The effect of Leishmania major lysate on the expression of molecular markers of metastasis and angiogenesis of skin cancer cell line

Abstract

There is evidence that contamination with infectious and parasitic agents in a non-specific way can have an effect on the growth of malignant cells. Biological methods are among the promising and healthy methods in the field of cancer treatment, which have received a lot of attention today. Recent studies have shown that the antigens of some parasites have an anti-cancer effect. Therefore, in this study, the effect of Leishmania major parasite lysate on the growth of skin cancer cells was investigated. Methods: skin cancer cell line in RPMI1640 culture medium containing 10% FBS, 2 mM L-glutamine, penicillin (100 Unit/ml) and streptomycin (100 Unit/ml), in sterile conditions under 37 degrees temperature and 5% CO2 Cultivated. Then these cells were treated with different concentrations of Leishmana major lysate; to obtain the optimal concentration(s), the MTT test was performed. In this way, in 96-well plates, in different wells, a certain number of cells were adjacent to different concentrations of antigen, and after 24 hours of incubation, we removed the supernatant solution and added complete medium and MTT to the wells. After 4 hours of incubation at 37°C and then adding MTT solvent, the result was reported based on the color produced and measured by the Elizarider device at a wavelength of 570 nm; Then the cells were added to the number of 400,000 in each well of the 6-well culture plate and after 24 hours when the cells were attached to the bottom of the well, the culture medium was changed and the optimal concentration of antigen was added to them. After 24 hours, the cells were removed with trypsin-EDTA and their total RNA was extracted and cDNA was synthesized using the extraction kit, then the mRNA expression level of angiogenic genes was checked along with the beta-actin internal control by qRT-PCR method and with specific primers Results: Based on the results of the experiments, it was found that angiogenesis in melanoma cell line increased significantly with the help of Leishmania parasite..