Journal: Current Issues in Molecular Biology

Article Title: Genistein–Butein Co-Treatment Suppresses Glycolytic Metabolism and Induces Apoptotic Signaling in PC-3 Prostate Cancer Cells

doi: 10.3390/cimb48030258

Figure Lengend Snippet: Genistein and butein reduce cell viability in prostate cancer cells. ( A ) Chemical structures of GEN and BTN. ( B ) Viability of PC-3 prostate cancer cells and HPrEC following exposure to increasing concentrations of GEN or BTN for 24 or 48 h, evaluated using the MTT assay. ( C ) Cell viability measured by means of trypan blue exclusion under identical treatment conditions. Results are normalized to untreated controls, and IC 50 values were calculated from 48 h treatment data where indicated. Values represent the mean ± SD of at least three independent experiments; * p < 0.05 versus control. HPrEC: normal human prostate epithelial cells; GEN: genistein; BTN: butein; Conc.: concentration.

Article Snippet: Human prostate cancer PC-3 cells and normal human prostate epithelial cells (HPrEC) were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: MTT Assay, Control, Concentration Assay

Journal: Current Issues in Molecular Biology

Article Title: Genistein–Butein Co-Treatment Suppresses Glycolytic Metabolism and Induces Apoptotic Signaling in PC-3 Prostate Cancer Cells

doi: 10.3390/cimb48030258

Figure Lengend Snippet: Combined genistein and butein treatment impairs glycolytic metabolism in PC-3 cells. HPrEC and PC-3 cells were treated with GEN, BTN, or GEN/BTN for 48 h. ( A , B ) Extracellular lactate production and glucose consumption were quantified and normalized to control levels. Enzymatic activities of hexokinase (HK) and pyruvate dehydrogenase (PDH) were assessed following treatment. GEN/BTN co-treatment produced a pronounced suppression of glycolytic activity in PC-3 cells, whereas metabolic parameters in HPrEC cells remained largely unchanged. Data are shown as the mean ± SD (n = 3). Statistical significance is indicated by asterisks (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001). HPrEC: normal human prostate epithelial cells; GEN: genistein; BTN: butein.

Article Snippet: Human prostate cancer PC-3 cells and normal human prostate epithelial cells (HPrEC) were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Control, Produced, Activity Assay

Journal: Current Issues in Molecular Biology

Article Title: Genistein–Butein Co-Treatment Suppresses Glycolytic Metabolism and Induces Apoptotic Signaling in PC-3 Prostate Cancer Cells

doi: 10.3390/cimb48030258

Figure Lengend Snippet: Genistein–butein co-treatment decreases ATP levels and clonogenic survival in PC-3 cells. ( A ) Intracellular ATP content was determined after 48 h exposure to GEN, BTN, or GEN/BTN, and expressed relative to control cells (n = 3 independent experiments). ( B ) Crystal violet staining images illustrating treatment-dependent changes in cell survival. ( C ) Quantitative analysis of crystal violet staining measured at 590 nm. Each dot represents an individual replicate, and data are presented as mean ± SD (total n = 8 pooled from at least three independent experiments). Following GEN/BTN co-treatment in PC-3 cells, a significant reduction in ATP production and cell survival was observed, while normal HPrEC cells showed minimal changes. Results represent the mean ± SD from three independent experiments. Statistical significance is indicated by asterisks (* p < 0.05; ** p < 0.01; **** p < 0.0001). HPrEC: normal human prostate epithelial cells; GEN: genistein; BTN: butein.

Article Snippet: Human prostate cancer PC-3 cells and normal human prostate epithelial cells (HPrEC) were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Control, Staining

Journal: Current Issues in Molecular Biology

Article Title: Genistein–Butein Co-Treatment Suppresses Glycolytic Metabolism and Induces Apoptotic Signaling in PC-3 Prostate Cancer Cells

doi: 10.3390/cimb48030258

Figure Lengend Snippet: Genistein–butein co-treatment promotes apoptosis and suppresses AKT/ERK signaling in PC-3 cells. ( A ) Apoptotic cell populations were evaluated using Annexin V and viability staining after 48 h treatment. Representative dot plots are shown for HPrEC and PC-3 cells. ( B ) Expression levels of hexokinase II and PDH were analyzed by means of Western blotting, with β-actin serving as a loading control. Corresponding densitometric analyses are presented. ( C ) Phosphorylation status of AKT and ERK, along with levels of cleaved caspase-3 and cleaved PARP, was examined by Western blot analysis. GEN/BTN co-treatment selectively inhibited survival signaling and enhanced apoptotic marker cleavage in PC-3 cells. Data are presented as the mean ± SD. Statistical significance is indicated. Statistical significance is indicated by asterisks (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001). HPrEC: normal human prostate epithelial cells; GEN: genistein; BTN: butein; PDH: pyruvate dehydrogenase.

