title: The chemomodulatory effects of glufosfamide on docetaxel cytotoxicity in prostate cancer cells.
abstract: Background. Glufosfamide (GLU) is a glucose conjugate of ifosfamide in which isophosphoramide mustard is glycosidically linked to the β-D-glucose molecule. Based on GLU structure, it is considered a targeted chemotherapy with fewer side effects. The main objective of the current study is to assess the cytotoxic potential of GLU for the first time in prostate cancer (PC) cells representing different stages of the tumor. Furthermore, this study examined the potential synergistic activity of GLU in combination with docetaxel (DOC). Methods. Two different cell lines were used, LNCaP and PC-3. Concentration-response curves were assessed. The tested groups per cell line were, control, GLU, DOC and combination. Treatment duration was 72 h. Cytotoxicity was assessed using sulforhodamine B (SRB) assay and half maximal inhibitory concentration (IC50) was calculated. Synergy analyses were performed using Calcusyn(®)software. Subsequent mechanistic studies included β-glucosidase activity assay, glucose uptake and apoptosis studies, namely annexin V-FITC assay and the protein expression of mitochondrial pathway signals including Bcl-2, Bax, Caspase 9 and 3 were assessed. Data are presented as mean ± SD; comparisons were carried out using one way analysis of variance (ANOVA) followed by Tukey-Kramer's test for post hoc analysis. Results. GLU induced cytotoxicity in both cell lines in a concentration-dependent manner. The IC50 in PC-3 cells was significantly lower by 19% when compared to that of LNCaP cells. The IC50 of combining both drugs showed comparable effect to DOC in PC-3 but was tremendously lowered by 49% compared to the same group in LNCaP cell line. β-glucosidase activity was higher in LNCaP by about 67% compared to that determined in PC-3 cells while the glucose uptake in PC-3 cells was almost 2 folds that found in LNCaP cells. These results were directly correlated to the efficacy of GLU in each cell line. Treatment of PC cells with GLU as single agent or in combination with DOC induced significantly higher apoptosis as evidenced by Annexin V-staining. Apoptosis was significantly increased in combination group by 4.9 folds and by 2.1 Folds when compared to control in LNCaP cells and PC-3 cells; respectively. Similarly, the expression of Bcl-2 was significantly decreased while Bax, caspase 9 and 3 were significantly increased in the combined treatment groups compared to the control. Conclusion. GLU has a synergistic effect in combination with DOC as it increases the cell kill which can be attributed at least partially to apoptosis in both the tested cell lines and it is suggested as a new combination regimen to be considered in the treatment of the prostate cancer. Further experiments and clinical investigations are needed for assessment of that regimen.
authors.affiliation: Department of Pharmacology, Toxicology and Biochemistry, Faculty of Pharmacy, Future University in Egypt (FUE) , Cairo , Egypt.
authors.forename: Reem T
authors.initials: RT
authors.lastname: Attia
authors.affiliation: Biology Department, The School of Sciences and Engineering, The American University in Cairo, New Cairo, Egypt; Department of Pharmacology and Toxicology, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt.
authors.forename: Mai F
authors.initials: MF
authors.lastname: Tolba
authors.affiliation: Pharmaceutical Sciences, University of Colorado Anschutz Medical Center , Aurora , CO , United States.
authors.forename: Ruchit
authors.initials: R
authors.lastname: Trivedi
authors.affiliation: Department of Pharmacology and Toxicology, Faculty of Pharmacy, Ain Shams University , Cairo , Egypt.
authors.forename: Mariane G
authors.initials: MG
authors.lastname: Tadros
authors.affiliation: Department of Pharmacology and Toxicology, Faculty of Pharmacy, Modern University for Technology and Information , Cairo , Egypt.
authors.forename: Hossam M M
authors.initials: HM
authors.lastname: Arafa
authors.affiliation: Department of Pharmacology and Toxicology, Faculty of Pharmacy, Ain Shams University , Cairo , Egypt.
authors.forename: Ashraf B
authors.initials: AB
authors.lastname: Abdel-Naim
keywords: Apoptosis; Docetaxel; Glufosfamide; LNCaP; PC-3; Prostate cancer