Project description:In this experiment, we want to assess the effect of a lentiviral miR-10a and miR-335 overexpression on the transcriptome of murine LSK (Lin-,Sca-1+,c-Kit+) cells. Primary LSK cells were transduced with lentiviral miRNA overexpression constructs (control: GFP overexpression) and sorted for transduced cells (GFP+) after five days of in vitro culture (Flt-3, TPO, IL-3, SCF containing media).
Project description:To clarify the effect of miRNAs, we carried out a gene expression microarray analysis using GIST-T1 cells transfected with a miR-335 mimic or a negative control. We found that 1,095 probe sets (853 unique genes) were downregulated (>1.5-fold) by ectopic miR-335 expression.
Project description:To clarify the effect of miRNAs, we carried out a gene expression microarray analysis using GIST-T1 cells transfected with a miR-335 mimic or a negative control. We found that 1,095 probe sets (853 unique genes) were downregulated (>1.5-fold) by ectopic miR-335 expression. GIST-T1 cells were transfected with a mirVana miR-335 mimic (Ambion) or a mirVana miRNA mimic Negative Control #1 (Ambion). Forty-eight hours after transfection, total RNA extraction was carried out, and gene expression signatures were analyzed.
Project description:Prenatal overexposure to glucocorticoids (GC) can lead to cognitive impairment in offspring, yet the underlying mechanisms remain elusive, and effective clinical interventions are still lacking. In this study, we analyzed the miRNA expression profile in hippocampal neural stem cells (NSCs) from gestational synthetic GC analog dexamethasone (DEX) exposure (GDE) offspring mice, and found that miR-335-3p expression was significantly downregulated. Functional experiments demonstrated that miR-335-3p specifically reverses the inhibitory effect of DEX on NSC proliferation without affecting their differentiation capacity. Mechanistic studies revealed that DEX activates the glucocorticoid receptor (GR) to directly suppress miR-335-3p transcription, which in turn relieves its targeted inhibition of PTEN, ultimately impairing NSC proliferation and cognitive function. This DEX/miR-335-3p/PTEN regulatory axis is highly conserved in human embryonic stem cells (hESCs)-derived 2D NSCs and 3D cerebral organoids. Importantly, intrahippocampal delivery of miR-335-3p agomir effectively restored NSC proliferation and ameliorated learning and memory deficits in GDE offspring mice. These findings identify a previously unrecognized miRNA-mediated epigenetic axis underlying prenatal GC exposure-induced cognitive impairment in offspring. Our study provides mechanistic insights into the developmental origins of glucocorticoid-induced neurological disorders and highlights miR-335-3p as a potential therapeutic target for preventing fetal-origin cognitive deficits.
Project description:the LM2 breast cancer cell line is an in vivo derived line from the MDA-MB-231 parental line. this LM2 line has been transduced either with a short hairpin control or miR-335 expression vector. Experiment Overall Design: the LM2 cell line is transduced either with a short hairpin control vector or miR-335 expression vector.
Project description:the LM2 breast cancer cell line is an in vivo derived line from the MDA-MB-231 parental line. this LM2 line has been transduced either with a short hairpin control or miR-335 expression vector. Keywords: breast cancer, metastasis, miRNA