Journal: Journal of the American Society of Nephrology : JASN

Article Title: miR-335 and miR-34a Promote Renal Senescence by Suppressing Mitochondrial Antioxidative Enzymes

doi: 10.1681/ASN.2010040367

Figure Lengend Snippet: The significantly differentially expressed miRNAs in aging kidney

Article Snippet: LC Sciences (Houston, TX) provided the miRNA microarray chip (μParaflo microfluidic chip).

Techniques:

Journal: Journal of the American Society of Nephrology : JASN

Article Title: miR-335 and miR-34a Promote Renal Senescence by Suppressing Mitochondrial Antioxidative Enzymes

doi: 10.1681/ASN.2010040367

Figure Lengend Snippet: miR-184, miR-335 and miR-347 exhibited differential expressions in (A) aged rats and (B) mice. (A) Fold differences between old and young rat kidneys measured by chip and qRT-PCR. (B) Expression levels of mmu-miR-184, mmu-miR-335, and mmu-miR-347 in the aged mouse kidneys detected by qRT-PCR. n = 5 per miRNA. *P < 0.01 versus young.

Article Snippet: LC Sciences (Houston, TX) provided the miRNA microarray chip (μParaflo microfluidic chip).

Techniques: Quantitative RT-PCR, Expressing

Journal: Journal of the American Society of Nephrology : JASN

Article Title: miR-335 and miR-34a Promote Renal Senescence by Suppressing Mitochondrial Antioxidative Enzymes

doi: 10.1681/ASN.2010040367

Figure Lengend Snippet: Antioxidation-related miRNAs differentially expressed in old kidney and their target genes

Article Snippet: LC Sciences (Houston, TX) provided the miRNA microarray chip (μParaflo microfluidic chip).

Techniques:

Journal: Journal of the American Society of Nephrology : JASN

Article Title: miR-335 and miR-34a Promote Renal Senescence by Suppressing Mitochondrial Antioxidative Enzymes

doi: 10.1681/ASN.2010040367

Figure Lengend Snippet: Expressions of miR-335 and miR-34a in aging mesangial cells, endothelial cells, tubular epithelial cells and interstitial fibroblasts were significantly upregulated and expressions of SOD2 and Txnrd2 in aging mesangial cells were downregulated. (A) Expression levels of miR-335 and miR-34a in aging renal mesangial cells were detected by qRT-PCR. n = 5 per miRNA. *P < 0.01 versus young. (B) Levels of SOD2 and Txnrd2 proteins were analyzed by Western blot in aging renal mesangial cells. The graph is representative of three separate experiments. (C) Expression levels of miR-335 and miR-34a were detected by qRT-PCR in the aging glomerular epithelial cells, endothelial cells, tubular epithelial cells, and fibroblasts. n = 5 per miRNA. *P < 0.01 versus young, #P < 0.05 versus young.

Article Snippet: LC Sciences (Houston, TX) provided the miRNA microarray chip (μParaflo microfluidic chip).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Journal of the American Society of Nephrology : JASN

Article Title: miR-335 and miR-34a Promote Renal Senescence by Suppressing Mitochondrial Antioxidative Enzymes

doi: 10.1681/ASN.2010040367

Figure Lengend Snippet: (A) miR-335 and miR-34a mimics inhibit SOD2 and Txnrd2 expressions in young mesangial cells, and (B) antisense miR-335 and miR-34a increase SOD2 and Txnrd2 expressions in aging mesangial cells. Co, miRNA control; CA, antisense miRNA control; miR335 and miR34a, miR-335 and miR-34a mimics, respectively; AmiR335 and AmiR34a, antisense miR-335 and miR-34a inhibitors, respectively. (C) Analysis of expression levels of SOD2 and Txnrd2 by Western blot in aged renal tissues. The graph is representative of three separate experiments.

Article Snippet: LC Sciences (Houston, TX) provided the miRNA microarray chip (μParaflo microfluidic chip).

Techniques: Control, Expressing, Western Blot

Journal: Journal of the American Society of Nephrology : JASN

Article Title: miR-335 and miR-34a Promote Renal Senescence by Suppressing Mitochondrial Antioxidative Enzymes

doi: 10.1681/ASN.2010040367

Figure Lengend Snippet: miR-335 and miR-34a mimics induce premature senescent phenotypes in young mesangial cells, and antisense miR-335 and miR-34a relieve senescent phenotypes in old mesangial cells. (A) SA-β-gal staining in young mesangial cells transfected with miR-335 and miR-34a mimics. Blue precipitation in the cytoplasm was observed in the senescent cells. (B) Analysis of SAHF formation in young mesangial cells transfected with miR-335 and miR-34a mimics. The cells were stained with DAPI, and heterochromatin foci were observed in the senescent cells. (C) SA-β-gal staining in aging mesangial cells transfected with antisense miR-335 and miR-34a inhibitors. (D) Analysis of SAHF formation in aging mesangial cells transfected with antisense miR-335 and miR-34a inhibitors. Co, miRNA control; CA, antisense miRNA control. The above results of SA-β-gal staining and SAHF analysis are representative images of three experiments.

