Squamous cell lung carcinoma is a disease in which abnormal cells within the lung divide slowly but uncontrollably, creating a tumor that can spread to other parts of the body . It is most commonly known to result from smoking or from second-hand smoke. 30% of all lung cancers are listed as squamous cell lung cancer, and can spread to the brain, adrenal glands, and liver . It is a non-small cell lung cancer (NSCLC), and that begin in the squamous cells, which are apart of the airways within the lungs . There are many treatments for lung cancer depending on the stage of the cancer, such as, chemotherapy, radiation therapy or having surgery to remove the tumors within the lungs . Lymph nodes are removed during surgery as well to check if further treatments are needed . Finding the cure to lung cancer would include completely stopping the proliferation of the cancer cells. Unfortunately, the current treatments lack the ability to do so, even though they may delay the proliferation, they do not completely stop and/or kill the cancer cells. For our experiment, we will be using NCI-H810 lung carcinoma cells. Certain enzymes can increase proliferation of cells. Histone acetyltransferase (HAT), for example, typically leads to DNA replication, that helps carry out proliferation . Within NCI-H810 cells, HATs loosen the histone proteins around the DNA nucleotides causing replication to be accessible . Garcinol, a HAT inhibitor, is gathered from the plant Garcinia Indica which has been found to be strongly effective in cancer cells, and it is able to control tumors that are in living organisms . It plays a crucial role in increasing apoptosis and stopping proliferation of cancer cells . In previous studies, Garcinol has been studied to cause apoptosis of human prostate and cervical cancer cells . There are no current studies done on garcinol and its effects on NCI-H810 cells, although in similar studies, garcinol, has shown a current status of anti-cancer effects . Our objective for this study is to determine if garcinol will cause apoptosis in the NCI-H810 cells and decrease their proliferation rate.
Hypothesis: If garcinol is applied to NCI-H810 cancer cells, then the reproduction rate of the NCI-H810 cells will decrease and many cells will undergo apoptosis causing their viability to decrease and their death rate to increase, because garcinol inhibits HAT enzymes which then do not allow replication to occur.
Methodology: We will be gathering our information over a four day period with a total of three trials. We will use the MTT assay for the proliferation assessment and the Trypan blue exclusion assay for viability and death rate assessments. We will set up three experimental groups of garcinol. Each experimental group will have 5mM, 10mM, or 20mM of garcinol.
Aim #1: To determine how garcinol affects the proliferation of NCI-H810 cells
In previous studies, garcinol has been known to stop the proliferation of cancer cells. However, there has been no studies done on garcinol and how it stops the proliferation of NCI-H810 cells. We will conduct this aim with the MTT assay to investigate how the proliferation of NCI-H810 cells is affected when exposed to garcinol. We expect that the garcinol will lower the proliferation rate of the NCI-H810 cells. We expect that the NCI-H810 cells will stop proliferating due to exposure to garcinol . This would show that garcinol has the potential to stop the spread of NCI-H810 lung cancer cells.
Aim #2: To determine the viability of NCI-H810 cells when exposed to Garcinol
In previous studies, garcinol has been known to corrupt the viability of cancer cells. However, there has been no studies done on garcinol and how it corrupts the viability of NCI-H810 cells. We will be testing with the Trypan blue exclusion assay on how garcinol corrupts the NCI-H810 cells ability to grow and divide. We expect the cells exposed to garcinol to have less functioning or living cells than those that are not exposed to garcinol . Our results would show that garcinol can have the ability to kill the NCI-H810 cells.
Aim 3: To determine the death rate of NCI-H810 cells when exposed to garcinol
In previous studies, garcinol has been known to kill cancer cells. However, there has been no studies done on garcinol and how it kills NCI-H810 cells. We will investigate with Trypan blue exclusion assay how probable garcinol is to potentially have the ability to kill the NCI-H810 cells. We expect the NCI-H810 cells to have an immense amount of dead cells where the garcinol is exposed . This would show that the NCI-H810 cells would no longer be able to replicate themselves due to garcinol inhibiting the HATs.
Broader Impact: Previous studies have reported that garcinol possesses antibiotic activities and induction of apoptosis in human leukemia HL-60 cells . Through conducting this study, without the HATs, the NCI-H810 cells will not have the ability to proliferate and they will most likely carry out apoptosis due to the loss of connection to its DNA causing the cell to fail to perform proliferation. Knowing that cells cannot proliferate without HAT inhibitors could lead to great discoveries in medicines and treatments that could lead to the cure to cancer. This is important because many people are exposed to various carcinogens, whether it be self-induced or second-hand exposure, such as smoking or pollution. Finding the cure to this cancer can help those diagnosed and potentially end the fight against all cancer.
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