Article Snippet: Human prostate cancer PC-3 cells and normal human prostate epithelial cells (HPrEC) were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Staining, Expressing, Western Blot, Control, Phospho-proteomics, Marker

Journal: International journal of molecular sciences

Article Title: Interactions of CFTR and Arylsulfatase B (ARSB; N-acetylgalactosamine-4-sulfatase) in Prostate Carcinoma.

doi: 10.3390/ijms26094350

Figure Lengend Snippet: Figure 1. CFTR and ARSB in normal and malignant prostate tissue and cells. (A). CFTR expres- sion is reduced in malignant human prostate tissue compared to associated non-malignant tissue (n = 6, p < 0.0001). (B). ARSB activity is reduced in the malignant prostate tissue (n = 6, p < 0.0001). (C). Correlation coefficient r equals 0.98 between the ARSB activity and CFTR expression in the prostate non-malignant and malignant tissues. (D). Both ARSB and CFTR mRNA expression are reduced in three malignant prostate cell lines (LNCaP, TRAMP-C1, and PC3), compared to the normal human prostate epithelial cells (PEC). All p-values are <0.01 (n = 6). (E). Correlation co- efficient between ARSB activity and CFTR expression in the four cell lines in 1D is r equals 0.95. (F). ARSB activity is also reduced in the malignant cell lines, compared to the normal epithelial cells (n = 6, p < 0.00001). Two-tailed, unpaired t-tests, corrected for unequal variance, are used for calculation of p-values. ** represents p ≤0.01 and **** represents p ≤0.0001. [ARSB = arylsulfatase B, N-acetylgalactosamine-4-sulfatase; CFTR = cystic fibrosis transmembrane conductance regulator; PEC = prostate epithelial cells].

Article Snippet: Human prostate epithelial cells (PEC; ATCC®: CRL-2850TM) were obtained and grown under the recommended conditions in Keratinocyte Serum Free Medium (K-SFM) with 0.05 mg/mL bovine pituitary extract (BPE) and 5 ng/mL epidermal growth factor (EGF).

Techniques: Activity Assay, Expressing, Two Tailed Test

Journal: International journal of molecular sciences

Article Title: Interactions of CFTR and Arylsulfatase B (ARSB; N-acetylgalactosamine-4-sulfatase) in Prostate Carcinoma.

doi: 10.3390/ijms26094350

Figure Lengend Snippet: Figure 2. Effects of ARSB and CFTR inhibition. (A). Knockdown of ARSB by siRNA had no effect on CFTR mRNA expression in the normal PEC (n = 6). (B). Inhibition of CFTR by either the chemical inhibitor CFTRinh-172 or CFTR siRNA reduced ARSB expression (p < 0.01, n = 3), but had no impact on expression of GALNS (galactose 6-sulfate sulfatase) and significantly increased expression of Type 1 collagen (Collagen 1; COL1A2) (p = 0.008, p = 0.002). (C). When PC3 cells were treated with CFTR siRNA, ARSB activity declined (p = 0.006, n = 3). The effect of ARSB overexpression was inhibited by CFTR knockdown (p = 0.004, n = 3). (D). ARSB silencing increased total sulfated glycosaminoglycans (GAGs) and chondroitin 4-sulfate (C4S) (p = 0.002, 0.0001, n = 3) in the normal prostate epithelial cells. Similarly, following CFTR silencing, total sulfated (GAG) and C4S increased (p = 0.03, p = 0.0004, n = 3), consistent with inhibition of ARSB when CFTR is silenced. * represents p ≤0.05; ** is for p ≤0.01, *** for p ≤0.001, and **** for p ≤0.0001. [ARSBox = arylsulfatase B overexpression; GAGs = glycosaminoglycans; N.D. = no difference; PEC = prostate epithelial cells; Vcn = vector control].

Article Snippet: Human prostate epithelial cells (PEC; ATCC®: CRL-2850TM) were obtained and grown under the recommended conditions in Keratinocyte Serum Free Medium (K-SFM) with 0.05 mg/mL bovine pituitary extract (BPE) and 5 ng/mL epidermal growth factor (EGF).

