Esophagus cancer (EC) is the 8th most common cancer in the world1, which makes it a high mortality rate cancer. It is estimated that about 17,000 cancer cases will be diagnosed in the USA with 18% of patients having 5-years survival rate2. Adenocarcinoma variants of EC is more common in the USA due to obesity and acid reflux problems3. Moreover, 50% of EC patient develops metastatic EC, which is hard to treat4. To date, very few drug treatments seem to be effective only against locally advanced EC, which implies the necessity of the treatment for metastatic EC5.
Coumarin is a flavoring compound naturally found in plants, which contains a benzopyrone ring. Coumarin and its derivatives (called coumarins) are found in several plants such as…show more content… Besides the vehicle control group, a CDK cell cycle inhibitor and a selective Bax activator will be used as positive controls based on previous literature. MTT assay and CyQUANT direct cell proliferation assay (Thermo Fisher Scientific) will be conducted to measure the cell viability, whereas Scratch assay and Boyden chamber assay will be conducted to measure the cell migration. The most effective coumarin derivative will be selected in terms of inhibiting the cell proliferation and migration, and potencies from in vitro study will be used to determine the dose and the concentration for in vivo study and specific aim 2 respectively. The orthotopic patient-derived xenografted mice model will be used for in vivo study. Tumor volume will be measured using magnetic resonance imaging (MRI) every week for 2 months. After that, mice will be sacrificed and organs (breast, brain, lungs, and stomach) will be tested for any possible metastasis.
Specific aim 2: To investigate the role of coumarin on EC cell cycle and apoptosis. After synchronizing the cell cycle, adenocarcinoma cell line will be treated with various concentration of the coumarin selected from aim 1. Flow cytometry will be performed to determine the stage of the cell division. Bax protein expression will be measured using the western blot method. Moreover, cell viability and cell migration assays will be repeated using a Bax inhibitor to observe the Bax induced apoptotic effect of coumarin.