G. A., & Avan, A. (2018). Genetic variants as potential predictive biomarkers in advanced colorectal cancer patients treated with oxaliplatin‐based chemotherapy. Journal of Cellular Physiology, 233(3), 2193-2201. https://doi.org/10.1002/jcp.25966
  • Cancer Today [Internet]. (2021), [cited 2021 Jun 30]. Available from: https://gco.iarc.fr/today/fact-sheets-cancers
  • Chao, C. C., Huang, Y. T., Ma, C. M., Chou, W. Y., & Lin-Chao, S. (1992). Overexpression of glutathione S-transferase and elevation of thiol pools in a multidrug-resistant human colon cancer cell line. Molecular Pharmacology, 41(1), 69-75.
  • Choi, J. H., Yoon, J. S., Won, Y. W., Park, B. B., & Lee, Y. Y. (2012). Chloroquine enhances the chemotherapeutic activity of 5-fluorouracil in a colon cancer cell line via cell cycle alteration. Acta Pathologica, Microbiologica, et Immunologica Scandinavica, 120(7), 597-604. https://doi.org/10.1111/j.1600-0463.2012.02876.x
  • Cunningham, D., Atkin, W., Lenz, H. J., Lynch, H. T., Minsky, B., Nordlinger, B., & Starling, N. (2010). Colorectal cancer. Lancet, 375, 1030-1047. https://doi.org/10.1016/S0140-6736(10)60353-4
  • Çiftçi, H., Türk, M., Tamer, U., Karahan, S., & Menemen, Y. (2013). Silver nanoparticles: cytotoxic, apoptotic, and necrotic effects on MCF-7 cells. Turkish Journal of Biology, 37, 573-581. https://doi.org/10.3906/biy-1302-21
  • Dang, D. T., Chen, F., Kohli, M., Rago, C., Cummins, J. M., & Dang, L. H. (2005). Glutathione S-transferase pi1 promotes tumorigenicity in HCT116 human colon cancer cells. Cancer Research, 65(20), 9485-9494. https://doi.org/10.1158/0008-5472.CAN-05-1930
  • Dongxing, C, Shaolan, Q., Mu, Y., & Ming, Z. (2017). The role of MRP1 in the multidrug resistance of colorectal cancer. Oncology Letters, 13, 2471-2476, https://doi.org/10.3892/ol.2017.5741
  • Elsaleh, H., Powell, B., McCaul, K, Grieu, F., Grant, R., Joseph, D., & Lacopetta, B. (2001). P53 alteration and microsatellite instability have predictive value for survival benefit from chemotherapy in stage III colorectal carcinoma. Clinical Cancer Research, 7, 1343-1349.
  • Fiore, D., Proto, M. C., Pisanti, S., Picardi, P., Pagano Zottola, A. C., Butini, S., Gemma, S., Casagni, A., Laezza, C., Vitale, M., Ligresti, A., Di Marzo, V., Zisterer, D. M., Nathwani, S., Williams, D. C., Campiani, G., Gazzerro, P., & Bifulco, M. (2016). Antitumor effect of pyrrolo-1,5-benzoxazepine-15 and its synergistic effect with Oxaliplatin and 5-FU in colorectal cancer cells. Cancer Biology & Therapy, 17(8), 849–858. https://doi.org/10.1080/15384047.2015.1078028
  • Fojo, A., Hamilton, T. C., Young, R. C., & Ozols, R. F. (1987). Multidrugresistance in ovarian cancer. Cancer, 60, 2075–80. https://doi.org/10.1002/1097-0142(19901015)60
  • Geng, M., Wang, L., & Peifeng, L. (2013). Correlation between chemosensitivity to anticancer drugs and Bcl-2 expression in gastric cancer. International Journal of Clinical and Experimental Pathology, 6(11), 2554-2559.
  • Ghafouri-Fard, S., Abak, A., TondroAnamag, F., Shoorei, H., Fattahi, F., Javadinia, S. A., Basiri, A., & Taheri, M. (2021). 5-Fluorouracil: A narrative review on the role of regulatory mechanisms in driving resistance to this chemotherapeutic agent. Frontiers in Oncology, 11, 658-636. https://doi.org/10.3389/fonc.2021.658636
  • Gong, J., Jaiswal, R., Mathys, J-M., Combes, V., Grau, G. E. R., & Bebawy, M. (2012). Microparticles and their emerging role in cancer multidrug resistance. Cancer Treatment Reviews, 38, 226–234. https://doi.org/10.1016/j.ctrv.2011.06.005
  • Gong, R. H., Yang, D. J., Kwan, H. Y., Lyu, A. P., Chen, G. Q., & Bian, Z. X. (2022). Cell death mechanisms induced by synergistic effects of halofuginone and artemisinin in colorectal cancer cells. International Journal of Medical Sciences, 19(1), 175-185. https://doi.org/10.7150/ijms.66737
  • Gottesman, M. M., & Living, V. (2006). The moleculer basis of multi drug resistanace in cancer; the early years of P-glycoprotein research. European Biochemical Societies Letter, 580, 998-1009. https://doi.org/10.1016/j.febslet.2005.12.060
