Journal: Journal of Clinical Laboratory Analysis

Article Title: The up‐regulated hsa‐circRNA9102‐5 may be a risk factor for essential hypertension

doi: 10.1002/jcla.23339

Figure Lengend Snippet: CircRNA‐miRNA‐mRNA network diagram. The red circle represents the target circRNA hsa‐circRNA9102‐5, the blue circle represents the miRNA, and the yellow circle represents the target miRNA hsa‐miR‐150‐5p. The lines represent the connection and binding of each other

Article Snippet: Quantitative PCR was performed using the GoTaqqPCR Master Mix (Promega) and the miRcute Plus miRNA SYBR Green qPCR Detection Kit (TIANGEN) on a Roche LightCycler 480 Real Time PCR instrument (Roche), by following the manufacturers' instructions.

Techniques: Binding Assay

Journal: Molecular Cancer Therapeutics

Article Title: miR200c Attenuates P-gp–Mediated MDR and Metastasis by Targeting JNK2/c-Jun Signaling Pathway in Colorectal Cancer

doi: 10.1158/1535-7163.mct-14-0167

Figure Lengend Snippet: Figure 1. miRNA expression in MDR cancer cells and primary colorectal metastasis cancer samples. A, the different levels of miRNA screened by miRNA microarray analysis were validated with qRT-PCR in four pairs MDR cell HCT8/V, HCT116/L-OHP, SGC7901/DDP, and Bel7402/Fu versus parental cells. B, mRNA levels of ABCB1 and miR200c in primary tumor samples (n ¼ 30), hepatic metastasis (n ¼ 29), and lung metastasis (n ¼ 7) were quantified by qRT-PCR. Expression levels of ABCB1 mRNA (C) and miR200c (D) in matched primary colorectal cancer (PC) and the matched liver and lung metastasis. The bold horizontal bar represents mean expression levels (, P < 0.05; , P < 0.01, t test).

Article Snippet: RNA and miRNA extraction, quantitative RT-PCR Total RNA was isolated and purified from cultured cells, plasma, and tissue samples by the RNeasy Mini Kit (Qiagen, Inc.). miRNA was prepared by the miRcute miRNA isolation Kit (Tiangen, Co, Ltd.).

Techniques: Expressing, Microarray, Quantitative RT-PCR

Journal: Molecular Cancer Therapeutics

Article Title: miR200c Attenuates P-gp–Mediated MDR and Metastasis by Targeting JNK2/c-Jun Signaling Pathway in Colorectal Cancer

doi: 10.1158/1535-7163.mct-14-0167

Figure Lengend Snippet: Figure 2. miR200c targets JNK2 30-UTR. A, the box shows a miR200c-predicted binding site, 312–334, in ABCB1 30-UTR. The sequences of ABCB1 30-UTR and its mutants used in this study are indicated. B, Luciferase reporter assay results showing the effect of miR200c on ABCB1 30-UTR. Luciferase constructs ABCB1-WT, ABCB1-MUT, or pmiR-GLO vector alone were cotransfected with TK-Renilla plasmid into HCT8/V cells with miR200c mimics/inhibitor (miR200cover/inhibitor) or miR153 mimics/inhibitor (miR153over/inhibitor). Luciferase activity was measured and normalized with Renilla luciferase values. The mean and SEs with triplicate experiments are shown. C, the box shows a miR200c-predicted binding site, 353–381, in JNK2 30-UTR. The sequences of JNK2 30-UTR and its mutants used in this study are indicated. D, Luciferase reporter assay showing the effect of miR200c on JNK2. Luciferase constructs JNK2-WT, JNK2-MUT, or pmiR-GLO vector alone were cotransfected with TK-Renilla plasmid into HCT8/V cells. Luciferase activity was measured and normalized with Renilla luciferase values. The mean and SEs from triplicate experiments are indicated. The effect of miR153, an unrelated miRNA that does not bind 30-UTR of JNK2, is indicated. , P < 0.05; , P < 0.01, JNK2-WT vs. vector. E, the effect of miR200c manipulation on protein levels of JNK2 in MDR cell HCT8/V and itsparental cells. Left, lysates from the cells treated with miR200c mimics and inhibitor were probed for JNK2. Right, quantitative analysis of the Western blot data from the left. , P < 0.05; , P < 0.01, miR200cover vs. vector.

Article Snippet: RNA and miRNA extraction, quantitative RT-PCR Total RNA was isolated and purified from cultured cells, plasma, and tissue samples by the RNeasy Mini Kit (Qiagen, Inc.). miRNA was prepared by the miRcute miRNA isolation Kit (Tiangen, Co, Ltd.).

