Evaluation of the effect of anti-mir21 on apoptosis of class A-375 melanoma

Abstract

Cancer includes a group of diseases whose main characteristic is unregulated cell growth, invasion and spread of cells from the primary site to other parts of the body. Despite recent advances in diagnosis and treatment, cancer is one of the most important causes of death in the world. In addition to genetic and environmental factors, epigenetic factors are involved in the etiology of cancer. Cancer is the third cause of death in Iran after heart problems and driving accidents, the death caused by cancer has been increasing in recent decades; Apoptosis is the process of programmed cell death. It is used in the early stages of growth to eliminate unwanted cells. In adults, apoptosis is used to protect the body from cells that are irreparably damaged. It has also been proven that apoptosis plays an effective role in preventing the spread of cancer. This study also aims to investigate the success rate of targeting miRNA-21 to prevent the development and spread of melanoma; Therefore, the aim of this study is to investigate the effect of anti-mir21 on apoptosis of A-375 melanoma. Materials and methods: First, A-375 melanoma cells are cultured in RPMI culture medium containing 10% FCS at 37ₒc and 5% CO2, then the anti-mir21 gene is added to the culture medium along with the vector and transfecting agent Lipofectamin2000. Incubation is done, the rate of growth and proliferation of cancer cells is checked by MTT technique. After transfecting anti-mir21 and passing the necessary time, the culture medium is replaced with culture medium containing MTT and after 4 hours of incubation, light absorption is read by ELISA plate reader at 570 nm wavelength. The rate of migration and metastasis of cancer cells after transfection is checked by wound healing assay. 5x105 cells transfected with MIR-21 in the incubated culture medium, a groove is created in the medium, after washing the medium with PBS, incubation is done, and then the groove location is photographed by a microscope, and the migration of cells from the edge of the gap It is checked inside. The amount of apoptosis is also done using flow cytometry technique of Annexin 5 and PI. Results: Proliferation of cancer cells decreased after exposure to encomirs, and metastasis of melanoma cells decreased after exposure to encomirs.