The Infuction Of CD147 In Involved In The Apoptosis Of Human Malignant Melanoma Cells G361 By Resveratrol

ObjectiveTo investigated the effect of resveratrol on growth and apoptosis in human malignant melanoma cell line G361 and the role of CD147 in it.Methodsl.The human malignant melanoma cell line G361 was treated by resveratrol (sigma) with varying concentrations after 48h.Cell growth was assessed by using MTT assay.2.The human malignant melanoma cell line G361 was treated by resveratrol (sigma) with varying concentrations after 48h, the expression of XIAP,Caspase-9 and Caspase-3 proteins was assessed by western blot.3.The human malignant melanoma cell line G361 was treated by resveratrol (sigma) with varying concentrations after 48h,the expression of CD 147 was assessed by reverse transcriptase-polymerase chain reaction (RT-PCR) and western blot.4.We previously designed PcDNA3.0 sequences that target human malignant melanoma cell line G361 CD147mRNA, and transfected with empty vector as control,the cells was treated or wasn't treated by resveratrol, the expression of CD 147, XIAP, Caspase-9 and Caspase-3 proteins was assessed by western blot.6.Flow cytometric analysis and electron microscopic observation revealed differential cell apoptotic status related to CD147 expression.Results1.Resveratrol inhibited cell proliferation in a concentration -dependent manner by MTT assay,and then we could acquire suitable concentration by calculated IC50, IC50=21.06±0.383μM2.G361 cells were cultured with different concentrations of resveratrol of 48h,XIAP expression was down-regulated while caspase-3 and caspase-9 were up-regulated.This indicate resveratrol could induces the the apoptosis of G361 cells.3.CD 147 production was significantly inhibited by resveratrol,Compared with control,RT-PCR analysis showed that CD147 expression level in treated cells by resveratrol with vary concentration(2μM,4μM,8μM,16μM)were 62.82%,56.27%,50.27%,43.25% (p<0.05).Western blot analysis CD147 production was significantly inhibited by resveratrol in concentration-dependent but not in time-dependent manner(p>0.05).4.Compared with control, the expression of CD 147 and XIAP was up-regulated while Caspase-9 and Caspase-3 were down-regulated in G361 cells of transfected CD147cDNA. When expression of CD 147 was down-regulated by resveratrol, XIAP was down-regulated and Caspase-9 and Caspase-3 were up-regulated.These findings also revealed that resveratrol induces the apoptosis of G361 cells by down-regulated expression of CD147.5. G361 cells of transfected CD147cDNA was treated by resveratrol, The average % apoptosis was 2.38%, the numbe was significantly higher than those of in control and G361 cells of transfected CD147cDNA (p< 0.05), The average % apoptosis in G361 cells of transfected CD147cDNA (0.30%) was lower than those of control (0.76%).This findings indicated that CD147 expression could effectively protect G361 cells from apoptosis.Resveratrol induces the apoptosis of G361 cells by down-regulated expression of CD 147.6. G361 cells of transfected CD147cDNA was treated by resveratrol, the cells was small and round,the typical morphology of apoptosis such as condensation and fragmentation of nuclei and blebbing of membrane were observed.Conclusion1.Resveratrol inhibits cell growth and induces the apoptosis of the human malignant melanoma cell line G361.2.Resveratrol inhibits the expression of CD147 in a dose-dependent manner but not in a time-dependent.3.CD 147 expression could effectively protect of the human malignant melanoma cell line G361 from apoptosis, resveratrol induces the apoptosis of G361 cells by down-regulated expression of CD147.