Journal: Open Medicine

Article Title: circRNF20 aggravates the malignancy of retinoblastoma depending on the regulation of miR-132-3p/PAX6 axis

doi: 10.1515/med-2022-0483

Figure Lengend Snippet: Effects of circRNF20 knockdown on RB cell proliferation, apoptosis, migration and invasion. (a) The expression of circRNF20 in Y79 and WERI-Rb-1 cells transfected with sh-NC, sh-circRNF20#1, sh-circRNF20#2 and sh-circRNF20#3 was detected by qRT-PCR assay. (b–h) Y79 and WERI-Rb-1 cells were transfected with sh-NC or sh-circRNF20#1. (b–d) The viability, proliferation and colony formation of Y79 and WERI-Rb-1 cells were assessed by CCK-8 assay, EDU assay and colony formation assay. (e) The apoptosis of Y79 and WERI-Rb-1 cells was analyzed by flow cytometry analysis. (f and g) The migration and invasion of Y79 and WERI-Rb-1 cells were evaluated by wound-healing assay and transwell assay, respectively. (h) The protein levels of PCNA, cleaved-caspase 3, E-cadherin, N-cadherin and vimentin in Y79 and WERI-Rb-1 cells were measured through western blot assay. *** P < 0.001.

Article Snippet: RB cell line WERI-Rb-1 and normal cell line ARPE-19 were provided by Procell (Wuhan, China).

Techniques: Knockdown, Migration, Expressing, Transfection, Quantitative RT-PCR, CCK-8 Assay, EdU Assay, Colony Assay, Flow Cytometry, Wound Healing Assay, Transwell Assay, Western Blot

Journal: Open Medicine

Article Title: circRNF20 aggravates the malignancy of retinoblastoma depending on the regulation of miR-132-3p/PAX6 axis

doi: 10.1515/med-2022-0483

Figure Lengend Snippet: miR-132-3p overexpression inhibited RB cell progression. Y79 and WERI-Rb-1 cells were transfected with miR-NC or miR-132-3p. (a–c) The viability, proliferation and colony formation of Y79 and WERI-Rb-1 cells were evaluated by CCK-8 assay, EDU assay and colony formation assay. (d) The apoptosis of Y79 and WERI-Rb-1 cells was estimated by flow cytometry analysis. (e and f) The migration and invasion of Y79 and WERI-Rb-1 cells were assessed by wound-healing assay and transwell assay. (g) The protein levels of PCNA, cleaved-caspase 3, E-cadherin, N-cadherin and vimentin in Y79 and WERI-Rb-1 cells were measured via western blot assay. *** P < 0.001.

Article Snippet: RB cell line WERI-Rb-1 and normal cell line ARPE-19 were provided by Procell (Wuhan, China).

Techniques: Over Expression, Transfection, CCK-8 Assay, EdU Assay, Colony Assay, Flow Cytometry, Migration, Wound Healing Assay, Transwell Assay, Western Blot

Journal: Open Medicine

Article Title: circRNF20 aggravates the malignancy of retinoblastoma depending on the regulation of miR-132-3p/PAX6 axis

doi: 10.1515/med-2022-0483

Figure Lengend Snippet: circRNF20 regulated RB cell malignant behaviors by miR-132-3p and PAX6. Sh-NC, sh-circRNF20#1, sh-circRNF20#1 + anti-NC, sh-circRNF20#1 + anti-miR-132-3p, sh-circRNF20#1 + vector or sh-circRNF20#1 + PAX6 was transfected into Y79 and WERI-Rb-1 cells. (a) The protein level of PAX6 in Y79 and WERI-Rb-1 cells was examined by qRT-PCR assay. (b–d) The viability, proliferation and colony formation of Y79 and WERI-Rb-1 cells were tested by CCK-8 assay, EDU assay and colony formation assay. (e) The apoptosis of Y79 and WERI-Rb-1 cells was analyzed by flow cytometry analysis. (f and g) The migration and invasion of Y79 and WERI-Rb-1 cells were assessed by wound-healing assay and transwell assay. (h) The protein levels of PCNA, cleaved-caspase 3, E-cadherin, N-cadherin and vimentin in Y79 and WERI-Rb-1 cells were measured by western blot assay. *** P < 0.001.

Article Snippet: RB cell line WERI-Rb-1 and normal cell line ARPE-19 were provided by Procell (Wuhan, China).

Techniques: Plasmid Preparation, Transfection, Quantitative RT-PCR, CCK-8 Assay, EdU Assay, Colony Assay, Flow Cytometry, Migration, Wound Healing Assay, Transwell Assay, Western Blot

Journal: Open Medicine

Article Title: circRNF20 aggravates the malignancy of retinoblastoma depending on the regulation of miR-132-3p/PAX6 axis

doi: 10.1515/med-2022-0483

Figure Lengend Snippet: Effects of circRNF20 derived from RB patients’ serums on RB cell growth, apoptosis and metastasis. (a) The morphology of exosomes was analyzed by TEM. (b) The protein levels of HSP70 and TSG101 were measured by western blot assay. (c) The level of circRNF20 in the exosomes derived from the serums of RB patients and normal controls was detected by qRT-PCR assay. (d) The expression of circRNF20 in Y79 and WERI-Rb-1 cells incubated with RB patients serums’ isolated exosomes was detected by qRT-PCR assay. (e–j) After Y79 and WERI-Rb-1 cells were incubated with the exosomes derived from RB patient serums, cell viability, proliferation, apoptosis, migration and invasion were analyzed. (k) The protein levels of PCNA, cleaved-caspase 3, E-cadherin, N-cadherin and vimentin were measured through western blot assay. *** P < 0.001.

