Prognostic Analysis Of CPEB3 In Melanoma And The Down-regulated Expression Of CPEB3 Is Related To The Malignant Progression

Melanoma is the most malignant tumor among skin tumors.Although melanoma biology has been increasingly studied,no prognostic biomarkers have yet been incorporated into clinical protocols.Cytoplasmic polyadenylation element binding protein 3(CPEB3)has been shown to act as a prognostic biomarker in several cancers.Here,we aimed to investigate the predictive value of CPEB3 for the prognosis of cutaneous melanoma patients.Objective1.To evaluate the role of CPEB3 in predicting the prognosis of melanoma;2.To identify the expression of CPEB3 in melanoma;3.To explore the effect of down-regulated CPEB3 expression on the biological effect of melanoma cells.Methods·Bioinformatics stratified analysis of CPEB3 in melanoma prognosis1.The association of CPEB3 expression and clinical pathologic features was performed using The Cancer Genome Atlas(TCGA)data set.2.The relationship between CPEB3 expression and prognosis of melanoma was analysis.·CPEB3 expression in melanoma cells and tissues1.The expression of CPEB3 in melanoma tissue was detected by immunohistochemistry.2.The expression of CPEB3 in melanoma cell lines was detected by qRT-PCR and Western blot.·Effect of down-regulated CPEB3 expression on biological function of melanoma cells1.Construction of plasmid vector of RNA interference specific for CPEB3,the efficiency of silence was verified by qRT-PCR and Western blot.2.The effects of down-regulated CPEB3 expression on the proliferation,apoptosis,migration and invasion of melanoma cells were detected by CCK-8,plate cloning,flow apoptosis,scratch and Transwell experiments.·Effects of miRNA-301b-3p on proliferation of melanoma cells1.Construct target miRNA.The direct targeting relationship between miRNA and CPEB3 in melanoma cells was verified by luciferase assay.2.CCK8 assay was used to detect the effect of miRNA-301b-3p on cell proliferationResults1.Bioinformation analysis showed that the mRNA has a significant correlation between low expression of CPEB3 and higher T staging(P<0.001),clinical stage(P<0.05),Clark level of melanoma(P<0.05)and ulceration(P<0.05).2.Prognostic analysis showed that low CPEB3 expression was associated with a reduced survival time in melanoma patients(P<0.05).In the univariate analysis,the melanoma Clark level,melanoma ulceration,clinical stage,TNM stage,age and expression of CPEB3 affected the prognosis of melanoma patients(P<0.05).Further analysis by multivariate Cox regression showed that N stage,melanoma ulceration and CPEB3 expression were independent prognostic risk factors of overall survival in melanoma patients(P<0.001).3.Immunohistochemical results showed that the expression of CPEB3 was different in different stages of melanoma(P<0.05).With the increase of malignant stage of melanoma,the expression of CPEB3 tended to be down-regulated.4.The expression of CPEB3 was relatively higher in melanoma cell lines SK-MEL-28 and WM3211,moderate in A2058,and low in A378 and A875.5.Downregulation of CPEB3 promoted melanocyte proliferation,cloning,migration,invasion and metastasis,and inhibit apoptosis(P<0.05).6.In melanoma cell SK-MEL-28,miRNA-301b-3p may accelerate the proliferation of cells by targeting CPEB3.Conclusion1.Low expression of CPEB3 predicted poorer prognosis in melamoma,CPEB3 may be a potential prognostic marker for melanoma;2.CPEB3 may inhibit the development of melanoma.