Journal: American Journal of Cancer Research

Article Title: LINC01354 enhances the metastatic ability of gastric cancer cells by adjusting miR-153-5p/CADM2 expression

doi:

Figure Lengend Snippet: Higher expression of LINC01354 and lower expression of miR-153-5p was observed in GC tissues and cell lines. A. LINC01354 expression in GC cancer tissue (Ct) and paracancerous tissue (Pt) was detected by qRT-PCR. B. The predicted targeting sites of LINC01354 binding to miR-153-5p were shown in database LncBase v.2. C. miR-153-5p expression was detected by qRT-PCR. D. Correlation between the expression level of LINC01354 and miR-153-5p. E, F. qRT-PCR was performed to determine the expression of LINC01354 and miR-153-5p in GC cell lines. G. The subcellular location of LINC01354 in HGC-27 and NCI-N87 cells was examined by fluorescence in situ hybridization assay. **P < 0.01, ***P < 0.001.

Article Snippet: Cyanine 3-labeled fluorescent probes specifically targeting LINC01354 were synthesized by RiBobio Inc. (Guangzhou, China) to explore the subcellular location of LINC01354 in GC cells.

Techniques: Expressing, Quantitative RT-PCR, Binding Assay, Fluorescence, In Situ Hybridization

Journal: American Journal of Cancer Research

Article Title: LINC01354 enhances the metastatic ability of gastric cancer cells by adjusting miR-153-5p/CADM2 expression

doi:

Figure Lengend Snippet: Abnormally high expression of CADM2 in GC tissues. A. Wild-type LINC01354 site (Site1-4 WT) or mutant-type LINC01354 (Site1-4 MUT) reporters and NC mimic or miR-153-5p were co-transfected into HEK-293T cells. Firefly/Renilla luciferase activity was measured. B. The binding site of miR-153-5p to CADM2 3’UTR was shown in the database TargetScanHuman 7.2. C. A dual-luciferase reporter assay was used to assess the relation between miR-153-5p and CADM2. D, E. CADM2 expression and its relation to the survival of GC patients were shown in the TCGA Database. F. CADM2 expression in GC cancer tissue (Ct) and paracancerous tissue (Pt) was measured by qRT-PCR. G. Association between the levels of LINC01354 and CADM2 mRNA. H. Association between the levels of miR-153-5p and CADM2 mRNA. ***P < 0.001.

Article Snippet: Cyanine 3-labeled fluorescent probes specifically targeting LINC01354 were synthesized by RiBobio Inc. (Guangzhou, China) to explore the subcellular location of LINC01354 in GC cells.

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

Journal: American Journal of Cancer Research

Article Title: LINC01354 enhances the metastatic ability of gastric cancer cells by adjusting miR-153-5p/CADM2 expression

doi:

Figure Lengend Snippet: LINC01354 knockdown inhibited EMT progress by decreasing CADM2 expression. LINC01354 knockdown was performed in NCI-N87 and HGC-27 cells. A. Levels of LINC01354, miR-153-5p, and CADM2 mRNA were measured by qRT-PCR. B. Proteins of CADM2, E-cadherin, N-cadherin, and Vimentin were determined by western blotting. C, D. Proteins of CADM2, E-cadherin, and N-cadherin were assessed by immunofluorescence assay. E. Cell activity was assessed by a CCK8 assay. F. A wound-healing assay was performed to evaluate cell migration. G, H. Cell migration and invasion were detected by using a Transwell chamber. ***P < 0.001.

Article Snippet: Cyanine 3-labeled fluorescent probes specifically targeting LINC01354 were synthesized by RiBobio Inc. (Guangzhou, China) to explore the subcellular location of LINC01354 in GC cells.

Techniques: Knockdown, Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Activity Assay, CCK-8 Assay, Wound Healing Assay, Migration

Journal: American Journal of Cancer Research

Article Title: LINC01354 enhances the metastatic ability of gastric cancer cells by adjusting miR-153-5p/CADM2 expression

doi:

Figure Lengend Snippet: miR-153-5p overexpression attenuated the influence of LINC01354 on GC cells. LINC01354 and miR-153-5p were co-expressed in MKN-45 and SNU-1 cells. A. The levels of LINC01354, miR-153-5p, and CADM2 mRNA were measured via qRT-PCR. B. Proteins of CADM2, E-cadherin, N-cadherin, and Vimentin were determined by western blotting. C, D. Proteins of CADM2, E-cadherin, and N-cadherin were explored by immunofluorescence assay. E. A wound-healing assay was performed to assess cell migration. F, G. Cell migration and invasion were detected by using a Transwell chamber. ***P < 0.001, vs. Vector group; ###P < 0.001, vs. LINC01354 group.

