1.A Case Report Of Acquired Tufted Angioma 2.The Expression Of ThermoTRPs In Melanoma Cells And The Influence Of Menthol And 2-APB On Cell Proliferation And Cell Cycle Of A2058 Cells

Melanoma is a kind of malignant tumor which is derived from melanocytes,and can be found in the skin,eyes,ears and so on.Melanoma,especially when poorly differentiated,is developed rapidly with high invasiveness.Transient receptor potential channel(TRP)is a group of non-selective cation ion channels.To date,28 members of TRP family were found in mammal.According to the homology of the amino acid sequence,these channels can be divided into 6 subfamilies:TRPC(Canonical),TRPV(Vanilloid),TRPM(Melastatin),TRPP(Polycystin),TRPML(Mu colipin)and TRPA(Ankyrin).Several TRPs are sensitive to temperature and can be activated by the temperature threshold respectively.Now researches on TRPA1,TRPM8,TRPV1,TRPV2,TRPV3,TRPV4 are much more than others,and it has been reported that TRPA1 and TRPM8 are expressed in melanoma cells.TRPs,as an ion channel,are permeable of calcium ion which is crucial to many physiology and pathological processes.So it is significant to find out TRPs in melanoma and the difference between melanoma and the normal melanocytes.Objective: To determine and compare the expression of TRPs protein and m RNA in human melanoma cells and normal melanocytes.To investigative the pathophysiologic role of TRPs in melanoma by observing the cell viability and cell cycle after treating the melanoma cell line A2058 with menthol and 2-Aminoethoxydiphenyl borate(2-APB) which are activators of TRPs.Methods: 1.Determine and compare the expression of protein TRPA1,TRPM8,TRPV2 and TRPV4 in the melanoma cells lines SK-MEL-3,A2058,G361,A375 and Primary Epidermal Melanocytes(ATCC?PCS-200-013)by western blotting.2.The m RNA expression of TRPA1,TRPM8,TRPV2 and TRPV4 in the melanoma cells mentioned above and Primary Epidermal Melanocytes(ATCC?PCS-200-013)were tested by real-time PCR.3.The cell viability of melanoma cell A2058 was analyzed by MTT assay after cells were treated with different concentrations of menthol and 2-APB.4.The cell cycle of melanoma cell A2058 was analyzed by flow cytometry after cells were treated with different concentrations of menthol and 2-APB.Results: 1.Combined the results of western blotting and q PCR,the expression of TRPA1,TRPM8,TRPV2 and TRPV4 was detected in tested melanoma cells.Compared with Primary Epidermal Melanocytes(ATCC?PCS-200-013),the expression levels of protein and m RNA in melanoma cells were up-regulated in terms of TRPA1,TRPM8 and TRPV4,whereas TRPV2 showed a downward trend.2.MTT assay showed that when cultured with menthol or 2-APB,the cell proliferation was inhabited at 1m M(or above)and 0.2m M(or above)respectively(P<0.05).3.When observed by microscope,the proliferation of cell A2058 was inhibited distinctly when treated with 0.2m M 2-APB for 48 hours.4.When cultured with menthol for 12 hours and 48 hours,no significant difference had been shown in the test,while treated with 0.4 m M 2-APB for 48 hours,S phase of A2058 cells increased compared with wild type.Conclusions: 1.There was the expression of TRPA1,TRPM8,TRPV2 and TRPV4 in melanoma cells.2.Compared with melanocytes the expression of TRPA1,TRPM8 and TRPV4 was up-regulated and the expression of TRPV2 was down-regulated in melanoma cells.3.In melanoma cell line A2058,menthol can inhibit its proliferation without any influence to cell cycle,while 2-APB(the activator of TRPs)can block its cell cycle in S phase.