human prostate cancer cell line c4 2b  (ATCC)

Journal: Antioxidants

Article Title: Quercetin Inhibits AKT Ser473 Phosphorylation and Disrupts AKT–Androgen Receptor Signaling in Castration-Resistant Prostate Cancer Cells

doi: 10.3390/antiox15030393

Figure Lengend Snippet: The effect of QRC on the proliferation and cell viability of CRPC cell lines. C4-2B and 22Rv1 cells were cultured in the presence of increasing concentrations of QRC (0, 6.25, 12.5, 25 and 50 μM), and images were taken each hour up to 48 h using the IncuCyte S3 system. The effect of QRC on cell proliferation is shown for C4-2B ( A ) and 22Rv1 ( B ) cells during the 48 h analysis, with the effect at 48 h of treatment presented as a bar plot. The effect of QRC on cell death (based on the incorporation of a live-cell-impermeable nucleic acid stain) is shown for C4-2B ( C ) and 22Rv1 ( D ) cells during the 48 h analysis, with the effect at 48 h of treatment presented as a bar plot. The data represent the means ± SEM of four independent experiments analyzed by a one-way analysis of variance and Dunn’s multiple comparison post-test (**** p ≤ 0.0001, *** p ≤ 0.001, ** p ≤ 0.01, * p ≤ 0.05).

Article Snippet: Human prostate cancer cells C4-2B (#CRL-3315, ATCC, Manassas, VA, USA) and 22Rv1 (#05092802, ECACC, Salisbury, UK) were routinely grown in RPMI 1640 medium, supplemented with 2 mM Glutagro (25-015-CI, Corning, Corning, NY, USA), 10% fetal bovine serum (HC.SV30160.03, Cytiva, Marlborough, MA, USA), and 1% penicillin–streptomycin (HC.SV30010, Cytiva), and maintained at 37 °C with 5% CO 2 in a humidified incubator.

Techniques: Cell Culture, Staining, Comparison

Journal: Antioxidants

Article Title: Quercetin Inhibits AKT Ser473 Phosphorylation and Disrupts AKT–Androgen Receptor Signaling in Castration-Resistant Prostate Cancer Cells

doi: 10.3390/antiox15030393

Figure Lengend Snippet: The effect of QRC on AKT activation and AR expression in CRPC cell lines. A Western blot analysis of the levels of AKT phosphorylated at S473 (P-AKT S473), total AKT (T-AKT), AR phosphorylated at S210/213 and total AR (AR) in total protein extracts from C4-2B ( A ) and 22Rv1 ( B ) cells incubated with or without 25 μM QRC for 1 h and 24 h. GAPDH was analyzed as the loading control. The Western blot images were quantified and data presented in plots as follows: pAKT/AKT, pAR/AR, and AR/GAPDH. The data represent the means ± SEM of at least three independent experiments analyzed by Student’s t -test (** p ≤ 0.01, * p ≤ 0.05). ns: not statistically significant.

Article Snippet: Human prostate cancer cells C4-2B (#CRL-3315, ATCC, Manassas, VA, USA) and 22Rv1 (#05092802, ECACC, Salisbury, UK) were routinely grown in RPMI 1640 medium, supplemented with 2 mM Glutagro (25-015-CI, Corning, Corning, NY, USA), 10% fetal bovine serum (HC.SV30160.03, Cytiva, Marlborough, MA, USA), and 1% penicillin–streptomycin (HC.SV30010, Cytiva), and maintained at 37 °C with 5% CO 2 in a humidified incubator.

Techniques: Activation Assay, Expressing, Western Blot, Incubation, Control

Journal: Antioxidants

Article Title: Quercetin Inhibits AKT Ser473 Phosphorylation and Disrupts AKT–Androgen Receptor Signaling in Castration-Resistant Prostate Cancer Cells

doi: 10.3390/antiox15030393

Figure Lengend Snippet: The effect of QRC on AR signaling in CRPC cell lines. ( A ) C4-2B and 22Rv1 cells were incubated in the presence or absence of 25 μM QRC for 24 h, and the culture medium was used for the analysis of the levels of PSA by dot blot. PSA was quantified after normalization against the concentration of total proteins in the medium, and data were plotted. ( B ) The real-time RT-PCR analysis of AR mRNA expression in C4-2B and 22Rv1 cells in response to 25 μM QRC for 24 h. ( C ) Representative images of the confocal analysis of the effect of QRC on AR nuclear distribution using DAPI as the nuclear marker. ( D ) For quantification, the intensity of the AR nuclear signal was normalized against DAPI. The data represent the means ± SEM of at least three independent experiments analyzed by Student’s t -test (**** p ≤ 0.0001, ** p ≤ 0.01). ns: not statistically significant.

Article Snippet: Human prostate cancer cells C4-2B (#CRL-3315, ATCC, Manassas, VA, USA) and 22Rv1 (#05092802, ECACC, Salisbury, UK) were routinely grown in RPMI 1640 medium, supplemented with 2 mM Glutagro (25-015-CI, Corning, Corning, NY, USA), 10% fetal bovine serum (HC.SV30160.03, Cytiva, Marlborough, MA, USA), and 1% penicillin–streptomycin (HC.SV30010, Cytiva), and maintained at 37 °C with 5% CO 2 in a humidified incubator.

Techniques: Incubation, Dot Blot, Concentration Assay, Quantitative RT-PCR, Expressing, Marker

Journal: Antioxidants

Article Title: Quercetin Inhibits AKT Ser473 Phosphorylation and Disrupts AKT–Androgen Receptor Signaling in Castration-Resistant Prostate Cancer Cells

doi: 10.3390/antiox15030393

Figure Lengend Snippet: Enzalutamide (Enz) does not enhance the anti-proliferative effect of QRC on CRPC cell lines. C4-2B cells were cultured with 20 μM Enz and 22Rv1 cells with 40 μM Enz, and both cell lines were treated with increasing concentrations of QRC (0, 6.25, 12.5, 25 and 50 μM) up to 48 h to establish any additive effect. The plots show the effect of QRC alone (green bars) and the effect of Enz plus QRC (blue bars) after 48 h of treatment. The data represent the means ± SEM of at least three independent experiments analyzed by Student’s t -test (* p ≤ 0.05; ns: not statistically significant).

Article Snippet: Human prostate cancer cells C4-2B (#CRL-3315, ATCC, Manassas, VA, USA) and 22Rv1 (#05092802, ECACC, Salisbury, UK) were routinely grown in RPMI 1640 medium, supplemented with 2 mM Glutagro (25-015-CI, Corning, Corning, NY, USA), 10% fetal bovine serum (HC.SV30160.03, Cytiva, Marlborough, MA, USA), and 1% penicillin–streptomycin (HC.SV30010, Cytiva), and maintained at 37 °C with 5% CO 2 in a humidified incubator.

Techniques: Cell Culture