(P12-003-25) The Effect of Vitamin E Administration on the Recovery of Colon Cancer Cells Transduced with C. elegans Fat1 Gene Post Scratch Assay

Objectives: We hypothesized that administration of vitamin E to colon cancer cell lines recovery post scratch assay would not be significantly affected when compared to cell lines not exposed to vitamin E.

Methods: Caco-2 and LoVo cell lines were transduced with C. elegans FAT1 gene. Cell lines were cultured in 24-well plates containing DMEM with and without puromycin. The cell lines were allowed to adhere to the plate surface before a scratch assay was performed. Photographs were taken at 0, 12, 24, 48, and 72hrs post scratch. Wound healing size tool was used to determine the percentage change in area of the scratch over time. evaluate the area that was scratched. The percent change was calculated for each cell line in each well and 2-way ANOVA was performed to determine the p-values.

Results: Colon cancer cell death was observed at 100ug/mL vitamin E indicating a toxic exposure level. Caco-2 cells recovered from a scratch in 3 days +/- 1 day while Caco-2 exposed to Vitamin E recovered from a scratch in 3 days +/- 1 day with a p-value range of 0.9911 at 100ug/mL of Vitamin E to a p-value of 0.7753 at 6.25ug/mL of Vitamin E administration Additionally, we found that Caco-2 and LoVo cell lines transduced with FAT1 gene recovered post scratch assay at 3 days +/- 1 day compared to Caco-2 and LoVo cell lines transduced with FAT1 exposed to Vitamin E recovered at 3 days +/- 1 day. The p-value for Caco-2 FAT1 and LoVo FAT1 transduced cells recovery time shared a p-value of >0.9999 across all Vitamin E concentrations tested, further proving the hypothesis that Vitamin E administration does not significantly effect colon cancer cell recovery post scratch assay. Using secondary data analysis, and confirmatory rt-qPCR, we observed increases in genes involved in oxidation-reduction reactions including glutathione peroxidase, superoxide dismutase, and nuclear factor erythroid 2-related factor.

Conclusions: Expression of GPX3, SOD1, and NFE2L2 antioxidant genes in colon cancer cells likely reduced the beneficial effect of vitamin E in recovery from mechanical damage. Vitamin E supplementation may not benefit patients recovering from surgical removal of colon cancer cells.

Funding Sources: Lake Erie College of Osteopathic Medicine