Journal: Journal of Cellular and Molecular Medicine

Article Title: Screening differential circular RNA expression profiles reveals the regulatory role of circMARS in anti‐tuberculosis drug‐induced liver injury

doi: 10.1111/jcmm.17157

Figure Lengend Snippet: Study flow chart. First, the difference of circRNA expression profiles between anti‐tuberculosis drug‐induced liver injury (ADLI) and non‐ADLI patients were explored. Then, a differentially expressed circRNA was selected for verification in a cohort of 300 patients. Finally, the function of this circRNA was verified in a self‐controlled cohort of 35 patients based on the results of the experimental study

Article Snippet: The circRNAs in the discovery cohort were profiled using the CapitalBio Technology CircRNA Human Gene Expression Microarray v2.0 (CapitalBio Technology, Beijing, China).

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Screening differential circular RNA expression profiles reveals the regulatory role of circMARS in anti‐tuberculosis drug‐induced liver injury

doi: 10.1111/jcmm.17157

Figure Lengend Snippet: Identification of circRNA expression profiles in ADLI. (A) The scatter plots of circRNAs expression variations in the ADLI patients and in vitro assays. The red and green points in the plot indicate the upregulated and downregulated circRNAs. (B) Venn diagrams of the co‐expression circRNAs in the serum and cells. Two drugs: cells with Isoniazid (INH) + Rifampicin (RFP); three drugs: cells with INH + RFP + Pyrazinamide (PZA)

Article Snippet: The circRNAs in the discovery cohort were profiled using the CapitalBio Technology CircRNA Human Gene Expression Microarray v2.0 (CapitalBio Technology, Beijing, China).

Techniques: Expressing, In Vitro

Journal: Aging (Albany NY)

Article Title: Tumor-derived exosomal circRNA_102481 contributes to EGFR-TKIs resistance via the miR-30a-5p/ROR1 axis in non-small cell lung cancer

doi: 10.18632/aging.203011

Figure Lengend Snippet: The RT-qPCR primers sequences.

Article Snippet: The exosomes RNA of five pair’s samples was extracted using RNeasy Mini Kit (Qiagen GmbH) and the circRNA profile was measured using Arraystar high-throughput human circRNA microarray V2.0 (8x15 K; Arraystar, Inc.).

Techniques:

Journal: Aging (Albany NY)

Article Title: Tumor-derived exosomal circRNA_102481 contributes to EGFR-TKIs resistance via the miR-30a-5p/ROR1 axis in non-small cell lung cancer

doi: 10.18632/aging.203011

Figure Lengend Snippet: circRNA_102481 is significantly up-regulated in serum exosomes of patients with EGFR-TKIs resistance. ( A ) Hierarchical clustering analysis showed the top 10 most increased and increased circRNAs in serum exosomes. ( B ) RT-qPCR validation.

Article Snippet: The exosomes RNA of five pair’s samples was extracted using RNeasy Mini Kit (Qiagen GmbH) and the circRNA profile was measured using Arraystar high-throughput human circRNA microarray V2.0 (8x15 K; Arraystar, Inc.).

Techniques: Quantitative RT-PCR, Biomarker Discovery

Journal: Aging (Albany NY)

Article Title: Tumor-derived exosomal circRNA_102481 contributes to EGFR-TKIs resistance via the miR-30a-5p/ROR1 axis in non-small cell lung cancer

doi: 10.18632/aging.203011

Figure Lengend Snippet: circRNA_102481 is mainly secreted via exosomes. ( A , B ) circRNA_102481 was unregulated in EGFR-TKIs resistant cells (PC9/GR and HCC827/ER), ** p < 0.01 versus matched sensitive cells (PC9 and HCC827). ( C , D ) circRNA_102481 was significantly up-regulated in exosomes of EGFR-TKIs resistant cells (PC9/GR and HCC827/ER), ** p < 0.01 versus matched sensitive cells (PC9 and HCC827).

Article Snippet: The exosomes RNA of five pair’s samples was extracted using RNeasy Mini Kit (Qiagen GmbH) and the circRNA profile was measured using Arraystar high-throughput human circRNA microarray V2.0 (8x15 K; Arraystar, Inc.).

