TCF21 regulates miR-10a-5p/LIN28B signaling to block the proliferation and invasion of melanoma cells

• Published in: PloS one 2021-08, Vol.16 (8), p.e0255971-e0255971
• Main Authors: Zhu, Haijun, Kang, Mengshi, Bai, Xinping
• Format: Article
• Language: English
• Subjects: 3' Untranslated regions; Analysis; Antibodies; Apoptosis; Assaying; Basic Helix-Loop-Helix Transcription Factors - genetics; Basic Helix-Loop-Helix Transcription Factors - metabolism; Bioinformatics; Biology and life sciences; Biomarkers, Tumor - genetics; Biomarkers, Tumor - metabolism; Cell growth; Cell migration; Cell Proliferation; Chromatin; Chromosome 5; Gene expression; Gene Expression Regulation, Neoplastic; Genetic aspects; Health aspects; Humans; Immunoprecipitation; Medicine and Health Sciences; Melanoma; Melanoma - genetics; Melanoma - metabolism; Melanoma - pathology; MicroRNA; MicroRNAs; MicroRNAs - genetics; miRNA; Neoplasm Invasiveness; Pathogenesis; Plasmids; Plastic surgery; Polymerase chain reaction; Prognosis; Prostate cancer; Proteins; Research and Analysis Methods; RNA-Binding Proteins - genetics; RNA-Binding Proteins - metabolism; Skin cancer; Survival Rate; Transcription; Tumor Cells, Cultured
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0255971

Some research has suggested that miRNA-10a (miR-10a-5p) had an inhibitory function in proliferation and invasion of cancers. Whereas the role of miR-10a-5p in melanoma has not been fully explored. This study aims to confirm LIN28B as the targeted gene of miR-10a-5p which was explored in melanoma cells. In addition, upstream regulatory molecule of miR-10a-5p was also investigated in melanoma cells. Real-time Quantitative polymerase chain reaction (RT-qPCR) was adopted to analyze miR-10a-5p expression level in melanoma and the normal human epidermal melanocyte cells. Several biological assays were performed to evaluate miR-10a-5p influences on cell proliferation, migration and invasion ability in A375 and B16-F10 cells. Gene prediction of miRNA targeting and a dual luciferase assay were applied to assess miR-10a-5p-targeted LIN28B. Western blot assessed the impacts of miR-10a-5p on the protein expression of LIN28B. Western blot analyzed the TCF21 effects on the expression of LIN28B and RT-qPCR assessed the influence of TCF21 on the expression level of miRNA-10a. In addition, Chromatin Immunoprecipitation (ChIP) Assay and JASPAR databases were employed to explore the regulatory relationship between TCF21 and miR-10a-5p. We discovered that miR-10a-5p expression was lower in melanoma cells and high expression of miR-10a-5p suppressed the proliferation, migration and invasion abilities of melanoma cells. We also discovered that miR-10a-5p targeted the LIN28B mRNA 3'UTR area and diminished LIN28B protein expression. We found that LIN28B expression was strongly decreased by TCF21 upregulation in the two melanoma cells. The qRT-PCR assay showed that miR-10a-5p expression level was obviously boosted by increased TCF21 expression. The results also demonstrated that TCF21 directly regulated miR-10a-5p at transcript levels. TCF21 induced miRNA-10a targeting LIN28B could affect the progression and growth of melanoma.