Article Snippet: RB cell line WERI-Rb-1 and normal cell line ARPE-19 were provided by Procell (Wuhan, China).

Techniques: Derivative Assay, Western Blot, Quantitative RT-PCR, Expressing, Incubation, Isolation, Migration

Journal: Annals of Translational Medicine

Article Title: Netrin-1 promotes retinoblastoma-associated angiogenesis

doi: 10.21037/atm-21-5560

Figure Lengend Snippet: Netrin-1 expression is increased in Rb and Rb cell lines. (A) Representative immunohistochemistry images showing netrin-1 expression in Rb samples and normal retina samples; (B) relative netrin-1 mRNA expression in 5 Rb cell lines compared to a normal retina sample; (C) relative netrin-1 protein expression in 5 Rb cell lines compared to a normal retina sample. Rb cells lines: Y79, WERI-Rb-1, NCC-Rbc-57, NCC-Rbc-92, and NCC-Rbc-T1; *, P<0.05; N=5; scale bar is 100 µm.

Article Snippet: Culture and transfection of human Rb cell lines Human Rb cell lines Y79, WERI-Rb-1, NCC-Rbc-57, NCC-Rbc-92, and NCC-Rbc-T1 were purchased from American Type Culture Collection (ATCC, Rockville, MD, USA) and Creative Bioarray (Shirley, NY, USA).

Techniques: Expressing, Immunohistochemistry

Journal: Annals of Translational Medicine

Article Title: Netrin-1 promotes retinoblastoma-associated angiogenesis

doi: 10.21037/atm-21-5560

Figure Lengend Snippet: Silencing netrin-1 by construction of a siRNA plasmid. (A) Illustration of plasmids carrying the siRNA for netrin-1 or a scrambled netrin-1 sequence and a luciferase reporter under a CMV promoter; (B) relative netrin-1 mRNA expression in Rb cells transfected with si-netrin-1 or scrambled plasmid as assessed by RT-qPCR; (C) relative netrin-1 protein expression in Rb cells transfected with si-netrin-1 or scrambled plasmid as assessed by ELISA. Rb cell lines: Y79, WERI-Rb-1, NCC-Rbc-57, NCC-Rbc-92, and NCC-Rbc-T1; *, P<0.05; N=5. IRES, internal ribosome entry site; CMV, cytomegalovirus; RT-qPCR, 1uantitative reverse transcription polymerase chain reaction; ELISA, enzyme-linked immunosorbent assay.

Article Snippet: Culture and transfection of human Rb cell lines Human Rb cell lines Y79, WERI-Rb-1, NCC-Rbc-57, NCC-Rbc-92, and NCC-Rbc-T1 were purchased from American Type Culture Collection (ATCC, Rockville, MD, USA) and Creative Bioarray (Shirley, NY, USA).

Techniques: Plasmid Preparation, Sequencing, Luciferase, Expressing, Transfection, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Reverse Transcription, Polymerase Chain Reaction

Journal: Annals of Translational Medicine

Article Title: Netrin-1 promotes retinoblastoma-associated angiogenesis

doi: 10.21037/atm-21-5560

Figure Lengend Snippet: Netrin-1 depletion compromises angiogenesis without affecting Rb growth in vitro. (A,B) The CCK-8 assay was used to assess cell proliferation in Rb cells transfected with si-netrin-1 or the scrambled plasmid; (C) transfected Rb cells were co-cultured with HUVECs as shown in the illustration; (D,E) the vessel structure formation was analyzed in both Y79 and WERI-Rb-1 cell lines. *, P<0.05; N=5; scale bar represents 100 µm. CCK-8, Cell Counting Kit-8; Rb, retinoblastoma; HUVEC, human umbilical vein endothelial cell; HUVECs, human umbilical vein endothelial cells.

Article Snippet: Culture and transfection of human Rb cell lines Human Rb cell lines Y79, WERI-Rb-1, NCC-Rbc-57, NCC-Rbc-92, and NCC-Rbc-T1 were purchased from American Type Culture Collection (ATCC, Rockville, MD, USA) and Creative Bioarray (Shirley, NY, USA).

Techniques: In Vitro, CCK-8 Assay, Transfection, Plasmid Preparation, Cell Culture, Cell Counting

Journal: Indian Journal of Ophthalmology

Article Title: Retinoblastoma: A review of the molecular basis of tumor development and its clinical correlation in shaping future targeted treatment strategies

doi: 10.4103/IJO.IJO_3172_22

Figure Lengend Snippet: Genes involved in retinoblastoma tumorigenesis identified via multi-omics technologies

Article Snippet: Weri-Rb-1, and NCC-RbC-51 cell lines , Microarray and bioinformatic analyses , qRT-PCR , IL8, IL6, SMAD3, and MYC , Chennai, India Experimental Eye Research, 2020[ ] .

Techniques: Biomarker Discovery, Southern Blot, Methylation Sequencing, Microarray, Western Blot, Immunostaining, Multiplex sample analysis, Clinical Proteomics, Knockdown, Mass Spectrometry, RNA Sequencing, DNA Methylation Assay