Techniques:

Journal: Aging (Albany NY)

Article Title: Tumor-derived exosomal circRNA_102481 contributes to EGFR-TKIs resistance via the miR-30a-5p/ROR1 axis in non-small cell lung cancer

doi: 10.18632/aging.203011

Figure Lengend Snippet: Exosomal circRNA_102481 derived from EGFR-TKIs resistance cells. ( A ) circRNA_102481 expression levels in exosomes were almost equal to those in CCM, but, the levels of circRNA_102481 expression in EGFR-TKIs-resistant cells were significantly higher than in CCM and exosomes, # p > 0.05, * p < 0.05. ( B ) RT-qPCR was performed to detect the expression level of circRNA_102481 after treatment with 1 μg/ml RNase alone or combined with 0.1% Triton X-100 for 30 min, # p > 0.05, * p < 0.05 compared to the PBS control. ( C ) Transfection efficiency of circRNA_102481 was verified by a confocal laser scanning microscope and RT-qPCR assay. ** P < 0.01 versus si-NC group cells. ( D ) circRNA_102481 was significantly down- regulated in exosomes of si-circRNA_ 102481 cells. * p <0.05, ** p <0.01 versus si-NC group exosomes. CCM: cell culture medium.

Article Snippet: The exosomes RNA of five pair’s samples was extracted using RNeasy Mini Kit (Qiagen GmbH) and the circRNA profile was measured using Arraystar high-throughput human circRNA microarray V2.0 (8x15 K; Arraystar, Inc.).

Techniques: Derivative Assay, Expressing, Quantitative RT-PCR, Control, Transfection, Laser-Scanning Microscopy, Cell Culture

Journal: Aging (Albany NY)

Article Title: Tumor-derived exosomal circRNA_102481 contributes to EGFR-TKIs resistance via the miR-30a-5p/ROR1 axis in non-small cell lung cancer

doi: 10.18632/aging.203011

Figure Lengend Snippet: Functional validation assay of circRNA_102481 on EGFR-TKIs resistant cells in vitro (n=3). ( A , B ) MTT assay showed the effects of circRNA_102481 on EGFR-TKIs resistant cell proliferation. ( C , D ) Annexin V/PI assay showed the effects of circRNA_102481 on EGFR-TKIs resistant cell apoptosis. si-circRNA_102481: siRNA targeting circRNA_102481. ** p <0.01 versus si-NC group.

Article Snippet: The exosomes RNA of five pair’s samples was extracted using RNeasy Mini Kit (Qiagen GmbH) and the circRNA profile was measured using Arraystar high-throughput human circRNA microarray V2.0 (8x15 K; Arraystar, Inc.).

Techniques: Functional Assay, Biomarker Discovery, In Vitro, MTT Assay

Journal: Aging (Albany NY)

Article Title: Tumor-derived exosomal circRNA_102481 contributes to EGFR-TKIs resistance via the miR-30a-5p/ROR1 axis in non-small cell lung cancer

doi: 10.18632/aging.203011

Figure Lengend Snippet: Functional validation assay of exosomes circRNA_102481 on EGFR-TKIs resistant cells in vitro (n=3). ( A , B ) MTT assay showed the effects of exosomes circRNA_102481 on EGFR-TKIs resistant cell proliferation. ( C , D ) Annexin V/PI assay showed the effects of exosomes circRNA_102481 on EGFR-TKIs resistant cell apoptosis. si-circRNA_102481: siRNA targeting circRNA_102481. * p <0.05 versus si-NC group.

Article Snippet: The exosomes RNA of five pair’s samples was extracted using RNeasy Mini Kit (Qiagen GmbH) and the circRNA profile was measured using Arraystar high-throughput human circRNA microarray V2.0 (8x15 K; Arraystar, Inc.).