  • Gottesman, M. M., Fojo, T., & Bates, S. E. (2002). Multidrugresistance in cancer: role of ATP-dependent transporters. Nature Reviews Cancer, 2, 48–58.
  • Hoban, P. R., Robson, C. N., Davies, S. M., Hall, A. G., Cattan, A. R., Hickson, I. D., & Harris, A. L. (1992). Reduced topoisomerase II and elevated alpha class glutathione S-transferase expression in a multidrug resistant CHO cell line highly cross-resistant to mitomycin C. Biochemical Pharmacology 43(4), 685-93. https://doi.org/ 10.1016/0006-2952(92)90231-7
  • Holohan, C., Van Schaeybroeck, S., Longley, D. B., & Johnston, P. G. (2013). Cancer drug resistance: an evolving paradigm. Nature Reviews Cancer, 13, 714-726. https://doi.org/ 10.1038/nrc3599
  • Jung, J. H., Taniguchi, K., Min Lee, H., Young Lee, M., Bandu, R., Komura, K., Yeon Lee, K., Akao, Y., & Pyo Kim, K. (2020). Comparative lipidomics of 5-Fluorouracil–sensitive and –resistant colorectal cancer cells reveals altered sphingomyelin and ceramide controlled by acid sphingomyelinase (SMPD1). International Journal of Scientific Reports, 10, 6124. https://doi.org/10.1038/s41598-020-62823-0
  • Khoo, E., O’Neill, S., Brown, E., Wigmore, S. J., & Harrison, E. M. (2016). Systematic review of systemic adjuvant, neoadjuvan tand perioperative chemotherapy for resectable colorectal-liver metastases. Hepato Pancreato Biliary Association, 18(6), 485-493. https://doi.org/10.1016/j.hpb.2016.03.001
  • Koçdoğan Kaya, A., Oğuztüzün, S., Şimşek, G. G., & Türk, M. (2020). Investigation of GST isoenzymes, multi drug resistance proteins, apoptotic effect in MCF-7 human breast cancer cell line before and after doxorubucin treatment. Istanbul Gelisim University Journal of Health Sciences, 10, 1-18. https://doi.org/10.38079/igusabder.631695
  • Laborde, E. (2010). Glutathione transferases as mediators of signaling pathways involved in cell proliferation and cell death. Cell Death Differ, 17, 1373–1380. https://doi.org/10.1038/cdd.2010.80
  • Liang, J. T., Huang, K. C., Cheng, Y. M, Hsu, H-Y., Cheng, A-L., Hsu, C-H., Yeh, K-H., Wang, S-H., & Chang, K-J. (2002). P53 overexpression predicts poor chemosensitivity to high-dose 5-fluorouracil plus leucovorin chemotherapy for stage IV colorectal cancers after palliative bowel resection. International Journal of Cancer, 97, 451-457. https://doi.org/10.1002/ijc.1637
  • Longley, D.B., Harkin, D. P., & Johnston, P. G. (2003). 5-fluorouracil: mechanisms of action and clinical strategies. Nature Reviews Cancer, 3, 330–338.
  • Longley, D., B., & Johnston, P. G. (2005). Molecular mechanisms of drug resistance. The Journal of Pathology, 205, 275–292. https://doi.org/10.1002/path.1706
  • Morrow, C. S., Peklak-Scott, C., Bishwokarma, B., Kute, T. E., Smitherman, P. K., & Townsend, A. J. (2006). Multidrug resistance protein 1 (MRP1, ABCC1) mediates resistance to mitoxantrone via glutathione-dependent drug efflux. Molecular Pharmacology, 69(4), 1499-505. https://doi.org/10.1124/mol.105.017988
  • Nakatsu, N., Yoshida, Y., Yamazaki, K., Nakamura, T., Dan, S., Fukui, Y., & Yamori, T. (2005). Chemosensitivity profile of cancer cell lines and identification of genes determining chemosensitivity by an integrated bioinformatical approach using cDNA arrays. Molecular Cancer Neuro Therapy, 4(3), 399-412 https://doi.org/10.1158/1535-7163.MCT-04-0234
  • Namwat, N., Amimanan, P., Loilome, W., Jearanaikoon, P., Sripa, B., Bhudhisawasdi, V., & Tassaneeyakul, W. (2008). Characterization of 5-Fluorouracil-Resistant cholangiocarcinoma cell lines. Chemotherapy, 54, 343-351. https://doi.org/10.1159/000151541
  • Neve, R. M., Chin, K., Fridlyand, J., Yeh, J., Baehner, F. L., Fevr, T., Clark, L., Bayani, N., Coppe J-P., Tong, F., Speed, T., Spellman, T. S., De Vries, S., Lapuk, A., Wang, N. J., Kuo, W-L., Stilwell, J. L., Pinkel, D., Albertson, D. G., Waldman, F. M., McCormick, F., Dickson, R. B., Johnson, M. D., Lippman, M., Etheir, S., Gazdar, A., & Gray, J. W. (2006). A collection of breast cancer cell lines for the study of functionally distinct cancer subtypes. Cancer Cell, 10(6), 515–527. https://doi.org/10.1016/j.ccr.2006.10.008
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Details

