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Effect of miR-662 and miR-24-2-5p overexpressions on gene expression in human MDA-MB-231-luc2-NW1 (NW1) and MCF7 cell lines.

ABSTRACT: MicroRNAs (miRNAs), small non-coding RNAs, are powerful and extremely versatile regulators of the gene expression in cells, playing an important role in every step of cancer progression. Here, we report a list of transcripts regulated by miR-662 and miR-24-2-5p transient overexpressions using miRNA MIMIC transfection (MIMIC-miR-662 and MIMIC-miR-24-2-5p, respectively) compared to control (MIMIC-negCTRL-FAM+) at 36 hours post-transfection in human MDA-MB-231-luc2-NW1 (NW1) (for miR-662 and miR-24-2-5p) and MCF7 (only for miR-24-2-5p) breast cancer cell lines. We found 38 transcripts significantly deregulated (17 downregulated, 21 upregulated) in miR-662-overexpressing NW1 cells compared to mock-transfected cells; 222 (187 downregulated, 35 upregulated) and 34 (32 downregulated, 2 upregulated) transcripts deregulated in miR-24-2-5p-overexpressing NW1 and MCF7 cells, respectively, compared to mock-transfected cells. Furthermore, a total of 30 transcripts (29 downregulated, 1 upregulated) were shared between miR-24-2-5p-overexpressing NW1 and MCF7 cells compared to mock-transfected cells. For miR-662, which we found to promote breast cancer metastasis by stimulating cancer cell stemness (Puppo et al., 2023), we observed the deregulation of Cell Migration Inducing Hyaluronidase 1 (CEMIP), Interleukin 6 receptor (IL6R), High Mobility Group AT-Hook 2 (HMGA2), Small Ubiquitin Like Modifier 3 (SUMO3), and Polymerase (DNA) Delta Interacting Protein 2 (POLDIP2), which reinforced the evidence of the involvement of miR-662 in the acquisition of a stem-like phenotype by human NW1 breast cancer cells and miR-662 contribution to breast cancer bone metastasis formation. For miR-24-2-5p, we found that the downregulation of two transcripts, WD Repeat and FYVE Domain Containing 1(WDFY1) and SH3 Domain Binding Glutamate Rich Protein Like 2 (SH3BGRL2), might explain a possible role of miR-24-2-5p as a tumour suppressor by reducing breast cancer cell invasive properties.

ORGANISM(S): Homo sapiens

PROVIDER: GSE267064 | GEO | 2024/12/27

REPOSITORIES: GEO

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Project description:We tested the hypothesis that a panel of placental mammal-specific miRNAs and their targets play important to establish receptivity to implantation and their dysregulated expression may be a feature in women with early pregnancy loss. Relative expression levels of miR-340-5p, −542-3p, and −671-5p all increased following treatment of Ishikawa cells with progesterone (10 μg/ml) for 24 hrs (p < 0.05). RNA sequencing of these P4-treated cells identified co-ordinate changes to 6,367 transcripts of which 1713 were predicted targets of miR-340-5p, 670 of miR-542-3p, and 618 of miR-671-5p. Quantitative proteomic analysis of Ishikawa cells transfected with mimic or inhibitor (48 hrs: n=3 biological replicates) for each of the P4-regulated miRNAs was carried out to identify targets of these miRNAs. Excluding off target effects, mir-340-5p mimic altered 1,369 proteins while inhibition changed expression of 376 proteins (p < 0.05) of which, 72 were common to both treatments. A total of 280 proteins were identified between predicted (mirDB) and confirmed (in vitro) targets. In total, 171 proteins predicted to be targets by mirDB were altered in vitro by treatment with miR-340-5p mimic or inhibitor and were also altered by treatment of endometrial epithelial cells with P4. In vitro targets of miR-542-3p identified 1,378 proteins altered by mimic while inhibition altered 975 a core of 200 proteins were changed by both. 100 protein targets were predicted and only 46 proteins were P4 regulated. miR-671-mimic altered 1,252 proteins with inhibition changing 492 proteins of which 97 were common to both, 95 were miDB predicted targets and 46 were also P4-regulated. All miRNAs were detected in endometrial biopsies taken from patients during the luteal phase of their cycle, irrespective of prior or future pregnancy outcomes Expression of mir-340-5p showed an overall increase in patients who had previously suffered a miscarriage and had a subsequent miscarriage, as compared to those who had infertility or previous miscarriage and subsequently went on to have a life birth outcome. The regulation of these miRNAs and their protein targets regulate the function of transport and secretion, and adhesion of the endometrial epithelia required for successful implantation in humans. Dysfunction of these miRNAs (and therefore the targets they regulate) may contribute to endometrial-derived recurrent pregnancy loss in women.

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Effect of miR-662 and miR-24-2-5p overexpressions on gene expression in human MDA-MB-231-luc2-NW1 (NW1) and MCF7 cell lines.

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