Techniques: Functional Assay, Biomarker Discovery, In Vitro, MTT Assay

Journal: Aging (Albany NY)

Article Title: Tumor-derived exosomal circRNA_102481 contributes to EGFR-TKIs resistance via the miR-30a-5p/ROR1 axis in non-small cell lung cancer

doi: 10.18632/aging.203011

Figure Lengend Snippet: Functional validation assay of exosomes circRNA_102481 on EGFR-TKIs sensitive cells in vitro (n=3). ( A , B ) MTT assay showed the effects of circRNA_102481 on EGFR-TKIs sensitive cell proliferation. ( C , D ) Annexin V/PI assay showed the effects of circRNA_102481 on EGFR-TKIs sensitive cell apoptosis. circRNA_100859 vector: circRNA_100859 overexpression vector. * p <0.05 versus blank vector group.

Article Snippet: The exosomes RNA of five pair’s samples was extracted using RNeasy Mini Kit (Qiagen GmbH) and the circRNA profile was measured using Arraystar high-throughput human circRNA microarray V2.0 (8x15 K; Arraystar, Inc.).

Techniques: Functional Assay, Biomarker Discovery, In Vitro, MTT Assay, Plasmid Preparation, Over Expression

Journal: Aging (Albany NY)

Article Title: Tumor-derived exosomal circRNA_102481 contributes to EGFR-TKIs resistance via the miR-30a-5p/ROR1 axis in non-small cell lung cancer

doi: 10.18632/aging.203011

Figure Lengend Snippet: circRNA_102481 serves as a miR-30a-5p sponge to regulate ROR1 expression. The detailed potential binding sequences of circRNA_102481 to miR-30a-5p ( A ) and ROR1 to miR-30a-5p ( D ). miR-30a-5p of PC9/GR and HCC827/ER cells was abundantly pulled down by circRNA_102481 ( B ) or ROR1 ( E ). Dual-luciferase reporter assay confirmed that miR-30a-5p competitively targeted circRNA_102481( C ), and that ROR1 is a direct target of miR-30a-5p ( F ). # p > 0.05, * p < 0.05, ** p <0.01.

Article Snippet: The exosomes RNA of five pair’s samples was extracted using RNeasy Mini Kit (Qiagen GmbH) and the circRNA profile was measured using Arraystar high-throughput human circRNA microarray V2.0 (8x15 K; Arraystar, Inc.).

Techniques: Expressing, Binding Assay, Luciferase, Reporter Assay

Journal: Aging (Albany NY)

Article Title: Tumor-derived exosomal circRNA_102481 contributes to EGFR-TKIs resistance via the miR-30a-5p/ROR1 axis in non-small cell lung cancer

doi: 10.18632/aging.203011

Figure Lengend Snippet: Effects of exosomes on circRNA_102481-miR-30a-5p-ROR1 Axis. ( A , B ) RT-qPCR assay showed miR-30a-5p expression in PC9/GR or HCC827/ER cells of si-NC and si-circRNA_102481 groups. ( C ) Transfection efficiency of miR-30a-5p mimic and miR-30a-5p inhibitor was verified by RT-qPCR assay. ** p <0.01 versus miR-30a-5p NC group cells. ( D , E ) qRT-PCR demonstrated that ROR1 mRNA expression were significantly decreased in miR-30a-5p mimic group, and increased in miR-30a-5p inhibitor group. * p <0.05, ** p <0.01 versus miR-30a-5p NC group.

Article Snippet: The exosomes RNA of five pair’s samples was extracted using RNeasy Mini Kit (Qiagen GmbH) and the circRNA profile was measured using Arraystar high-throughput human circRNA microarray V2.0 (8x15 K; Arraystar, Inc.).

Techniques: Quantitative RT-PCR, Expressing, Transfection

Journal: Aging (Albany NY)

Article Title: Tumor-derived exosomal circRNA_102481 contributes to EGFR-TKIs resistance via the miR-30a-5p/ROR1 axis in non-small cell lung cancer

doi: 10.18632/aging.203011

Figure Lengend Snippet: Effects of exosomes on circRNA_102481-miR-30a-5p-ROR1 Axis. ( A , B ) Relative ROR1 mRNA and protein were analyzed in PC9/GR and HCC827/ER cells the cells were co-cultured with exosomes of the si-NC, si-circRNA_102481, and miR-30a-5p NC, miR-30a-5p inhibitor using RT-qPCR and western blot assay, respectively. ( C ) IC50 of gefitinib or erlotinib the cells were co-cultured with exosomes of the si-NC, si-circRNA_102481, and miR-30a-5p NC, miR-30a-5p inhibitor was detected by MTT assay. * p < 0.05, ** p <0.01.

