Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Transcriptomic-based identification of miR-125a novel targets in human hepatocarcinoma cells

ABSTRACT: The experimental work unveils novel targets of hsa-miR-125a-5p relevant in hepatocellular carcinoma (HCC). In particular, a genome-wide perspective of the whole miR-125a targetome has been achieved. First, two different HCC cell lines were subjected to a miRNA boosting by mimic transfections, resulting in many de-regulated genes, as observed by a transcriptomic approach. Then, the merging of down-regulated genes with results from bioinformatic predictive tools yielded a number of miR-125a direct targets candidates. Finally, those potential targets were further experimentally validated. This study could pave the way to piece together the RNA regulatory networks governed by miR-125a impacting on hepatocarcinogenesis and be exploited in the future for identifying novel biomarkers and therapeutic targets in HCC.

INSTRUMENT(S): NextSeq 550

ORGANISM(S): Homo sapiens

SUBMITTER: Ilenia De Leo

PROVIDER: E-MTAB-14731 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The ATP binding cassette (ABC) transporter family is widely distributed in vertebrates and is essential for drug resistance, cell signaling and energy homeostasis. There is growing evidence that various ABC transporters contribute to the growth and development of tumors but relatively little is known about how the ABC transporter family behaves in hepatocellular carcinoma (HCC). ABCC6 transporter was downregulated in HCC tissues and that it was associated with successful treatment for HCC patients. Cellular model studies have shown that ABCC6 plays a role in the migration and cytoskeleton rearrangement of HepG2 hepatocarcinoma cells, highlighting its role in cancer biology. Abcc6-silenced HepG2 cells are used as cell model to obtain more deep information about the molecular mechanisms underlying the observed results. MTT and colony formation assays, showed the effects of Abcc6 on HepG2 cell proliferation. Western blotting analysis, real-time PCR, and immunofluorescence were used to find the E-cadherin, Vimentin, and N-cadherin markers associated with the epithelial-to-mesenchymal transition (EMT). Colony formation experiments in the current study showed that Abcc6 decreased HepG2 cell viability. The migratory and invasion activities were dramatically slowed down by Abcc6 silencing, according to the Transwell and wound-healing assays. In tumor cells, EMT has been shown to be crucial for enhancing migration and invasion and is frequently characterized by a loss of epithelial markers (E-cadherin) and an increase in mesenchymal markers (Vimentin and N-cadherin). In the western blotting examination, E-cadherin expression was considerably elevated compared to the control group, while N-cadherin and Vimentin expression were downregulated. This led to the hypothesis that the underlying mechanism of Abcc6 knockdown prevents migration and invasion in HepG2 cells and is linked to the suppression of EMT. In conclusion all evidence suggested that ABC transporters play a more active role in cancer biology.

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Transcriptomic-based identification of miR-125a novel targets in human hepatocarcinoma cells

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