Techniques: Binding Assay, Luciferase, Reporter Assay, Construct, Plasmid Preparation, Activity Assay, Western Blot

Journal: Molecular Cancer Therapeutics

Article Title: miR200c Attenuates P-gp–Mediated MDR and Metastasis by Targeting JNK2/c-Jun Signaling Pathway in Colorectal Cancer

doi: 10.1158/1535-7163.mct-14-0167

Figure Lengend Snippet: Figure 4. miR200c modulates MDR phenotype and cancer cell invasion/migration. A, miR200c modulates the chemosensitivity of HCT8/V cells. The cell proliferation of VCR-treated HCT8/V cells transfected with miR200c mimics, JNK2 siRNA, or PEGF-JNK2. Data are presented as mean SD of triplicate experiments. , P < 0.01 vs. vector group; DD, P < 0.01 sh- JNK2 vs. JNK2over; ##, P < 0.01 JNK2over/miR200cover vs. miR200cover. B, the cell proliferation of HCT8/V cells treated with miR200c mimics (50 nmol/L) in combination with cDDP, 5-Fu, and HTP. C, miR200c sensitizes HCT8/V cells to VCR- induced apoptosis via inhibiting JNK2. Flow cytometry analysis of apoptosis (Annexin Vþ) for HCT8/V cells transfected with miRNA or siRNA control, miR200cover, sh- JNK2, JNK2over, or PEGF-JNK2, and treated with VCR (100 mg/mL, half of the IC50 based on the data shown in Supplementary Tables S6 and S7) for 24 hours. D, top, fold changes of intracellular VCR accumulation in HCT8/V cells were measured by a validated HPLC method as described in Materials and Methods. Data, mean SD from at least three experiments. , P < 0.01 vs. vector group; DD, P < 0.01 sh-JNK2 vs. JNK2over; ##, P < 0.01 JNK2over/miR200cover

Article Snippet: RNA and miRNA extraction, quantitative RT-PCR Total RNA was isolated and purified from cultured cells, plasma, and tissue samples by the RNeasy Mini Kit (Qiagen, Inc.). miRNA was prepared by the miRcute miRNA isolation Kit (Tiangen, Co, Ltd.).

Techniques: Migration, Transfection, Plasmid Preparation, Flow Cytometry, Control

Journal: Molecular Cancer Therapeutics

Article Title: miR200c Attenuates P-gp–Mediated MDR and Metastasis by Targeting JNK2/c-Jun Signaling Pathway in Colorectal Cancer

doi: 10.1158/1535-7163.mct-14-0167

Figure Lengend Snippet: Figure 6. Expression of miR-200c, JNK2, ABCB1, and MMP-2/9 after chemotherapy. A-E, large-scale validation of miR-200c, JNK2, ABCB1, and MMP-2/9 levels in patient plasma samples. Scatter plots of plasma levels of miR-200c in healthy subjects (n = 30), before chemotherapy (n = 30), and after second (n = 30), fourth (n = 30), and sixth (n = 30) cycle of chemotherapy. Expression levels of the miRNAs (fold change at y- axis) are normalized to U6. The line represents the median value. Mann-Whitney U test was used to determine statistical significance.

Article Snippet: RNA and miRNA extraction, quantitative RT-PCR Total RNA was isolated and purified from cultured cells, plasma, and tissue samples by the RNeasy Mini Kit (Qiagen, Inc.). miRNA was prepared by the miRcute miRNA isolation Kit (Tiangen, Co, Ltd.).

Techniques: Expressing, Biomarker Discovery, Clinical Proteomics, MANN-WHITNEY

Journal: Respiratory Research

Article Title: Hypoxia-preconditioned mesenchymal stromal cell–derived exosomes attenuate LPS-induced acute lung injury and early epithelial remodeling in mice: evidence for a miR-486a-5p–Skp2–GATA4 axis

doi: 10.1186/s12931-026-03560-2

Figure Lengend Snippet: HExos alleviate EMT-like changes through miR-486a-5p ( A – C ) miRNA profiling: Heatmap/volcano plot of 38/130 miRNAs in HExos; qRT-PCR validation of top 10 (including miR-486a-5p) n = 3 ( D – E ). miR-486a-5p expression: Consistently higher in LPS+HExos n = 3 ( F – H ) Functional rescue: miR-486a-5p mimics attenuated apoptosis and modulated EMT markers (WB/RT-PCR) n = 3. Data are presented as the mean ± SD. from three independent experiments, * p < 0.05

Article Snippet: For mRNA, cDNA was synthesized with PrimeScript RT reagent (Takara, Japan) and qPCR performed using SYBR Premix Ex Taq (Takara) on QuantStudio 5 (Thermo Fisher, USA). miRNA analysis employed miRcute kits (Tiangen, China; #DP501 for isolation, #KR211 for cDNA) with LightCycler ® 96 (Roche) using SYBR Green Master Mix (#FP411).

Techniques: Quantitative RT-PCR, Biomarker Discovery, Expressing, Functional Assay, Reverse Transcription Polymerase Chain Reaction