Article Snippet: The exosomes RNA of five pair’s samples was extracted using RNeasy Mini Kit (Qiagen GmbH) and the circRNA profile was measured using Arraystar high-throughput human circRNA microarray V2.0 (8x15 K; Arraystar, Inc.).

Techniques: Cell Culture, Quantitative RT-PCR, Western Blot, MTT Assay

Journal: Aging (Albany NY)

Article Title: Tumor-derived exosomal circRNA_102481 contributes to EGFR-TKIs resistance via the miR-30a-5p/ROR1 axis in non-small cell lung cancer

doi: 10.18632/aging.203011

Figure Lengend Snippet: Large sample validation and correlation analysis. ( A – C ) RT-qPCR analysis of exosomes circRNA_102481, exosomes miR-30a-5p, exosomes ROR1 mRNA before and after EGFR-TKIs resistance in 58 NSCLC patients. ( D , E ) Pearson’s correlation analysis of between exosomes circRNA_102481 and exosomes miR-30a-5p, between exosomes miR-30a-5p and exosomes ROR1 mRNA.

Article Snippet: The exosomes RNA of five pair’s samples was extracted using RNeasy Mini Kit (Qiagen GmbH) and the circRNA profile was measured using Arraystar high-throughput human circRNA microarray V2.0 (8x15 K; Arraystar, Inc.).

Techniques: Biomarker Discovery, Quantitative RT-PCR

Journal: Aging (Albany NY)

Article Title: Tumor-derived exosomal circRNA_102481 contributes to EGFR-TKIs resistance via the miR-30a-5p/ROR1 axis in non-small cell lung cancer

doi: 10.18632/aging.203011

Figure Lengend Snippet: Relationship between exosomes circRNA_102481-miR-30a-5p-ROR1 axis with clinicopathological variables. ( A ) exosomes circRNA_102481, ( B ) exosomes miR-30a-5p, ( C ) exosomes ROR1 mRNA.

Article Snippet: The exosomes RNA of five pair’s samples was extracted using RNeasy Mini Kit (Qiagen GmbH) and the circRNA profile was measured using Arraystar high-throughput human circRNA microarray V2.0 (8x15 K; Arraystar, Inc.).

Techniques:

Journal: Aging (Albany NY)

Article Title: Tumor-derived exosomal circRNA_102481 contributes to EGFR-TKIs resistance via the miR-30a-5p/ROR1 axis in non-small cell lung cancer

doi: 10.18632/aging.203011

Figure Lengend Snippet: Kaplan-Meier PFS curve stratified by exosomes circRNA_102481- miR-30a-5p-ROR1 axis expression. PFS duration between higher and lower ( A ) exosomes circRNA_102481, ( B ) exosomes miR-30a-5p, ( C ) exosomes ROR1 mRNA.

Article Snippet: The exosomes RNA of five pair’s samples was extracted using RNeasy Mini Kit (Qiagen GmbH) and the circRNA profile was measured using Arraystar high-throughput human circRNA microarray V2.0 (8x15 K; Arraystar, Inc.).

Techniques: Expressing

Journal: Aging (Albany NY)

Article Title: Tumor-derived exosomal circRNA_102481 contributes to EGFR-TKIs resistance via the miR-30a-5p/ROR1 axis in non-small cell lung cancer

doi: 10.18632/aging.203011

Figure Lengend Snippet: Kaplan-Meier OS curve stratified by exosomes circRNA_102481- miR-30a-5p-ROR1 axis expression. OS duration between higher and lower ( A ) exosomes circRNA_102481, ( B ) exosomes miR-30a-5p, ( C ) exosomes ROR1 mRNA.

Article Snippet: The exosomes RNA of five pair’s samples was extracted using RNeasy Mini Kit (Qiagen GmbH) and the circRNA profile was measured using Arraystar high-throughput human circRNA microarray V2.0 (8x15 K; Arraystar, Inc.).

