MiR-200b is associated with cisplatin sensitivity in bladder cancer cells
ABSTRACT: We aimed to clarify the role of miR-200b in cisplatin (CDDP) sensitivity in bladder cancer (BCa). CDDP resistant T24 cells (T24RC) were transfected with a miR mimic negative control (NC) or a miR-200b mimic, after which cells were treated with or without CDDP. We found that ectopic miR-200b expression re-sensitized the T24RC cells to CDDP. Gene expression microarray analysis revealed that the combination of miR-200b and CDDP affected genes involved in CDDP sensitivity and cytotoxicity. Overall design: CDDP resistant T24 cells (T24RC) were transfected with a miR mimic negative control (NC) or a miR-200b mimic using Lipofectamine RNAiMAX and incubated for 72 h. Transfectants were then treated with or without 1.6 μg/ml of CDDP for additional 72 h. Total RNA was extracted using a TRI Reagent.
INSTRUMENT(S): Agilent-039494 SurePrint G3 Human GE v2 8x60K Microarray 039381 (Feature Number version)
In this study, we identified microRNAs (miRNAs) involved in cisplatin (CDDP) resistance in bladder cancer (BCa). After establishing CDDP-resistant BCa cell lines (T24RC and EJ138RC), TaqMan arrays revealed that members of the miR-200 family (miR-200b, miR-200a and miR-429) were downregulated in T24RC as compared to parental T24 cells. miR-200b was associated with CDDP sensitivity in BCa cells, and its downregulation was associated with CpG island hypermethylation. Pharmacological demethylation u ...[more]
Project description:Genes regulated by miR-206 were identified by microarray analysis in RD cells transfected with a Negative Control (NC) or miR-206 Mimic Overall design: Human RD rhabdomyosarcoma cells were transfected with a Negative Control (NC) miRNA Mimic or a miR-206 miRNA Mimic, 48 hours post transfection RNA was extracted and hybridized on Affymetrix microarrays.
Project description:To identify target genes of cancer-related microRNAs in human cancer, several cell lines (bladder cancer, prostate cancer, renal cell carcinoma, esophageal squamous cell carcinoma, and head and neck squamous cell carcinoma) were subjected to Agilent whole genome microarrays. Human cancer cell lines (BOY, T24, A498, 786-O, caki-1, LNCap, PC3, TE2, T.Tn, FaDu, SAS, HSC3, and IMC-3) were transfected with miRNAs (miR-375, miR-145, miR-200a, miR-200b, miR-200c, miR-141, miR-429, miR-138, miR-218, miR-874, miR-31, miR-222, miR-1285, and miR-206) or siRNAs (si-FOXA1_1, si-FOXA1_3, and si-TAGLN2). The miRNA-transfected human cancer cell lines were compared to control cell lines.
Project description:Microarry analysis of mouse gene expression profile after transfected with miR-27a mimics (27a-7) and mimic NC (NC-9) Goal was to determine the effects of miR-27a transfection on global gene expression. Two-condition experiment, 27a-7 vs.NC-9.
Project description:To investigate genes regulated by miR-489, gene expression analysis was carried out between control siRNA and miR-489 mimic transfected cells. Overall design: T47D cells were transfected with miR-489 mimic or control scramble siRNA for 72 hours. RNA was isolated 72 hours post transfection using trizol reagent. Eacsh sample was collected in triplicate.
Project description:This SuperSeries is composed of the SubSeries listed below. Data for miR-429, miR-205, miR-200b, and miR-141 were each compared independently to the same negative control data contained in the raw .CEL files nc-miR-1-1.CEL, nc-miR-1-2.CEL and nc-miR-1-3.CEL. For each miRNA, the negative control data were re-normalized together with data only from that specific miRNA. The experiment for M12 was performed at a later date and an independent set of negative control data was collected at that time. Therefore, only M12 data was compared to the negative control data contained in the raw .CEL files nc-miR-2-1.CEL, nc-miR-2-2.CEL, and nc-miR-2-3.CEL.
Project description:To strategically identify direct miR-210 target genes, we first performed messenger RNA (mRNA) microarray analyses of MRC5 cells and primary lung fibroblasts that were transiently transfected with miR-210 mimic and the control Overall design: To detect the mRNAs in MRC5 cells and lung fibroblasts treated with miR-210 mimic or pre-miR-NC , 100 ng of total RNA was labelled and hybridized using a SurePrint G3 Human GE v2 8x60K Microarray (Agilent Technologies) according to the manufacturer’s protocol.
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:For the purpose of analyzing mechanisms related to the cis-diamminedichloroplatinum (CDDP) resistance in head and neck squamous cell carcinoma (head and neck SCC), we employed a nasal squamous cell carcinoma (nasal SCC) cell line RPMI2650 and its CDDP resistant substrain RPMI2650CR previously described. The identification of the resistance-related microRNA (miR) clusters was conducted between RPMI2650CR and RPMI2650 using microRNA microarray. microRNA expression of parental and CDDP resistant was measured with or without CDDP treatment in duplicate.
Project description:The extent of intratumoral heterogeneity, the subclonal structures and the mechanisms of treatment-induced clonal selection by cisplatin was investigated in the squamous cell carcinoma cell line model FaDu. We picked 96 single cell-derived clones from the cisplatin-sensitive parental FaDu cell line. After expansion as separate cultures, these clones were tested for their sensitivity to CDDP. By this approach, we isolated individual cell clones that were primarily resistant (clones 5 & 78) and others that showed high sensitivity to CDDP (clones 46 & 54). Basal mRNA expression levels associated with CDDP sensitivity / resistance were determined in two independent microarray analyses.