Primary Language English
Subjects Biology
Journal Section Research articles
Authors

Fatıma Nurdan GÜRBÜZ> (Primary Author)
KIRIKKALE UNIVERSITY
0000-0001-5649-954X
Türkiye


Arzu KAYA KOÇDOĞAN>
ISTANBUL GELISIM UNIVERSITY
0000-0002-3689-3061
Türkiye


Can YILMAZ>

0000-0002-0028-6614
Türkiye


Serpil OĞUZTÜZÜN>
KIRIKKALE UNIVERSITY
0000-0002-5892-3735
Türkiye


Gülçin GÜLER ŞİMŞEK>
UNIVERSITY OF HEALTH SCIENCES, GÜLHANE SCHOOL OF MEDICINE, GÜLHANE MEDICINE PR. (ANKARA)
0000-0001-7710-4631
Türkiye


Mustafa TÜRK>
KIRIKKALE UNIVERSITY
0000-0001-8202-090X
Türkiye

Supporting Institution This study was supported by Kırıkkale University Scientific Research Projects Coordination Unit
Project Number 2016/052
Thanks We thank the Scientific Research Projects Coordination Unit for their financial support to our project.
Early Pub Date January 16, 2022
Publication Date June 1, 2022
Application Date February 15, 2022
Acceptance Date April 3, 2022
Published in Issue Year 2022, Volume 6, Issue 1

Cite

APA Gürbüz, F. N. , Kaya Koçdoğan, A. , Yılmaz, C. , Oğuztüzün, S. , Güler Şimşek, G. & Türk, M. (2022). GSTs, MRP and Apoptototic Markers in DLD-1 Human Colon Cancer Cell Line Before and After 5-FU Treatment . International Journal of Nature and Life Sciences , 6 (1) , 25-40 . DOI: 10.47947/ijnls.1073707