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The Effect of Compound L19 on Human Colorectal Cells (DLD‐1)

Colorectal cancer (CRC) is the third most common type of cancer in the world and the second leading cause of human deaths due to cancer (Siegel, et al. 2014). Rumex crispus extracts, a source of anticancer compounds, have demonstrated the induction of apoptosis in colorectal cancer cells (DLD‐1). Bhandari (2015) screened water soluble compounds from roots and leaves identifying those that induced apoptosis in DLD‐1 cells. L19 is a compound that will be studied in more detail in this project. In this research, specific genes involved in apoptosis induction as a result of treatment of the DLD‐1 cells with L19 are identified using methods already utilized by Bhandari (2015) with addition of qRT‐PCR analysis. Accelerated solvent extraction (ASE) followed by high performance liquid chromatography (HPLC) was utilized to obtain larger amounts of L19. GC‐Mass spectrometry was used to identify the isolated compound. Apoptosis was measured by detecting the levels of caspase 3 and 7 using the APO‐one assay (Promega, Corp.) after treating the DLD‐1 cells with varying concentrations of L19. Furthermore, the mechanism of apoptosis has been determined using qRT‐PCR and gene specific primers for caspase 8,10 (extrinsic pathway), 9 (instrinsic), 4, 5 and 12 (ER stress).