Article Snippet: LC Sciences (Houston, TX) provided the miRNA microarray chip (μParaflo microfluidic chip).

Techniques: Staining, Transfection, Control

Journal: Leukemia

Article Title: Regulation of PI3K signaling in T cell acute lymphoblastic leukemia: a novel PTEN/Ikaros/miR-26b mechanism reveals a critical targetable role for PIK3CD

doi: 10.1038/leu.2017.80

Figure Lengend Snippet: A: miRNA profile of Array data. KO576, KO577, KO578 and KO579 are Pten -knockout mouse T-ALL samples. WT580 and WT581 are wild-type mouse thymocytes. B: Decreased miR-26b expression level in mouse Pten deficient T-ALL cell lines (LPN248, LPN236 and LPN228) compared with mouse Ink4a/Arf knock-out T-ALL cell line (LPN211) and mouse wild type thymocytes (P<0.001). C. Decreased miR-26b in human T-ALL cell lines, CCRF-CEM, SUPT1, LOUCY, KOPT-K1, JURKAT and MOLT4 compared with postnatal normal human thymus (P<0.001). D: Decreased expression level of miR-26b in human primary T-ALL samples (P<0.001). E. Correlation of miR-26b expression level with PTEN level in human primary T-ALL samples (r=0.3987, P=0.039).

Article Snippet: Hybridization of biotin-labeled cDNA was carried out on a miRNA microarray chip (MD Anderson miRNA expression Bioarray Version 5), which contains 2300 miRNA probes, including 1400 human and 900 mouse miRNA genes, in duplicate.

Techniques: Knock-Out, Expressing

Journal: British Journal of Cancer

Article Title: Dysregulation of miR-106a and miR-591 confers paclitaxel resistance to ovarian cancer

doi: 10.1038/bjc.2013.305

Figure Lengend Snippet: Significantly altered miRNAs by microRNA microarray in PTX-resistant SKpac cells compared with parent SKOV3 cells

Article Snippet: For the selection of the candidate miRNAs, the miRNA expression profiles for four PTX-resistant SKpac cell lines (SKpac-8, -10, -11, and -12) and the parent ovarian cancer cell line (SKOV3) were evaluated and compared using a bead-based miRNA microarray (Human-miRNA-V1 Bead Chips; Illumina, San Diego, CA, USA) containing 739 human precursor and mature miRNA oligonucleotide probes.

Techniques: Microarray

Journal: British Journal of Cancer

Article Title: Dysregulation of miR-106a and miR-591 confers paclitaxel resistance to ovarian cancer

doi: 10.1038/bjc.2013.305

Figure Lengend Snippet: ( A ) Hierarchical clustering of miRNA expression profiles by miRNA microarray. Unsupervised hierarchical clustering analysis of miRNAs that exhibited a statistically significant ( P <0.05) increase or decrease in PTX-resistant SKpac cells compared with PTX-sensitive SKOV3 cells. The level of miRNA expression is color-coded. ( B ) Validation by qRT–PCR of candidate miRNAs that showed significantly different expression by miRNA microarray (see A ). The bar graph shows the expression of each microRNA in SKpac cells (SKpac-8, -10, -11, -12, -16, and -17) relative to the average expression level in SKOV3 cells (* P <0.05). ( C ) Expression of miR-106a by qRT–PCR in chemosensitive and chemoresistant ovarian cancer tissues. Relative expression levels are shown normalised to benign tumours. The inner bar in each graph represents the mean value. The expression level in the chemoresistant ovarian cancers was significantly higher (3.2-fold than that in benign tissue) than in the chemosensitive cancers (1.12-fold than that in benign tissue; P =0.032). ( D ) Kaplan–Meier OS curve. The higher expression group of miR-106a (more than two-fold than that of benign tumour) demonstrated a significantly worse OS ( P =0.007) in patients with ovarian serous carcinomas.

Article Snippet: For the selection of the candidate miRNAs, the miRNA expression profiles for four PTX-resistant SKpac cell lines (SKpac-8, -10, -11, and -12) and the parent ovarian cancer cell line (SKOV3) were evaluated and compared using a bead-based miRNA microarray (Human-miRNA-V1 Bead Chips; Illumina, San Diego, CA, USA) containing 739 human precursor and mature miRNA oligonucleotide probes.