Article Snippet: Cyanine 3-labeled fluorescent probes specifically targeting LINC01354 were synthesized by RiBobio Inc. (Guangzhou, China) to explore the subcellular location of LINC01354 in GC cells.

Techniques: Over Expression, Quantitative RT-PCR, Western Blot, Immunofluorescence, Wound Healing Assay, Migration, Plasmid Preparation

Journal: American Journal of Cancer Research

Article Title: LINC01354 enhances the metastatic ability of gastric cancer cells by adjusting miR-153-5p/CADM2 expression

doi:

Figure Lengend Snippet: CADM2 knockdown antagonized the effects of LINC01354 overexpression. LINC01354 overexpression plasmid and siRNA-targeted CADM2 were co-transfected in MKN-45 and SNU-1 cells. A. Proteins of CADM2, E-cadherin, N-cadherin, and Vimentin were determined through western blotting. B. A wound-healing assay was performed to assess cell migration. C, D. Cell migration and invasion were detected by a Transwell chamber. ***P < 0.001, vs. Vector+siNC group; ###P < 0.001, vs. LINC01354+siNC group.

Article Snippet: Cyanine 3-labeled fluorescent probes specifically targeting LINC01354 were synthesized by RiBobio Inc. (Guangzhou, China) to explore the subcellular location of LINC01354 in GC cells.

Techniques: Knockdown, Over Expression, Plasmid Preparation, Transfection, Western Blot, Wound Healing Assay, Migration

Journal: American Journal of Cancer Research

Article Title: LINC01354 enhances the metastatic ability of gastric cancer cells by adjusting miR-153-5p/CADM2 expression

doi:

Figure Lengend Snippet: CADM2 overexpression inhibited the influence of miR-153-5p on GC cells. CADM2 and miR-153-5p were co-expressed in NCI-N87 and HGC-27 cells. A. Levels of LINC01354, miR-153-5p, and CADM2 mRNA were measured by qRT-PCR. B. Proteins of CADM2, E-cadherin, N-cadherin, and Vimentin were determined by western blotting. C, D. Proteins of CADM2, E-cadherin, and N-cadherin were evaluated by immunofluorescence assay. E. A wound-healing assay was performed to assess cell migration. F, G. Cell migration and invasion were detected by a Transwell chamber. ***P < 0.001, vs. Vector group; ###P < 0.001, vs. miR-153-5p group.

Article Snippet: Cyanine 3-labeled fluorescent probes specifically targeting LINC01354 were synthesized by RiBobio Inc. (Guangzhou, China) to explore the subcellular location of LINC01354 in GC cells.

Techniques: Over Expression, Quantitative RT-PCR, Western Blot, Immunofluorescence, Wound Healing Assay, Migration, Plasmid Preparation

Journal: Journal of Translational Medicine

Article Title: circCRKL, a circRNA derived from CRKL, regulates BCR-ABL via sponging miR-877-5p to promote chronic myeloid leukemia cell proliferation

doi: 10.1186/s12967-022-03586-2

Figure Lengend Snippet: Identifying and characterizing circCRKL in BCR-ABL + cell lines and CML samples. A Differential expression of circCRKL between CML and normal donors samples. B Relative circCRKL expression in a normal lymphoblastic cell line (TK6) and BCR-ABL − leukemia cell line (THP-1) and BCR-ABL + leukemia cell lines (K562, K562/G01, KCL22, and SupB15). C Schematic diagram of circularization of circCRKL, the back-splice site was identified using sanger sequencing. The cytoplasmic distribution of circCRKL in K562 cells ( D ) and K562/G01 cells ( E ). F FISH assay demonstrated that circCRKL was predominantly expressed in cytoplasm in K562 and K562/G01 cells (Cy3 was used for labeling circCRKL probe). G , H After treating K562 and K562/G01 cells with or without RNase R, the relative expression of circCRKL and linear CRKL was measured. I , J Relative expression of circCRKL and linear CRKL following treatment with actinomycin D in K562 and K562/G01 cells. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** < 0.0001

Article Snippet: The specific probe targeting the back-splicing site of circCRKL and NC oligo probe was coupled by biotin and purchased from RiboBio (Guangzhou, China).

Techniques: Expressing, Sequencing, Labeling

Journal: Journal of Translational Medicine

Article Title: circCRKL, a circRNA derived from CRKL, regulates BCR-ABL via sponging miR-877-5p to promote chronic myeloid leukemia cell proliferation

doi: 10.1186/s12967-022-03586-2

Figure Lengend Snippet: circCRKL promotes CML cells proliferation in vitro. The efficient and specific knockdown of circCRKL in K562 ( A ) and K562/G01 ( B ) cells. C , D CCK-8 assays were used to investigate the effect of circCRKL knockdown on growth in K562 ( C ) and K562/G01 ( D ) cells. E , F Effect of circCRKL knockdown on proliferation in K562 and K562/G01 cells was determined by colony formation assays. G , H Flow cytometry was used to analyze the effect of circCRKL suppression on the cell cycle. * p < 0.05, ** p < 0.01, and *** p < 0.001

Article Snippet: The specific probe targeting the back-splicing site of circCRKL and NC oligo probe was coupled by biotin and purchased from RiboBio (Guangzhou, China).

