1.Anti-proliferative Effects of Androctonus amoreuxi Scorpion and Cerastes cerastes Snake Venoms on Human Prostate Cancer Cells.
Hassan AKEF ; Nahla KOTB ; Dina ABO-ELMATTY ; Sayed SALEM
Journal of Cancer Prevention 2017;22(1):40-46
The present study evaluated the effects of Androctonus amoreuxi scorpion venom, Cerastes cerastes snake venom and their mixture on prostate cancer cells (PC3). An MTT assay was used to determine the anti-proliferative effect of the venoms, while quantitative real time PCR was used to evaluate the expression of apoptosis-related genes (Bax and Bcl-2). Furthermore, colorimetric assays were used to measure the levels of malondialdehyde (MDA) and antioxidant enzymes. Our results show that the venoms significantly reduced PC3 cell viability in a dose-dependent manner. On the other hand, these venoms significantly decreased Bcl-2 gene expression. Additionally, C. cerastes venom significantly reduced Bax gene expression, while A. amoreuxi venom and a mixture of A. amoreuxi & C. cerastes venoms did not alter Bax expression. Consequently, these venoms significantly increased the Bax/Bcl-2 ratio and the oxidative stress biomarker MDA. Furthermore, these venoms also increased the activity levels of the antioxidant enzymes, catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase, and glutathione-S-transferase. Overall, the venoms have cytotoxic and anti-proliferative effects on PC3 cells.
Apoptosis
;
Catalase
;
Cell Survival
;
Gene Expression
;
Genes, bcl-2
;
Glutathione Peroxidase
;
Glutathione Reductase
;
Hand
;
Humans*
;
Malondialdehyde
;
Oxidative Stress
;
Prostate*
;
Prostatic Neoplasms*
;
Real-Time Polymerase Chain Reaction
;
Scorpion Venoms
;
Scorpions*
;
Snake Venoms*
;
Snakes*
;
Superoxide Dismutase
;
Venoms
;
Viper Venoms
;
Viperidae*
2.Human platelet lysate efficiency, stability, and optimal heparin concentration required in culture of mammalian cells
Hoda E MOHAMED ; Mervat E ASKER ; Nahla S KOTB ; Akram M EL HABAB
Blood Research 2020;55(1):35-43
BACKGROUND:
Fetal bovine serum (FBS) has been used to support the growth and proliferation of mammalian cells for decades. Owing to several risk factors associated with FBS, several trials have been conducted to evaluate substitutes to FBS with the same efficiency and the lower risk issues.
METHODS:
In this study, human platelet lysate (HPL) derived from activated human platelets was evaluated as an alternative to FBS due to the associated risk factors. To evaluate the efficiency of the preparation process, platelet count was performed before and after activation. The concentrations of several growth factors and proteins were measured to investigate HPL efficiency. HPL stability was studied at regular intervals, and optimal heparin concentration required to prevent gel formation in various media was determined. The biological activity of HPL and FBS was compared by evaluating the growth performance of Vero and Hep-2 cell lines.
RESULTS:
Result of platelet count assay revealed the efficiency of HPL preparation process. Growth factor concentrations in HPL were significantly higher than those in FBS, while the protein content of HPL was lower than that of FBS. Stability study data showed that the prepared HPL was stable for up to 15 months at −20℃. Ideal heparin concentration to be used in different media was dependent on calcium concentration. Results of cell viability assay showed that HPL was superior to FBS in supporting the growth and proliferation of Vero and Hep-2 cells.
CONCLUSION
The HPL prepared by the mechanical activation of platelets may serve as an efficient alternative to FBS in cell culture process.