c4-2b  (ATCC)

Journal: Cancers

Article Title: Growth Hormone-Releasing Hormone (GHRH) Antagonist Peptides Combined with PI3K Isoform Inhibitors Enhance Cell Death in Prostate Cancer

doi: 10.3390/cancers17101643

Figure Lengend Snippet: MIA-602 and -690 GHRH antagonists increase cell death in PCa/CRPC/NEPC cells. Trypan blue exclusion assay showed significantly higher cell death (72 h) in MIA-602 (02) and -690 (90) (5 μM)-treated LNCaP (PCa), 22Rv1 (CRPC), and H660 LASCPC (NEPC) cells compared to control (—)-treated cells. There was no increased cell death in PC3 or DU145 (CRPC). P values are shown above the bars (bold numbers indicate significance).

Article Snippet: Human AR+ androgen-sensitive PCa (LNCaP), AR+ CRPC (22Rv1), AR—CRPC (PC3, DU145), AR—NEPC (NCI-H660, LASCPC), and human non-cancer RWPE-1 (prostate epithelial) cells were obtained from the American Type Culture Collection (ATCC) and were used within 6 months of the resuscitation of the original cultures.

Techniques: Trypan Blue Exclusion Assay, Control

Journal: Cancers

Article Title: Growth Hormone-Releasing Hormone (GHRH) Antagonist Peptides Combined with PI3K Isoform Inhibitors Enhance Cell Death in Prostate Cancer

doi: 10.3390/cancers17101643

Figure Lengend Snippet: Searching for a drug combination with MIA-602 and -690 GHRH antagonist peptides that will increase cell death in PCa/CRPC/NEPC. ( A ) Trypan blue exclusion assay shows that anti-mitotic docetaxel (D; 0.25 nM LNCaP, 22Rv1; 1 nM PC3) + MIA-602 (02) or -690 (90) (5 μM) increases cell death in LNCaP, 22Rv1, and PC3 cells compared to D, 02/90, and control cells. Positive control is D + Bcl-2 (B) inhibitor ABT-737 (1 μM). ( B ) Bcl-2 (B) inhibitor ABT-737 (1 μM) + MIA-602 or -690 partially increases cell death in LNCaP and PC3 but not in 22Rv1 cells. ( C ) Pan-PI3K inhibitor LY294002 (LY, 10 μM) + MIA-602 or -690 significantly increases cell death in LNCaP, 22Rv1, and PC3 compared to LY, 02/90, and control cells. P values are shown near the bars (bold numbers indicate significance).

Article Snippet: Human AR+ androgen-sensitive PCa (LNCaP), AR+ CRPC (22Rv1), AR—CRPC (PC3, DU145), AR—NEPC (NCI-H660, LASCPC), and human non-cancer RWPE-1 (prostate epithelial) cells were obtained from the American Type Culture Collection (ATCC) and were used within 6 months of the resuscitation of the original cultures.

Techniques: Trypan Blue Exclusion Assay, Control, Positive Control

Journal: Cancers

Article Title: Growth Hormone-Releasing Hormone (GHRH) Antagonist Peptides Combined with PI3K Isoform Inhibitors Enhance Cell Death in Prostate Cancer

doi: 10.3390/cancers17101643

Figure Lengend Snippet: PI3Kα or β isoform inhibitors + MIA-602 or -690 increase cell death in PCa/CRPC/NEPC cells. ( A ) Trypan blue exclusion assay shows PI3Kαi (αi, 10 μM) + MIA-602 (02) or -690 (90) (5 μM) significantly increase cell death in 22Rv1 compared to αi, 02/90, and control cells. In LNCaP, PI3Kαi does not increase MIA-602 or -690 cell death. PI3Kβi (βi, 2.5 μM) + MIA-602 or -690 significantly increase cell death in LNCaP compared to βi, 02/90, and control cells, but only slightly increase cell death in 22Rv1 (βi, 10 μM). ( B ) PI3Kαi + MIA-602 or -690 significantly increase cell death in PC3 but partially in DU145 (90 + αi). PI3Kβi (10 μM) significantly increases MIA-602 or -690 cell death in DU145 but not in PC3. ( C ) PI3Kαi + MIA-602 or -690 significantly increase cell death in H660 and LASCPC whereas PI3Kβi (10 μM) + MIA-602 or -690 slightly increase cell death in LASCPC but not in H660. P values are shown near the bars (bold numbers indicate significance).

Article Snippet: Human AR+ androgen-sensitive PCa (LNCaP), AR+ CRPC (22Rv1), AR—CRPC (PC3, DU145), AR—NEPC (NCI-H660, LASCPC), and human non-cancer RWPE-1 (prostate epithelial) cells were obtained from the American Type Culture Collection (ATCC) and were used within 6 months of the resuscitation of the original cultures.