Techniques: In Vitro, CCK-8 Assay, Flow Cytometry

Journal: Journal of Translational Medicine

Article Title: circCRKL, a circRNA derived from CRKL, regulates BCR-ABL via sponging miR-877-5p to promote chronic myeloid leukemia cell proliferation

doi: 10.1186/s12967-022-03586-2

Figure Lengend Snippet: circCRKL enhances CML cells proliferation in vivo. A The number of WBCs was calculated. B , C The weights of liver and spleen were recorded and the images were shown. D Wright’s staining was performed to observe the leukemic cells in bone marrow, liver, and spleen, the leukemic cells were indicated by the black arrow. Scale bar, 10 μm. E The leukemic infiltration in liver and spleen was observed by H&E staining. F BCR-ABL levels in bone marrow, liver, and spleen cells were determined with immunofluorescence. Scale bar, 10 μm. * p < 0.05 and ** < 0.01

Article Snippet: The specific probe targeting the back-splicing site of circCRKL and NC oligo probe was coupled by biotin and purchased from RiboBio (Guangzhou, China).

Techniques: In Vivo, Staining, Immunofluorescence

Journal: Journal of Translational Medicine

Article Title: circCRKL, a circRNA derived from CRKL, regulates BCR-ABL via sponging miR-877-5p to promote chronic myeloid leukemia cell proliferation

doi: 10.1186/s12967-022-03586-2

Figure Lengend Snippet: BCR-ABL levels were downregulated in circCRKL silencing CML cells. Effect of circCRKL silencing on BCR-ABL protein levels was evaluated by western bolt assays ( A ) and immunofluorescence ( B ). Scale bar, 10 μm. C Effect of circCRKL knockdown on BCR-ABL mRNA levels in CML cells was determined with RT-qPCR. D The Venn diagram illustrating the overlapped miRNAs for circCRKL predicted using ENCORI, circInteractome, and circBank. E The Venn diagram showed the overlapped miRNAs between circCRKL targets and candidates for ABL obtained from ENCORI, TargetScan, and miRwalk. F The schematic diagrams of binding sites of circCRKL with miR-877-5p, and ABL 3’UTR with miR-877-5p. G , H Relative luciferase activities of the reporter plasmids were assessed in 293 T cells treated with miR-877-5p mimic or miR-NC. I Relative enrichment of circCRKL and miR-877-5p on circCRKL probe in K562/G01 cells was evaluated with RNA pulldown assay. J Relative enrichment of circCRKL and miR-877-5p on AGO2 protein in K562/G01 cells was detected by RNA immunoprecipitation. * p < 0.05 and ** < 0.01

Article Snippet: The specific probe targeting the back-splicing site of circCRKL and NC oligo probe was coupled by biotin and purchased from RiboBio (Guangzhou, China).

Techniques: Western Blot, Immunofluorescence, Quantitative RT-PCR, Binding Assay, Luciferase, Immunoprecipitation

Journal: Journal of Translational Medicine

Article Title: circCRKL, a circRNA derived from CRKL, regulates BCR-ABL via sponging miR-877-5p to promote chronic myeloid leukemia cell proliferation

doi: 10.1186/s12967-022-03586-2

Figure Lengend Snippet: circCRKL absorbs miR-877-5p to augment BCR-ABL and to accelerate CML cells proliferation. A , B CCK-8 assay was conducted to measure the cell growth co-transfected with circCRKL knockdown lentivirus and miR-877-5p inhibitor. C , D Colony formation assay was performed to determine the proliferation in CML cells co-transfected circCRKL knockdown lentivirus and miR-877-5p inhibitor. E The BCR-ABL protein levels were detected by western blot assays with co-transfecting circCRKL knockdown lentivirus and miR-877-5p inhibitor. F The BCR-ABL protein levels were determined using western blot assays with co-transfecting circCRKL knockdown lentivirus and miR-877-5p mimic. G Schematic diagram illustrates that circCRKL promotes CML cells proliferation via miR-877-5p/BCR-ABL axis. * p < 0.05, ** p < 0.01 and *** < 0.001

Article Snippet: The specific probe targeting the back-splicing site of circCRKL and NC oligo probe was coupled by biotin and purchased from RiboBio (Guangzhou, China).

Techniques: CCK-8 Assay, Transfection, Colony Assay, Western Blot