human normal retinal pigment epithelial cells  (ATCC)

Journal: Clinics

Article Title: microRNA-181a-5p impedes the proliferation, migration, and invasion of retinoblastoma cells by targeting the NRAS proto-oncogene

doi: 10.1016/j.clinsp.2022.100026

Figure Lengend Snippet: MiR-181a-5p is downregulated in RB tissues and cell lines. (A) Quantitative reverse Transcription-Polymerase Chain Reaction (qRT-PCR) showed that miR-181a-5p was significantly downregulated in RB tissues compared with the normal retinal tissues. (B) qRT-PCR showed that miR-181a-5p was significantly downregulated in RB cell lines compared to the normal retinal pigment epithelial cells. The expression of miR-181a-5p was normalized to that of the U6 small nuclear RNA (snRNA). The data are shown as the mean ± standard deviation from three independent experiments. **p < 0.01 and ***p < 0.001.

Article Snippet: Human normal retinal pigment epithelial cells (ARPE-19) and RB cell lines (HXO-RB44, SO-Rb50, Y79, and WERI-RB-1) were procured from the American Type Culture Collection (ATCC, Rockville, MD, USA) or China Center for Type Culture Collection (CCTCC, Wuhan, China).

Techniques: Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Expressing, Standard Deviation

Journal: Experimental and Therapeutic Medicine

Article Title: Knockdown of lncRNA NEAT1 expression inhibits cell migration, invasion and EMT by regulating the miR-24-3p/LRG1 axis in retinoblastoma cells

doi: 10.3892/etm.2021.9798

Figure Lengend Snippet: NEAT1 knockdown significantly suppresses cell migration, invasion and EMT in RB cells. sh-NEAT1 or sh-NC were transfected into Y79 and WERI-RB1 cells. Expression of NEAT1 in (A) Y79 and (B) WERI-RB1 cells was examined via reverse transcription-quantitative PCR. (C) Migration and (D) invasion of Y79 and WERI-RB1 cells were evaluated via a Transwell assay. Protein expression levels of MMP9, N-cadherin and E-cadherin in (E) Y79 and (F) WERI-RB1 cells were detected via western blotting. ** P<0.01 and *** P<0.001 vs. sh-NC. miR, microRNA; NEAT1, nuclear paraspeckle assembly transcript 1; RB, retinoblastoma; MMP, matrix metalloproteinase; sh, short hairpin RNA; NC, negative control; EMT, epithelial-to-mesenchymal transition.

Article Snippet: Human RB cell lines (Y79 and WERI-RB1) and normal human retinal pigment epithelial cell line (ARPE-19) were obtained from the American Type Culture Collection.

Techniques: Knockdown, Migration, Transfection, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Transwell Assay, Western Blot, shRNA, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: Knockdown of lncRNA NEAT1 expression inhibits cell migration, invasion and EMT by regulating the miR-24-3p/LRG1 axis in retinoblastoma cells

doi: 10.3892/etm.2021.9798

Figure Lengend Snippet: NEAT1 directly targets miR-24-3p and negatively regulates miR-24-3p expression in RB cells. (A) Predicted binding sites between miR-24-3p and NEAT1. Luciferase activities in (B) Y79 and (C) WERI-RB1 cells co-transfected with miR-24-3p or miR-NC and WT-NEAT1 or MUT-NEAT1. ** P<0.01 vs. miR-NC, *** P<0.001 vs. miR-NC. The expression level of miR-24-3p in (D) Y79 and (E) WERI-RB1 cells transfected with pcDNA, pcDNA-NEAT1, sh-NC or sh-NEAT1 was detected via reverse transcription-quantitative PCR. ** P<0.01 vs. pcDNA, ## P<0.01 vs. sh-NC. miR, microRNA; NEAT1, nuclear paraspeckle assembly transcript 1; RB, retinoblastoma; sh, short hairpin RNA; NC, negative control; EMT, epithelial-to-mesenchymal transition; WT, wild-type; MUT, mutant.

Article Snippet: Human RB cell lines (Y79 and WERI-RB1) and normal human retinal pigment epithelial cell line (ARPE-19) were obtained from the American Type Culture Collection.

Techniques: Expressing, Binding Assay, Luciferase, Transfection, Reverse Transcription, Real-time Polymerase Chain Reaction, shRNA, Negative Control, Mutagenesis

Journal: Experimental and Therapeutic Medicine

Article Title: Knockdown of lncRNA NEAT1 expression inhibits cell migration, invasion and EMT by regulating the miR-24-3p/LRG1 axis in retinoblastoma cells

doi: 10.3892/etm.2021.9798

Figure Lengend Snippet: NEAT1 represses RB cell migration, invasion and EMT via targeting miR-24-3p. Y79 and WERI-RB1 cells were treated with miR-24-3p, miR-NC, sh-NEAT1, sh-NC, sh-NEAT1 + anti NC or sh-NEAT1 + anti miR-24-3p. (A) Migrated and (B) invaded cell numbers of Y79 and WERI-RB1 cells were evaluated by Transwell assay. Protein expression levels of MMP9, N-cadherin and E-cadherin in (C) Y79 and (D) WERI-RB1 cells were detected by western blot analysis. * P<0.05, ** P<0.01 and *** P<0.001. miR, microRNA; NEAT1, nuclear paraspeckle assembly transcript 1; RB, retinoblastoma; MMP, matrix metalloproteinase; sh, short hairpin RNA; NC, negative control; EMT, epithelial-to-mesenchymal transition.

Article Snippet: Human RB cell lines (Y79 and WERI-RB1) and normal human retinal pigment epithelial cell line (ARPE-19) were obtained from the American Type Culture Collection.

Techniques: Migration, Transwell Assay, Expressing, Western Blot, shRNA, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: Knockdown of lncRNA NEAT1 expression inhibits cell migration, invasion and EMT by regulating the miR-24-3p/LRG1 axis in retinoblastoma cells

doi: 10.3892/etm.2021.9798

Figure Lengend Snippet: NEAT1 affects RB cell migration, invasion and EMT by upregulating LRG1 expression via the sponging of miR-24-3p. Y79 and WERI-RB1 cells were divided into six groups and transfected with the following: pcDNA-LRG1, pcDNA, pcDNA-LRG1 + miR-24-3p, pcDNA-LRG1 + miR-NC, pcDNA-LRG1 + miR-24-3p + pcDNA-NEAT1 or pcDNA-LRG1 + miR-24-3p + pcDNA. (A) Migration and (B) invasion of Y79 and WERI-RB1 cells were evaluated via a Transwell assay. MMP9, N-cadherin and E-cadherin protein levels in (C) Y79 and (D) WERI-RB1 cells were determined via a western blot assay. * P<0.05, ** P<0.01 and *** P<0.001. miR, microRNA; NEAT1, nuclear paraspeckle assembly transcript 1; RB, retinoblastoma; MMP, matrix metalloproteinase; sh, short hairpin RNA; NC, negative control; EMT, epithelial-to-mesenchymal transition; LRG1, leucine-rich α-2-glycoprotein.

Article Snippet: Human RB cell lines (Y79 and WERI-RB1) and normal human retinal pigment epithelial cell line (ARPE-19) were obtained from the American Type Culture Collection.

Techniques: Migration, Expressing, Transfection, Transwell Assay, Western Blot, shRNA, Negative Control