human bc cell lines j82  (iCell Bioscience Inc)

Journal: Molecular medicine reports

Article Title: Propofol modulates the proliferation, invasion and migration of bladder cancer cells through the miR‑145‑5p/TOP2A axis.

doi: 10.3892/mmr.2021.12078

Figure Lengend Snippet: Figure 1. Propofol induces miR‑145‑5p expression in BC cells. (A) Relative expression of miR‑145‑5p in 30 BC samples and paired normal tissues as determined via RT‑qPCR. (B) Relative expression of miR‑145‑5p in BC cell lines (J82 and T24) and SV‑HUC‑1 cells as determined via RT‑qPCR. Relative expression of miR‑145‑5p in (C) J82 and (D) T24 cells at different time points after exposure to 10 µg/ml propofol. (E) J82 and (F) T24 cells were treated with increasing concentrations (0‑20 µg/ml) of propofol for 24, 48 or 72 h, then cell viability was evaluated. Each assay was performed in triplicate and data are presented as the mean ± SD. *P<0.05. BC, breast cancer; miR, microRNA; RT‑qPCR, reverse transcription‑quantitative PCR.

Article Snippet: The human BC cell lines J82 and T24, as well as the SV40 immortalized human uroepithelial cell line SV‐HUC‐1, were purchased from American Type Culture Collection, and cultured in RPMI‐1640 medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBC (HyClone; Cytiva) at 37 ̊C in a humidified incubator containing 5% CO2.

Techniques: Expressing

Journal: Molecular medicine reports

Article Title: Propofol modulates the proliferation, invasion and migration of bladder cancer cells through the miR‑145‑5p/TOP2A axis.

doi: 10.3892/mmr.2021.12078

Figure Lengend Snippet: Figure 2. Propofol inhibits the proliferation and migration of bladder cancer cells by regulating miR‑145‑5p expression. (A) J82 and T24 cells were transduced with anti‑miR‑145‑5p, anti‑miR‑NC, EV or miR‑145‑5p, then the relative expression of miR‑145‑5p was evaluated via RT‑qPCR. (B) J82 and (C) and T24 cells were transduced with anti‑miR‑145‑5p, anti‑miR‑NC or miR‑145‑5p and exposed to 10 µg/ml propofol, then relative miR‑145‑5p expression was evaluated via RT‑qPCR. (D) J82 and (E) T24 cells were transduced with anti‑miR‑145‑5p, anti‑miR‑NC or miR‑145‑5p and exposed to 10 µg/ml propofol, then cell viability was evaluated at 24, 48 and 72 h post‑drug treatment. (F) J82 and (G) T24 cells were transduced with anti‑miR‑145‑5p, anti‑miR‑NC or miR‑145‑5p and exposed to 10 µg/ml propofol, then cells were used for wound‑healing assays. Scale bar, 500 µm. Each assay was performed in triplicate and data are presented as the mean ± SD. *P<0.05. miR, microRNA; NC, negative control; EV, empty vector; RT‑qPCR, reverse transcription‑quantitative PCR.

Article Snippet: The human BC cell lines J82 and T24, as well as the SV40 immortalized human uroepithelial cell line SV‐HUC‐1, were purchased from American Type Culture Collection, and cultured in RPMI‐1640 medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBC (HyClone; Cytiva) at 37 ̊C in a humidified incubator containing 5% CO2.

Techniques: Migration, Expressing, Transduction, Negative Control, Plasmid Preparation

Journal: Molecular medicine reports

Article Title: Propofol modulates the proliferation, invasion and migration of bladder cancer cells through the miR‑145‑5p/TOP2A axis.

doi: 10.3892/mmr.2021.12078

Figure Lengend Snippet: Figure 3. Identification of TOP2A as a target of miR‑145‑5p. (A) Top 10 mRNA transcripts upregulated and downregulated in GSE76211 are shown in a heat map. (B) Potential binding site for TOP2A and miR‑145‑5p as identified by TargetScan. (C) J82 and T24 cells were transduced with miR‑145‑5p or EV lenti virus control, then relative TOP2A expression was evaluated via reverse transcription‑quantitative PCR. (D) J82 and (E) T24 cells were transfected with EV or miR‑145‑5p and TOP2A 3'UTR‑WT or TOP2A 3'UTR‑MUT, then cells were used for luciferase reporter assays. Each assay was performed in triplicate and data are presented as the mean ± SD. *P<0.05. N, normal; T, tumor; miR, microRNA; EV, empty vector; TOP2A, topoisomerase II α; 3'UTR, 3‑untranslated region; WT, wild‑type; MUT, mutant.

