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MIR133A regulates cell proliferation, migration, and apoptosis by targeting SOX9 in human colorectal cancer cells.

ABSTRACT: The human microRNA 133A (MIR133A) was identified as a CRC-associated miRNA. It was down-regulated in human CRC tissues. We identified the putative MIR133A1 and A2 target genes by comparing the transcriptome analysis data of MIR133A1 and A2 knock-in cells with the candidate MIR133A target genes predicted by bioinformatics tools. We identified 29 and 33 putative MIR133A and A2 direct target genes, respectively. Among them, we focused on the master transcription regulator gene SRY-box transcription factor 9 (SOX9), which exhibits a pleiotropic role in cancer. We confirmed that SOX9 is a direct target gene of MIR133A by luciferase reporter assay, quantitative RT-PCR, and western blot analysis. Overexpression of MIR133A in CRC cell lines significantly decreased SOX9 and its downstream PIK3CA-AKT1-GSK3B-CTNNB1 and KRAS-BRAF-MAP2K1-MAPK1/3 pathways and increased apoptosis. Furthermore, functional studies reveal that cell proliferation, colony formation, and migration ability were significantly decreased by MIR133A-overexpressed CRC cell lines. Knockdown of SOX9 in CRC cell lines by SOX9 gene silencing showed similar results. We also used a xenograft model to show that MIR133A overexpression suppresses tumor growth and proliferation. Our results suggest that MIR133A regulates cell proliferation, migration, and apoptosis by targeting SOX9 in human colorectal cancer.

SUBMITTER: Lamichhane S

PROVIDER: S-EPMC9360235 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature

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MIR133A regulates cell proliferation, migration, and apoptosis by targeting SOX9 in human colorectal cancer cells.

Lamichhane Santosh S Mo Ji-Su JS Sharma Grinsun G Joung Sun-Myoung SM Chae Soo-Cheon SC

American journal of cancer research 20220715 7

The human microRNA 133A (MIR133A) was identified as a CRC-associated miRNA. It was down-regulated in human CRC tissues. We identified the putative MIR133A1 and A2 target genes by comparing the transcriptome analysis data of MIR133A1 and A2 knock-in cells with the candidate MIR133A target genes predicted by bioinformatics tools. We identified 29 and 33 putative MIR133A and A2 direct target genes, respectively. Among them, we focused on the m ...[more]

PMID: 35968353

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Project description:Curcumin is the main component of the Chinese herbal plant turmeric, which has been demonstrated to possess antitumor and other pharmacological properties. The aim of the present study was to investigate the effects of curcumin on the viability, migration and apoptosis of human colorectal carcinoma HCT?116 cells, and to explore the underlying molecular mechanisms. In addition, it was investigated whether the antitumor effect of curcumin on HCT?116 cells could match that of the chemotherapeutic drug 5?fluorouracil (5?FU). HCT?116 cells were treated with curcumin (10, 20 and 30 µM) and 5?FU (500 µM), and cell viability and proliferation were detected by Cell Counting Kit?8 and colony formation assays, respectively. The migration and invasion of treated cells were determined using Transwell and carboxyfluorescein succinimidyl amino ester fluorescent labeling assays. Cell cycle distribution and apoptosis rates were detected by flow cytometry. Furthermore, cell morphology changes associated with apoptosis were observed by fluorescence microscopy with acridine orange/ethidium bromide dual staining. To investigate the possible underlying molecular mechanisms, the gene and protein levels of Fas, Fas?associated via death domain (FADD), caspase?8, caspase?3, matrix metalloproteinase (MMP)?9, nuclear factor (NF)??B, E?cadherin and claudin?3 were detected using quantitative PCR analysis, zymography and western blotting. The results revealed that curcumin markedly inhibited the viability and proliferation of HCT?116 cells in a dose? and time?dependent manner. The migration, aggregation and invasion of HCT?116 cells into the lungs of mice were decreased by curcumin treatment in a dose?dependent manner. S?phase arrest and gradually increased apoptotic rates of HCT?116 cells were observed with increasing curcumin concentrations. Additionally, the mRNA and protein levels of apoptosis?associated proteins (Fas, FADD, caspase?8 and caspase?3) and E?cadherin in HCT?116 cells were upregulated following treatment with curcumin in a dose?dependent manner. By contrast, the expression of migration?associated proteins, including MMP?9, NF??B and claudin?3, was downregulated with increasing curcumin concentrations. These data suggested that the inhibitory effect of curcumin on HCT?116 cells may match that of 5?FU. Therefore, curcumin induced cell apoptosis and inhibited tumor cell metastasis by regulating the NF??B signaling pathway, and its therapeutic effect may be comparable to that of 5?FU.

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MIR133A regulates cell proliferation, migration, and apoptosis by targeting SOX9 in human colorectal cancer cells.

Publications

<i>MIR133A</i> regulates cell proliferation, migration, and apoptosis by targeting SOX9 in human colorectal cancer cells.

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