An in vitro cytotoxicity of glufosfamide in HepG2 cells relative to its nonconjugated counterpart
Abstract Background Glufosfamide (β-d-glucosylisophosphoramide mustard, GLU) is an alkylating cytotoxic agent in which ifosforamide mustard (IPM) is glycosidically linked to the β-d-glucose molecule. GLU exerted its cytotoxic effect as a targeted chemotherapy. Although, its cytotoxic efficacy in a n...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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SpringerOpen
2021-08-01
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| Series: | Journal of the Egyptian National Cancer Institute |
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| Online Access: | https://doi.org/10.1186/s43046-021-00080-6 |
| _version_ | 1818910795636932608 |
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| author | Doaa E. Ahmed Fatma B. Rashidi Heba K. Abdelhakim Amr S. Mohamed Hossam M. M. Arafa |
| author_facet | Doaa E. Ahmed Fatma B. Rashidi Heba K. Abdelhakim Amr S. Mohamed Hossam M. M. Arafa |
| author_sort | Doaa E. Ahmed |
| collection | DOAJ |
| description | Abstract Background Glufosfamide (β-d-glucosylisophosphoramide mustard, GLU) is an alkylating cytotoxic agent in which ifosforamide mustard (IPM) is glycosidically linked to the β-d-glucose molecule. GLU exerted its cytotoxic effect as a targeted chemotherapy. Although, its cytotoxic efficacy in a number of cell lines, there were no experimental or clinical data available on the oncolytic effect of oxazaphosphorine drugs in hepatocellular carcinoma. Therefore, the main objective of the current study is to assess the cytotoxic potential of GLU for the first time in the hepatocellular carcinoma HepG2 cell line model. Methods Cytotoxicity was assayed by the MTT method, and half-maximal inhibitory concentration (IC50) was calculated. Flow cytometric analysis of apoptosis frequencies was measured by using Annexin V/PI double stain, an immunocytochemical assay of caspase-9, visualization of caspase-3, and Bcl2 gene expression were undertaken as apoptotic markers. Mitochondrial membrane potential was measured using the potentiometric dye; JC-1, as a clue for early apoptosis as well as ATP production, was measured by the luciferase-chemiluminescence assay. Results Glufosfamide induced cytotoxicity in HepG2 cells in a concentration- and time-dependent manner. The IC50 values for glufosfamide were significantly lower compared to ifosfamide. The frequency of apoptosis was much higher for glufosfamide than that of ifosfamide. The contents of caspase-9 and caspase-3 were elevated following exposure to GLU more than IFO. The anti-apoptotic Bcl2 gene expression, the mitochondrial membrane potential, and the cellular ATP levels were significantly decreased than in case of ifosfamide. Conclusions The current study reported for the first time cytotoxicity activity of glufosfamide in HepG2 cells in vitro. The obtained results confirmed the higher oncolytic activity of glufosfamide than its aglycone ifosfamide. The generated data warrants further elucidations by in vivo study. |
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| format | Article |
| id | doaj.art-925bd8d19f294514bfff67e3f4730b11 |
| institution | Directory Open Access Journal |
| issn | 2589-0409 |
| language | English |
| last_indexed | 2024-12-19T22:48:29Z |
| publishDate | 2021-08-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | Journal of the Egyptian National Cancer Institute |
| spelling | doaj.art-925bd8d19f294514bfff67e3f4730b112022-12-21T20:02:54ZengSpringerOpenJournal of the Egyptian National Cancer Institute2589-04092021-08-0133111510.1186/s43046-021-00080-6An in vitro cytotoxicity of glufosfamide in HepG2 cells relative to its nonconjugated counterpartDoaa E. Ahmed0Fatma B. Rashidi1Heba K. Abdelhakim2Amr S. Mohamed3Hossam M. M. Arafa4Department of Biochemistry, Misr University for Science & Technology (MUST)Biochemistry Lab.Department of Chemistry, Faculty of Science, Cairo UniversityBiochemistry Lab.Department of Chemistry, Faculty of Science, Cairo UniversityBiochemistry Lab.Department of Chemistry, Faculty of Science, Cairo UniversityDepartment of Pharmacology and Toxicology, Faculty of Pharmacy Ahram Canadian UniversityAbstract Background Glufosfamide (β-d-glucosylisophosphoramide mustard, GLU) is an alkylating cytotoxic agent in which ifosforamide mustard (IPM) is glycosidically linked to the β-d-glucose molecule. GLU exerted its cytotoxic effect as a targeted chemotherapy. Although, its cytotoxic efficacy in a number of cell lines, there were no experimental or clinical data available on the oncolytic effect of oxazaphosphorine drugs in hepatocellular carcinoma. Therefore, the main objective of the current study is to assess the cytotoxic potential of GLU for the first time in the hepatocellular carcinoma HepG2 cell line model. Methods Cytotoxicity was assayed by the MTT method, and half-maximal inhibitory concentration (IC50) was calculated. Flow cytometric analysis of apoptosis frequencies was measured by using Annexin V/PI double stain, an immunocytochemical assay of caspase-9, visualization of caspase-3, and Bcl2 gene expression were undertaken as apoptotic markers. Mitochondrial membrane potential was measured using the potentiometric dye; JC-1, as a clue for early apoptosis as well as ATP production, was measured by the luciferase-chemiluminescence assay. Results Glufosfamide induced cytotoxicity in HepG2 cells in a concentration- and time-dependent manner. The IC50 values for glufosfamide were significantly lower compared to ifosfamide. The frequency of apoptosis was much higher for glufosfamide than that of ifosfamide. The contents of caspase-9 and caspase-3 were elevated following exposure to GLU more than IFO. The anti-apoptotic Bcl2 gene expression, the mitochondrial membrane potential, and the cellular ATP levels were significantly decreased than in case of ifosfamide. Conclusions The current study reported for the first time cytotoxicity activity of glufosfamide in HepG2 cells in vitro. The obtained results confirmed the higher oncolytic activity of glufosfamide than its aglycone ifosfamide. The generated data warrants further elucidations by in vivo study.https://doi.org/10.1186/s43046-021-00080-6Glufosfamide HepG2 cytotoxicityIfofosfamide (IFO)Bcl-2Caspase-9Mitochondrial membrane potential |
| spellingShingle | Doaa E. Ahmed Fatma B. Rashidi Heba K. Abdelhakim Amr S. Mohamed Hossam M. M. Arafa An in vitro cytotoxicity of glufosfamide in HepG2 cells relative to its nonconjugated counterpart Journal of the Egyptian National Cancer Institute Glufosfamide HepG2 cytotoxicity Ifofosfamide (IFO) Bcl-2 Caspase-9 Mitochondrial membrane potential |
| title | An in vitro cytotoxicity of glufosfamide in HepG2 cells relative to its nonconjugated counterpart |
| title_full | An in vitro cytotoxicity of glufosfamide in HepG2 cells relative to its nonconjugated counterpart |
| title_fullStr | An in vitro cytotoxicity of glufosfamide in HepG2 cells relative to its nonconjugated counterpart |
| title_full_unstemmed | An in vitro cytotoxicity of glufosfamide in HepG2 cells relative to its nonconjugated counterpart |
| title_short | An in vitro cytotoxicity of glufosfamide in HepG2 cells relative to its nonconjugated counterpart |
| title_sort | in vitro cytotoxicity of glufosfamide in hepg2 cells relative to its nonconjugated counterpart |
| topic | Glufosfamide HepG2 cytotoxicity Ifofosfamide (IFO) Bcl-2 Caspase-9 Mitochondrial membrane potential |
| url | https://doi.org/10.1186/s43046-021-00080-6 |
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