Guili Fu1, Jingjing Lu, Yuanquan Zheng, Pan Wang, Qin Shen. 1. Department of Dermatology, Wuhan Children's Hospital (Wuhan Maternal and Child Healthcare Hospital), Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Abstract
PURPOSE: To study the expression of micro ribonucleic acid-320a (miR-320a) in melanoma cells and its influence on the biological functions of these cells. METHODS: MiR-320a expression data and clinical data in melanoma tissues were downloaded from The Cancer Genome Atlas (TCGA) database. Real time-quantitative polymerase chain reaction (RT-qPCR) was used to detect miR-320a expression in melanoma tissues, malignant melanoma cell lines (A375, SKMEL-28 and A2058) and human epidermal melanoma (HEM) cells. The miR-320a mimic was transfected into A375 cells, and the functions of cells were detected. The luciferase reporter gene assay was employed to verify the miR-320a downstream target protein predicted by the biological information prediction software. RESULTS: The differential analysis of miRNAs in melanoma tissues from TCGA database showed that miR-320a expression in melanoma tissues was significantly lower than that in adjacent tissues, and low expression of miR-320a exhibited a severe poor prognosis (p<0.01). MiR-320a mimic could significantly enhance the expression level of miR-320a (p<0.01). The absorbance at 490 nm of A375 cells overexpressing miR-320a decreased remarkably and their proliferation ability was weakened (p<0.01). Overexpression of miR-320a in A375 cells inhibited cell migration to wound parts and epithelial-mesenchymal transition (EMT), invading malignant phenotype (p<0.05). Flow cytometry was employed and it was denoted that after transfecting with miR-320a, the apoptosis rate of A375 cells was elevated overtly (p<0.01). The dual luciferase report test indicated that the luciferase activity in wild type pre-B-cell leukemia transcription factor 3 (PBX3) was markedly lower than that of mutant type PBX 3 (p<0.05). CONCLUSIONS: Targeted binding of miR-320a to PBX3 protein can inhibit the malignant phenotype of cells and affects the occurrence and development of melanoma.
PURPOSE: To study the expression of micro ribonucleic acid-320a (miR-320a) in melanoma cells and its influence on the biological functions of these cells. METHODS:MiR-320a expression data and clinical data in melanoma tissues were downloaded from The Cancer Genome Atlas (TCGA) database. Real time-quantitative polymerase chain reaction (RT-qPCR) was used to detect miR-320a expression in melanoma tissues, malignant melanoma cell lines (A375, SKMEL-28 and A2058) and human epidermal melanoma (HEM) cells. The miR-320a mimic was transfected into A375 cells, and the functions of cells were detected. The luciferase reporter gene assay was employed to verify the miR-320a downstream target protein predicted by the biological information prediction software. RESULTS: The differential analysis of miRNAs in melanoma tissues from TCGA database showed that miR-320a expression in melanoma tissues was significantly lower than that in adjacent tissues, and low expression of miR-320a exhibited a severe poor prognosis (p<0.01). MiR-320a mimic could significantly enhance the expression level of miR-320a (p<0.01). The absorbance at 490 nm of A375 cells overexpressing miR-320a decreased remarkably and their proliferation ability was weakened (p<0.01). Overexpression of miR-320a in A375 cells inhibited cell migration to wound parts and epithelial-mesenchymal transition (EMT), invading malignant phenotype (p<0.05). Flow cytometry was employed and it was denoted that after transfecting with miR-320a, the apoptosis rate of A375 cells was elevated overtly (p<0.01). The dual luciferase report test indicated that the luciferase activity in wild type pre-B-cell leukemia transcription factor 3 (PBX3) was markedly lower than that of mutant type PBX 3 (p<0.05). CONCLUSIONS: Targeted binding of miR-320a to PBX3 protein can inhibit the malignant phenotype of cells and affects the occurrence and development of melanoma.