Discipline: Biological Sciences
Subcategory: Cancer Research
Room: Exhibit Hall A
Faren Renee White - Tougaloo College
Co-Author(s): J’Mone McClenty, Alcorn State University, Lorman, MS; Tanisha Hinton, Jackson State University, Jackson, MS; Grace Ikenga, Jackson State University, Jackson, MS; Shaloam Dasari,Jackson State University, Jackson, MS; and Clement G. Yedjou, Jackson State University, Jackson, MS
Garlic supplementation in diet has been shown to be beneficial to cancer patients. Recently, its pharmacological role in the prevention and treatment of cancer has received increasing attention. However, the mechanisms by which garlic extract induces cytotoxic effects in cancer cells remain largely unknown. The present study was designed to use HL-60 cells as a test model to determine whether garlic treatment induced toxicity to human leukemia cells is mediated through oxidative stress. Human leukemia (HL-60) cells were treated with different concentrations of garlic extract for 24 hr. Live and dead cells was determined by trypan blue exclusion test and microscopic imaging. The role of oxidative stress in garlic toxicity was assessed by lipid peroxidation, glutathione peroxidase (GPx) and catalase (Cat) assays, respectively. Oxidative stress biomarkers showed significant increase (p <0.05) of malondialdehyde levels on one hand and gradual decrease of antioxidant enzyme activity (GPx & Cat) on the other hand with increasing garlic doses. Taken together, finding from the present study demonstrates that at therapeutic concentrations, garlic treatment induced cytotoxic effects through oxidative in HL-60 cells. Although published studies indicate that garlic has medicinal properties effective against many diseases other than APL, the cellular mechanisms under which this compound exerts its therapeutic effect in cancer cells remain largely unknown. We hypothesized that treatment of HL-60 cells with garlic would lead to membrane damage and loss of viability of these cells. Therefore, the goal of the present study was to assess the therapeutic efficacy of garlic in the management of acute promyelocytic leukemia. Human promyelocytic leukemia (HL-60) cells were grown in 6-well plates in RPMI 1640 supplemented with 10% FBS, and 1% penicillin-streptomycin until about 85% full. Cells were treated at different concentrations of garlic extract (0- 10 mg/mL), and incubated at 37oC, 5% CO2 for 24 hours. Following exposure to garlic extract, the MTT assay was performed for cell viability, and cell proliferation using a microplate reader with the wavelength set at 550 nm. Briefly, cells were washed two times with PBS and then suspended in 1ml of medium at 1 x 106 cells per mL. 100 microliters of this solution was transferred to 1 ml culture tubes. 10 microliters of the dye were added to 100 uL of cells suspension. Samples were analyzed after 2 minutes by cellometer vision.Data obtained from the trypan blue exclusion test indicated that GE significantly (p < 0.05) reduced the viability of HL-60 cells in a concentration-dependent manner. Similar trend was observed in the data obtained from the MTT results.Finding from the present study demonstrates that at therapeutic concentrations, garlic treatment induced cytotoxic effects in HL-60 cells associated with oxidative stress.
Funder Acknowledgement(s): This research work was supported by the Mississippi INBRE, funded by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20GM103476. Faren White is a Jackson Heart Study Scholar. Jackson Heart Study is supported by contracts HHSN2682018000141, HHSN26820130004C from the National Heart, Lung, and Blood Institute and the National Institute on Minority Health and Health Disparities. The authors thank the participants and data collection staff of the Jackson Heart Study.
Faculty Advisor: Clement Yedjou, email@example.com
Role: I ran the MTT Assay Procedure, Trypan Blue Procedure, and took Microscopic images.