The Best Fruit Extract for the Inhibition of Lung Carcinomas
University Transition Program
Floor Location : M 056 H
The purpose of my project was to determine the best inhibitor of lung carcinomas CRL-5928 and HTB-182, based on cell viability tests and cytotoxicity. Both of these cancers are categorized as bronchial squamous cell carcinomas. I cultured these cells in petri dishes, counted them, and then plated in five 96-well plates with an initial density of 1000cell/well. I tested the inhibitory effects of 3 different fruit extracts on the tumour cells at three different concentrations each: 25ug of extract/mL of media (cell food), 50ug/mL, and 100ug/mL over a period of five days. Commercially acquired grape seed, cranberry and pomegranate extracts were tested. Four replicates were performed for each well in the experiment to ensure accuracy in the data. MTT cell viability assays were conducted each day to see how many cells were still alive in the fruit extract solution. The values from these variables were compared to the values of the control, which were simply the cells plated with media. rn The experiment was repeated with normal bronchial squamous cells: HBEC. These tests would indicate how cytotoxic the fruit extract was. The data retrieved from these tests were critical as an extract that inhibits cancer cells well may also kill normal cells, which is unfavoured in chemotherapy. As the HBECs were much more vulnerable to apoptosis than tumour cells, the initial plating density of HBEC was 1500cells/well. When analyzing the data, I will organize the cell viability data for the tumour cells were divided by the cell viability of normal cells to determine which extract was less cytotoxic, but still displayed good inhibitory effects on tumour cells.rn The cells were cultured with media in petri dishes, then left for a few days to split and grow, and were then counted through a complicated process involving many pipettes (and tips), trypsin to release the cells from the dish, media to grow the cells in and deactivate the trypsin, a hemocytometer, a cell-counter and a microscope. All parts of the experimentation for this project were performed in a biological fume hood, and proper cleaning/disposal procedures followed the laboratory experiments.rn After counting the cells, I calculated how much of the cell solution I would require to mix with media in order to achieve the specified fruit extract concentration. Then, I plated the cells in 96-well plates, and left the plates in a 37 degrees celcius incubator and allowed to grow for a day before adding fruit extracts in the wells. rnrnThe Grape Seed Extract was shown to be the most effective in inhibiting cancer while keeping normal cells alive, while the cranberry extract appeared to be the least effective in inhibiting tumour cells. The pomegranate extract functioned well in inhibiting tumour cells, but also killed a significant number of normal cells.rnrnThe rest of the Results, including the quantitative and qualitative data acquired from this experiment will be displayed at the project board.