Article Snippet: The human BC cell lines J82 and T24, as well as the SV40 immortalized human uroepithelial cell line SV‐HUC‐1, were purchased from American Type Culture Collection, and cultured in RPMI‐1640 medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBC (HyClone; Cytiva) at 37 ̊C in a humidified incubator containing 5% CO2.

Techniques: Binding Assay, Transduction, Virus, Control, Expressing, Transfection, Luciferase, Plasmid Preparation, Mutagenesis

Journal: Molecular medicine reports

Article Title: Propofol modulates the proliferation, invasion and migration of bladder cancer cells through the miR‑145‑5p/TOP2A axis.

doi: 10.3892/mmr.2021.12078

Figure Lengend Snippet: Figure 4. Propofol suppresses TOP2A expression in BC cells. (A) Relative TOP2A expression in 30 BC samples and paired normal tissues as determined via RT‑qPCR. (B) Relative TOP2A expression between BC samples and paired normal tissues as determined via western blot analysis. (C) Relative TOP2A expression in different cell lines as determined via RT‑qPCR. (D) Relative TOP2A expression in different cell lines as determined via western blot analysis. J82 and T24 cells were treated with 10 µg/ml propofol or an equal volume of DMSO as control, then relative TOP2A expression was evaluated via (E) RT‑qPCR and (F) western blot analysis. (G) Spearman correlation analysis of miR‑145‑5p and TOP2A expression in patients with BC. Each assay was performed in triplicate and data are presented as the mean ± SD. *P<0.05. BC, bladder cancer; N, normal; T, tumor; miR, microRNA; TOP2A, topoisomerase II α; RT‑qPCR, reverse transcription‑quantitative PCR.

Article Snippet: The human BC cell lines J82 and T24, as well as the SV40 immortalized human uroepithelial cell line SV‐HUC‐1, were purchased from American Type Culture Collection, and cultured in RPMI‐1640 medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBC (HyClone; Cytiva) at 37 ̊C in a humidified incubator containing 5% CO2.

Techniques: Expressing, Western Blot, Control

Journal: Molecular medicine reports

Article Title: Propofol modulates the proliferation, invasion and migration of bladder cancer cells through the miR‑145‑5p/TOP2A axis.

doi: 10.3892/mmr.2021.12078

Figure Lengend Snippet: Figure 5. Propofol exerts antitumor effects via the miR‑145‑5p/TOP2A axis. (A) J82 and T24 cells were transduced with anti‑miR‑NC or anti‑miR‑145‑5p, then relative TOP2A expression was evaluated via RT‑qPCR. (B) J82 and T24 cells were transfected with si‑NC or si‑TOP2A, then relative TOP2A expression was evaluated via RT‑qPCR. (C) J82 and (D) T24 cells were transduced with anti‑miR‑NC, anti‑miR‑145‑5p, si‑NC or si‑TOP2A, then treated with 10 µg/ml propofol or an equal volume of DMSO as control. Relative TOP2A expression was evaluated via RT‑qPCR. (E) J82 and (F) T24 cells were transduced with anti‑miR‑NC, anti‑miR‑145‑5p, si‑NC or si‑TOP2A, then treated with 10 µg/ml propofol or an equal volume of DMSO as control. Relative TOP2A expression was evaluated via western blotting. Each assay was performed in triplicate and data are presented as the mean ± SD. *P<0.05. miR, microRNA; si, small interfering RNA; NC, negative control; TOP2A, topoisomerase II α; RT‑qPCR, reverse transcription‑quantitative PCR.

Article Snippet: The human BC cell lines J82 and T24, as well as the SV40 immortalized human uroepithelial cell line SV‐HUC‐1, were purchased from American Type Culture Collection, and cultured in RPMI‐1640 medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBC (HyClone; Cytiva) at 37 ̊C in a humidified incubator containing 5% CO2.

Techniques: Transduction, Expressing, Transfection, Control, Western Blot, Small Interfering RNA, Negative Control