Induction of Apoptosis by Selected Natural Products in v-Cbl Transformed COS and NIH 3T3 Cells

Abstract

3 Abstract: A comparative study was made on the relative sensitivity of normal cells vis-a-vis transformed cells to the induction of apoptosis by the selected natural products. Oncoproteins predispose cells for apoptosis. Flourensia cernua, Ambrosia chamissonis and camptothecin, was used as stimuli to induce apoptosis in transformed cells. V-Cbl plasmid -whole plasmid carring the v-Cbl gene-( pGEX-KG-v-Cbl 8,000 kb) and empty vector ( pGEX-KG 5,000kb) was used for transfection of COS and NIH 3T3 cells. We hypothesized that the Cbl proteins could predispose the transformed cells to apoptosis, by making cells vulnerable to outside insult, could lead to their apoptotic death. Even with higher doses, such as 30 and 40 µL, of Flourensia cernua, apoptotic influence was not statistically significant -0.035% of all cells. Twenty microliter of Ambrosia chamissonis also produced a statistically insignificant number -0.040% of all cells. The presence of intact nuclei stained pink by Propidium Iodide was generally observed, indicating necrosis. After longer duration (72 h), a higher number of necrotic cells was always observed. Murine fibroblast NIH 3T3 cells were more often found to be necrotic than COS fibroblasts. Delivering DNA by transfection resulted in toxicity, leading to the constant presence of necrosis. Key wards: Apoptosis, fibroblast, natural products, necrosis, plasmid