Techniques: Inhibition, Knockdown, Expressing, Activity Assay, Over Expression, Plasmid Preparation, Control

Journal: International journal of molecular sciences

Article Title: Interactions of CFTR and Arylsulfatase B (ARSB; N-acetylgalactosamine-4-sulfatase) in Prostate Carcinoma.

doi: 10.3390/ijms26094350

Figure Lengend Snippet: Figure 3. Inhibition of CFTR replicates effects of ARSB knockdown on MMP2, MMP9, and invasive- ness. (A). Expression of MMP2 and MMP9 is significantly increased in malignant prostate tissue, compared to adjacent normal tissue (p = 0.002, p = 0.002; n = 3). (B). Expression of MMP2 and MMP9 is greater in the PC3 cells than in the normal PEC (p = 0.015, p = 0.013; n = 3). (C). Inhibition of CFTR by either CFTRinh-172 or siRNA in the PC3 cells increased the expression of MMP2 (p = 0.005, p = 0.003; n = 3) and MMP9 (p = 0.002, p = 0.002; n = 3). (D,E) ARSB silencing significantly increased MMP2 and MMP9 expression in the PEC cells (p = 0.002, p = 0.05; n = 3) and PC3 cells (p < 0.0001, p = 0.01; n = 3). (F). When ARSB was overexpressed in the PC3 cells, MMP2 and MMP9 expression declined (p = 0.0017, p = 0.026; n = 3). ARSB overexpression reduced the impact of CFTR silencing on MMP9 and MMP2 expression (p = 0.0016, p = 0.0005; n = 3). (G). Invasiveness of the PC3 cells increased when CFTR or ARSB was silenced (p = 0.0014, p = 0.004; n = 3). ARSB overexpression reduced the in- vasiveness to less than the baseline level in the PC3 cells (p = 0.0005; n = 3). The CFTR siRNA-induced increase in invasiveness was reduced when ARSB was overexpressed (p = 0.0014; n = 3). * represents p ≤0.05; ** is for p ≤0.01, *** for p ≤0.001, and **** for p < 0.0001. [ARSBox = ARSB overexpression; MMP = matrix metalloproteinase; PEC = prostate epithelial cells; si = siRNA; Vcn = vector control].

Article Snippet: Human prostate epithelial cells (PEC; ATCC®: CRL-2850TM) were obtained and grown under the recommended conditions in Keratinocyte Serum Free Medium (K-SFM) with 0.05 mg/mL bovine pituitary extract (BPE) and 5 ng/mL epidermal growth factor (EGF).

Techniques: Inhibition, Knockdown, Expressing, Over Expression, Plasmid Preparation, Control

Journal: International journal of molecular sciences

Article Title: Interactions of CFTR and Arylsulfatase B (ARSB; N-acetylgalactosamine-4-sulfatase) in Prostate Carcinoma.

doi: 10.3390/ijms26094350

Figure Lengend Snippet: Figure 4. Effects of ARSB and CFTR on c-Myc and EGFR and BrdU incorporation. (A). EGF (10 ng/mL × 24 h) increased BrdU incorporation in the PC3 cells (p = 0.0004; p = 3). Both ARSB silencing and CFTR silencing further increased the response (p = 0.004, p = 0.0008; n = 3). ARSB overexpression reduced the BrdU incorporation (p = 0.0003; n = 3) and inhibited the effect of CFTR knockdown (p = 0.0009; n = 3). (B). In PC3 cells, mRNA expression of c-Myc and EGFR was greater than in normal PEC (p = 0.004, p = 0.00013; n = 3). (C). In the PEC, silencing ARSB increased mRNA expression of c-Myc (p = 0.004; n = 3) and EGFR (p = 0.00013; n = 3). (D). In PC3 cells, ARSB siRNA increased c-Myc (p = 0.008; n = 3) and EGFR (p = 0.02; n = 3). (E). In PC3 cells, CFTR inhibition by CFTRinh-172 or by siRNA significantly increased c-Myc (p = 0.0002, p = 0.0004; n = 3) and EGFR (p = 0.007, p = 0.0004; n = 3) expression. (F). In PC3 cells, CFTR siRNA significantly increased c-Myc (p = 0.005; n = 3) and EGFR (p = 0.0008; n = 3). ARSB overexpression reduced their expression (p = 0.0025, p = 0.0022; n = 3). In combination with CFTR silencing, ARSB overexpression reduced the CFTR knockdown-induced increases in c-Myc and EGFR (p = 0.066, p = 0.003; n = 3). (G). EGFR protein was significantly increased in the malignant compared to the adjacent non-malignant human prostate tissues (p < 0.0001; n = 6). * represents p ≤0.05; ** is for p ≤0.01, *** for p ≤0.001, and **** for p ≤0.0001. [ARSBox = ARSB overexpression; ARSBsi = ARSB siRNA; CFTRsi = CFTR siRNA; controlsi = control siRNA; EGF = epidermal growth factor; EGFR = epidermal growth factor receptor; PEC = prostate epithelial cells; Vcn = vector control].

Article Snippet: Human prostate epithelial cells (PEC; ATCC®: CRL-2850TM) were obtained and grown under the recommended conditions in Keratinocyte Serum Free Medium (K-SFM) with 0.05 mg/mL bovine pituitary extract (BPE) and 5 ng/mL epidermal growth factor (EGF).

Techniques: BrdU Incorporation Assay, Over Expression, Knockdown, Expressing, Inhibition, Control, Plasmid Preparation