Techniques: Expressing, Microarray, Biomarker Discovery, Quantitative RT-PCR

Journal: British Journal of Cancer

Article Title: Dysregulation of miR-106a and miR-591 confers paclitaxel resistance to ovarian cancer

doi: 10.1038/bjc.2013.305

Figure Lengend Snippet: TUNEL assay in SKpac cells after transfection of anti-miR-106a or pre-miR-591. ( A ) Representative graphs of TUNEL assay. Transfection anti-miR-106a or pre-miR-591 markedly increases apoptosis of PTX-resistant SKpac cells following 80 nℳ PTX treatment. ( B ) The graph represents the mean increase in apoptosis after transfection of each miRNA following PTX treatment compared with PTX-treated, control miRNA-transfected cells (* P <0.05). ( C ) The graph represents the mean decrease in PTX (20 nℳ)-induced apoptosis in chemosensitive parental SKOV3 cells after transfection with pre-miR-106a, or anti-miR-591. The graph represents the mean±standard error of triplicate experiments.

Article Snippet: For the selection of the candidate miRNAs, the miRNA expression profiles for four PTX-resistant SKpac cell lines (SKpac-8, -10, -11, and -12) and the parent ovarian cancer cell line (SKOV3) were evaluated and compared using a bead-based miRNA microarray (Human-miRNA-V1 Bead Chips; Illumina, San Diego, CA, USA) containing 739 human precursor and mature miRNA oligonucleotide probes.

Techniques: TUNEL Assay, Transfection, Control

Journal: British Journal of Cancer

Article Title: Dysregulation of miR-106a and miR-591 confers paclitaxel resistance to ovarian cancer

doi: 10.1038/bjc.2013.305

Figure Lengend Snippet: Cell migration and colony-forming assay. ( A ) Cell migration across collagen-coated wells was assessed using the Oris Cell Migration Assay kit. Migration of PTX-treated (80 nℳ) SKpac cells transfected with pre-miR-591 or anti-miR-106a was markedly decreased compared with that of contol mRNA-transfected cells. The graph represents the mean±standard error of triplicate experiments (* P <0.05). ( B ) The mean number of colonies formed by SKpac cells transfected with anti-miR-106a or pre-miR-591 decreased compared with PTX-treated, control miRNA-transfected cells. The graph represents the mean±standard error of triplicate experiments (* P <0.05). ( C ) Colony formation by PTX-treated SKOV3 cells transfected with pre-miR-106a or anti-miR-591 was markedly increased (4–4.5-fold) compared with PTX-treated, control miRNA-transfected cells (* P <0.05).

Article Snippet: For the selection of the candidate miRNAs, the miRNA expression profiles for four PTX-resistant SKpac cell lines (SKpac-8, -10, -11, and -12) and the parent ovarian cancer cell line (SKOV3) were evaluated and compared using a bead-based miRNA microarray (Human-miRNA-V1 Bead Chips; Illumina, San Diego, CA, USA) containing 739 human precursor and mature miRNA oligonucleotide probes.

Techniques: Migration, Cell Migration Assay, Transfection, Control

Journal: British Journal of Cancer

Article Title: Dysregulation of miR-106a and miR-591 confers paclitaxel resistance to ovarian cancer

doi: 10.1038/bjc.2013.305

Figure Lengend Snippet: Luciferase assays for putative target genes of miR-106a and miR-591. The wild-type or mutated binding sites were cloned separately into the pGL3-control vector. The pGL3-control (100 ng) and pRL-TK plasmids (5 ng) were transfected, and synthetic pre-miR-591 or pre-miR-106a was added into SKpac cells. All experiments were performed in triplicate and normalised to Renilla luciferase activity. ( A ) The luciferase activity of reporter construct containing the wild-type ZEB1 3′-UTR was repressed by pre-miR-591 (50%), whereas this miRNA had no effect on the luciferase activity of reporter constructs containing mutant ZEB1 3′-UTR. ( B ) The luciferase activity of reporter construct containing the wild-type caspase-7 3′-UTR was repressed by pre-miR-106a (42%), whereas this miRNA had no effect on the luciferase activity with mutant caspase7 3′-UTR. ( C ) The luciferase activity of reporter construct containing the wild-type BCL10 3′-UTR was repressed by pre-miR-106a (40%), whereas this miRNA had no effect on the luciferase activity with mutant BCL10 3′-UTR. MT, mutant type; WT, wild type.

Article Snippet: For the selection of the candidate miRNAs, the miRNA expression profiles for four PTX-resistant SKpac cell lines (SKpac-8, -10, -11, and -12) and the parent ovarian cancer cell line (SKOV3) were evaluated and compared using a bead-based miRNA microarray (Human-miRNA-V1 Bead Chips; Illumina, San Diego, CA, USA) containing 739 human precursor and mature miRNA oligonucleotide probes.

Techniques: Luciferase, Binding Assay, Clone Assay, Control, Plasmid Preparation, Transfection, Activity Assay, Construct, Mutagenesis