Project description:miR-125b-5p is a well known miRNA already describded in several forms of cancer. miR-125b-5p is expressed in adipose tissue, adipocytes as well as their precursor cells. We aim to invest the role of miR-125b-5p in white adipocytes conversion into brite adipocytes. To get an idea about putative targets of miR-125b-5p in adipocyte conversion, we transfected miR-125b-5p mimic in human Multipotent Adipose-Derived Stem (hMADS) cells, differenciated in white adipocytes. Gene expression profiling is performed 48h after hMADSC transfection. Two-condition experiment, hMADS cells at day 16 after conversion of white adipocytes into brite adipocytes, comparison of cells transfected with a mimic miR-125b-5p to cells transfected with a negative controle. Biological replicates: 4, indepently grown and harvested. On each array, one biological replicate of mimic miR-125b-5p transfected cells was directly compared to one biological replicate of mimic negative control transfected cells (serving as reference sample). All hybridizations were repeated with reversed dye assignment (dye-swap) as technical replicates.
Project description:miR-125b-5p is a well known miRNA already describded in several forms of cancer. miR-125b-5p is expressed in adipose tissue, adipocytes as well as their precursor cells. We aim to invest the role of miR-125b-5p in white adipocytes conversion into brite adipocytes. To get an idea about putative targets of miR-125b-5p in adipocyte conversion, we transfected miR-125b-5p mimic in human Multipotent Adipose-Derived Stem (hMADS) cells, differenciated in white adipocytes. Gene expression profiling is performed 48h after hMADSC transfection.
Project description:Post-transcriptional regulation of gene expression by miRNAs likely makes significant contributions to mRNA abundance at the embryo-maternal interface. In this study, we investigated how miR-26a-5p and miR-125b-5p contribute to molecular changes occurring in the uterine luminal epithelium, which serves as the first site of signal exchange between the mother and developing embryo. To measure de novo protein synthesis after miRNA delivery to primary uterine luminal epithelial cells, we employed pulsed stable isotope labeling by amino acids (pSILAC). We found that both miRNAs alter the proteome of luminal epithelial cells, impacting numerous cellular functions, immune responses, as well as intracellular and second messenger signaling pathways. Additionally, we identified several features of miRNA-mRNA interactions that may influence the targeting efficiency of miR-26a-5p and miR-125b-5p. Overall, our study suggests a complex interaction of miR-26a-5p and miR-125b-5p with their respective targets. However, both appear to cooperatively function in modulating the cellular environment of the luminal epithelium, facilitating the morphological and molecular changes that occur during the intensive communication between the embryo and uterus at pregnancy.
Project description:To investigate machanism of miR-423-5p regulating the angiogenic ability of bEnd.3 cells, we transfected miR-423-5p mimic to overexpress miR-423-5p in bEnd.3 cells. Then we performed high throughput sequencing of miR-423-5p mimic-transfected and control bEnd.3 cells to evaluate different gene expressions between miR-210-3p-overexpressing and control.
Project description:Transcriptional profiling of breast cancer cells comparing pre-control transfected cells with cells transfected with pre-miR-125b. We searched for miR-125b targets by systematic screening of mRNA profiling of pre-miR-125b transfected MCF-7 cells and MDA-MB-435 cells.
Project description:This experiment is designed to investigate the impact of exosomal miR-145-5p on the functional pathway and molecules on RPTEC cells. Data-independent acquisition (DIA) proteomics was conducted in the RPTEC cells transfected with miR-145-5p mimic or scramble control.
Project description:To investigate the gene expresiion regulated by miR-125b and its isomiRs. We transfected H9C2 cells with respective mimics and performed gene expression profiling anlaysis using data from RNA-seq of mimic controls, miR-125b and its isomiRs mimics subjected to normoxia and hypoxia treatment.
Project description:Patients with advanced colorectal cancer (CRC) are commonly treated with systemic combination therapy but suffer eventually from drug resistance. MicroRNAs (miRNAs) are suggested to play a role in treatment resistance of CRC. We studied whether restoring downregulated miR-195-5p and 497-5p sensitize CRC cells to currently used chemotherapeutics 5-fluorouracil, oxaliplatin and irinotecan. Sensitivity to 5-FU, oxaliplatin and irinotecan before and after transfection with miR-195-5p and miR-497-5p mimics was analyzed in CRC cell lines HCT116, RKO, DLD-1 and SW480. Mass spectrometry based proteomic analysis of transfected and wild-type cells was used to identify targets involved in sensitivity to chemotherapy. Proteomic analysis revealed 181 proteins with significantly altered expression after transfection with miR-195-5p mimic in HCT116 and RKO, including 118 downregulated and 63 upregulated proteins. After transfection with miR-497-5p mimic, 130 proteins were significantly downregulated and 102 were upregulated in HCT116 and RKO (P<0.05 and FC<-3 or FC>3). CHUK and LUZP1 were coinciding downregulated proteins in sensitized CRC cells after transfection with either mimic. Resistance mechanisms of these two proteins may be related to nuclear factor kappa-B signaling and G1 cell cycle arrest, respectively. Restoring miR-195-5p and miR-497-5p expression enhanced sensitivity to chemotherapy, mainly oxaliplatin, in CRC cells and could be a promising treatment strategy for patients with mCRC. Proteomics revealed potential targets of these miRNAs involved in sensitivity to chemotherapy.
Project description:Transcriptional profiling of breast cancer cells comparing pre-control transfected cells with cells transfected with pre-miR-125b. We searched for miR-125b targets by systematic screening of mRNA profiling of pre-miR-125b transfected MCF-7 cells and MDA-MB-435 cells. Two-condition experiment, pre-miR-125b Transfected vs. pre-control Transfected MCF-7 cells. One replicate per array.
Project description:Illumina bead array data from TCam2, 1411H, and 2102Ep cell lines for mRNA expression. Cell lines were treated with either mismatch negative control (MNC) or combination miRNA replenishment with equimolar concentration of miR-100-5p and miR-125b-5p. Five comparisons are provided; namely TCam2 cell line on day 2 (d2 )and d7; 1411H cell line on d2 and d7; and 2102Ep cell line on d7. From the Materials and Methods section of the formal publication: We undertook global mRNA microarray profiling of TCam2, 1411H, and 2102Ep cells, comparing cells treated with combination miR-100-5p and miR-125b-5p mimics (total concentration 16.7 nm) with controls. At d7 after treatment, 24 triplicate samples were profiled, comprising TCam2, 1411H, and 2102Ep cell lines receiving combination treatment (n = 9 samples) and MNC treatment (n = 9), plus untreated cells for TCam2 and 1411H (n = 6). Importantly, no substantial differences in mRNA gene expression were observed when comparing the d7 MNC and respective untreated cells, highlighting the suitability of MNC-treated cells to act as the control comparison for mimic-treated cell conditions (data available at EBI Array Express, see below). At d2 after treatment, 12 triplicate samples were profiled, comprising TCam2 and 1411H cells receiving combination treatment (n = 6) and MNC treatment (n = 6). 2102Ep cells were excluded from d2 global mRNA profiling due to the absence of phenotypic effects at d7 post-replenishment.