HSP70 effect on HMGA1 and HMGB1 in Melanoma cell line (A-375)
Abstract
The aim of this study was to determine the effect of HSP70 on HMGA1, HMGB1 in melanoma cell class (A-375).
Methods: The present laboratory study was performed on melanoma cancer cells in RPMI1640 culture medium containing 10% FCS at 37 ₒ C and 5% CO2. After counting and dividing the cells, the inside of the cell plate was stimulated as a time course with HSP70 protein. Incubation was performed. After 48 hours, the cells were collected and total mRNA was extracted and the expression of HMGA1 and HMGB1 expression was investigated through qRT-PCR. The supernatant of cells stimulated by HSP70 was collected and measured with an Elisa interleukin 6 kit, IL6 level. The rate of growth and proliferation of cancer cells was assessed by the MTT technique. The collected information was analyzed using Student t-test and SPSS software and was considered significant in all P <0.05 statistical tests.
Results: The results of the present study indicated that the expression of HMGA1 and HMGB1 proteins, 24 hours after stimulation of melanoma cell line (A-375) with HSP70, decreased significantly (P <0.001). This decrease started 12 hours after stimulation. The expression levels of TLR2 and TLR4 in melanoma cell line (A-375) were also significantly reduced in exposure to HSP70. The penetration of IL17 and IL6 into melanoma cells increased with exposure to HSP70.