Analysis Of Candidate Genes Expected To Be Essential For Melanoma Surviving

Abstract

BackgroundNew approach for treating cancer is to influence on vital genes for tumors. A number of attempts have led to discovery candidate genes that are mean to be essential for surviving of tumor cells of different types. In this work, we tried to analyze genes that were previously found to be essential for melanoma surviving. We performed transient siRNA-mediated knockdown of UNC45A, STK11IP, RHPN2 and ZNFX1 in melanoma cell line A375. After that, we checked cell viability, proliferation and migrating rate in vitro.MethodsFor knockdown of the genes we used siRNA mediated interference. To deliver siRNA in cells we used transfection by Metafectene PRO. Cell viability and proliferation assessed by MTT assay. Cell migration was assessed by wound healing assay. Knockdown of genes was evaluated via RT-qPCR analysis.ResultsOur results indicate that knockdown of the genes leads to no statistically significant changes in cell survival according to viability assay. However, mobility assay shows that knockdown of each of the target genes accelerates the speed of cells migrating.ConclusionOur results do not confirm the hypothesis. Possible explaining of such controversial results could be in insufficient bioinformatical analysis or in need of multiplex knockdown of the genes.

A myosin chaperone, UNC‐45A, is a novel regulator of intestinal epithelial barrier integrity and repair

“…Depletion of this chaperone was shown to inhibit motility of breast and ovarian cancer cells, 27,29 but promoted migration of osteosarcoma and melanoma cells. 30,62 This resembles the reported opposite effects of NM-II inhibition on cell migration under different experimental conditions 50,63 and may reflect complex regulatory effects of UNC-45A-driven myofilament assembly in motile cells. We observed that loss of UNC-45A inhibited both collective migration of IEC sheets and individual cell passage through membrane pores (Figure 5A-H).…”

“…Generally, the roles of UNC‐45A in mammalian cell migration remain controversial. Depletion of this chaperone was shown to inhibit motility of breast and ovarian cancer cells, 27,29 but promoted migration of osteosarcoma and melanoma cells 30,62 50,63 and may reflect complex regulatory effects of UNC‐45A‐driven myofilament assembly in motile cells.…”