Techniques: Expressing

Journal: Clinical and Translational Medicine

Article Title: Androgen receptor decreases the renal cell carcinoma bone metastases via suppressing the osteolytic formation through altering a novel circEXOC7 regulatory axis

doi: 10.1002/ctm2.353

Figure Lengend Snippet: CircRNA microarray and screening process. Results from circRNA microarray and bioinformatics analysis from five paired human RCC vs. RBM tissues were summarized in (A‐D). (A) Box plot demonstrated no significant differences in the distribution of intensity across 10 clinical samples. (B) Scatter plot displayed circRNAs expression correlation between RCC and RBM group. (C) Heat map generated from circRNAs microarray data reflecting circRNAs expression values in RCC and RBM groups. (D) Volcano plot showing DERs data. The red points indicate points‐of‐interest that display both large magnitude fold‐changes (x‐axis) and high statistical significance (‐log10 of p ‐value, y‐axis). The green line shows points above the line having p < 0.05 and points below the line having p > 0.05. This plot is colored such that those points having a fold‐change less than 2 (log2 = 1) are shown in gray. (E) The qRT‐PCR was performed to show 32 selected candidate circRNAs expression levels in cells. (F) The qRT‐PCR was performed on SW839 cells to show effects of manipulating efficiency of selected candidate circRNAs, ciR4#, ciR6#, ciR19#, and ciR26#. (G‐J) Interruption assays revealed that suppressing (G) ciR‐4# (hsa_circ_0001275), (H) ciR‐6# (hsa_circRNA_405974), expression cannot reverse/block the knocked‐down AR‐increased osteolytic formation in SW839 cells (scale bars, 200 μm), increasing (I) ciR‐19# (hsa_circ_0091743) expression, and (J) increasing ciR‐26# hsa_circ_0075303 expression cannot reverse/block the AR‐suppressed osteolytic formation in SW839 cell line (scale bars, 200 μm). For (E‐J), quantitation is at the right, and data are presented as Mean ± SD, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Abbreviation: N.S., not significant

Article Snippet: In the current study, five human RCC and five RCC bone metastasis tissues were deeply sequenced using Arraystar human circRNA V2.0 microarray.

Techniques: Microarray, Expressing, Generated, Quantitative RT-PCR, Blocking Assay, Quantitation Assay

Journal: Clinical and Translational Medicine

Article Title: Androgen receptor decreases the renal cell carcinoma bone metastases via suppressing the osteolytic formation through altering a novel circEXOC7 regulatory axis

doi: 10.1002/ctm2.353

Figure Lengend Snippet: The expression of circEXOC7 is regulated by DHX9. (A) The qRT‐PCR was performed on SW839 and OS‐RC‐2 cell lines to show correlation between AR and four genes related to circRNA biogenesis. Among them, ADAR2 and DHX9 have a positive correlation with AR mRNA expression. (B and C) The qRT‐qPCR was performed to show (B) knocking down DHX9, and not (C) ADAR2 gene, led to increased circEXOC7 expression in SW839 cells. (D) WB was performed on SW839 and OS‐RC‐2 cells to show that modulated AR expression also altered DHX9 expression. (E) Predicted AR binding sites on the DHX9 promoter region using website http://jaspar.genereg.net/ . (F) ChIP assay showing AR could bind to the No. 2 potential ARE on the DHX9 5′‐promoter region. (G) The wild type and mutant pGL3‐DHX9 promoter reporter constructs. (H and I) Luciferase activity after transfection of wild type or mutant DHX9 promoter reporter constructs in OS‐RC‐2 cells (H) transfected with AR‐cDNA or pWPI and SW839 cells (I) transfected with AR‐shRNA or pLKO. For (B and C), (H and I), data are presented as Mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001. Abbreviation: N.S., not significant

Article Snippet: In the current study, five human RCC and five RCC bone metastasis tissues were deeply sequenced using Arraystar human circRNA V2.0 microarray.

Techniques: Expressing, Quantitative RT-PCR, Binding Assay, Mutagenesis, Construct, Luciferase, Activity Assay, Transfection, shRNA