Techniques: Trypan Blue Exclusion Assay, Control

Journal: Cancers

Article Title: Growth Hormone-Releasing Hormone (GHRH) Antagonist Peptides Combined with PI3K Isoform Inhibitors Enhance Cell Death in Prostate Cancer

doi: 10.3390/cancers17101643

Figure Lengend Snippet: MIA-602/690 alone and + PI3K isoform inhibitors alter multiple signaling pathways and AR expression. ( A ) Western blot analysis showed that MIA-602 (02, 5 μM) + PI3Kαi (αi, 10 μM) decreased PI3Kα, PI3Kβ, and AKT in 22Rv1 and PC3 (P-AKT increased in PC3 24 h); MIA-602 alone decreased PI3Kα in 22Rv1. MIA-602 decreased P-ERK in 22Rv1 (72 h) (PI3Kαi increased P-ERK). MIA-602 alone and MIA-602 + PI3Kαi decreased the total ERK in 22Rv1 (72 h). In PC3, there was a switch from P-ERK1 to P-ERK2 with MIA-602 and MIA-602 + PI3Kαi (24, 72 h). No clear differences were noted in GHRHR and cl-PARP. ( B ) In LNCaP, MIA-602/690 alone and MIA-602/690 + PI3Kβi (βi, 2.5 μM) (24 h) decreased Mcl-1L (anti-apoptosis) and increased Mcl-1S (pro-apoptosis). No clear difference in the apoptosis marker cl-PARP was noted. PI3Kβi alone and MIA-602/690 + PI3Kβi decreased proliferation markers E2F1 and cyclin A. MIA-690 + PI3Kβi decreased PI3Kα, PI3Kβ, and P/T-AKT. AR was strongly decreased with MIA-602/690 alone. ( C ) In LNCaP, treatment with MIA-602/690 over time (4–72 h) decreased Mcl-1L, PI3Kα, PI3Kβ, P-AKT, and AR. No clear differences were noted in GHRHR. Protein refers to the Coomassie blue stain of blots after all immunological analysis was completed. Quantification values (divided by protein [p]) for PI3Kα, PI3Kβ, AKT, ERK, Mcl-1L, Mcl-1S, E2F1, cyclin A, and AR are shown below specific bands with control = 1. The ratio of P/T (total) AKT and ERK values are also shown.

Article Snippet: Human AR+ androgen-sensitive PCa (LNCaP), AR+ CRPC (22Rv1), AR—CRPC (PC3, DU145), AR—NEPC (NCI-H660, LASCPC), and human non-cancer RWPE-1 (prostate epithelial) cells were obtained from the American Type Culture Collection (ATCC) and were used within 6 months of the resuscitation of the original cultures.

Techniques: Protein-Protein interactions, Expressing, Western Blot, Marker, Staining, Control

Journal: Cancers

Article Title: Growth Hormone-Releasing Hormone (GHRH) Antagonist Peptides Combined with PI3K Isoform Inhibitors Enhance Cell Death in Prostate Cancer

doi: 10.3390/cancers17101643

Figure Lengend Snippet: MIA-602Ac and -690Ac are clinically relevant forms with similar effects on signaling pathways. ( A ) Trypan blue exclusion assay showed that MIA-602Ac (02) or -690Ac (90) (5 μM) + PI3Kβi (βi, 25 nM) (LNCaP) or PI3Kαi (αi, 10 μM) (22Rv1) significantly increased cell death compared to βi/αi, 02/90, and control cells. p values are shown near the bars. ( B ) Western blot analysis in LNCaP showed that MIA-602Ac or -690Ac (5 μM) + PI3Kβi (25 nM) increased cl-PARP better than either one alone (24, 48 h). MIA-602Ac/690Ac decreased Mcl-1L (apoptosis), E2F1, cyclin A (proliferation), GHRHR (stronger with 90 at 24 h), and AR, and increased Mcl-1S. ERK was increased by PI3Kβi. ( C ) In 22Rv1, Western blot results showed that MIA-602Ac/690Ac + PI3Kαi (10 μM) had similar changes compared to LNCaP, with MIA-690Ac demonstrating stronger effects. Protein refers to the Coomassie blue staining of blots after all immunological analysis was completed.

Article Snippet: Human AR+ androgen-sensitive PCa (LNCaP), AR+ CRPC (22Rv1), AR—CRPC (PC3, DU145), AR—NEPC (NCI-H660, LASCPC), and human non-cancer RWPE-1 (prostate epithelial) cells were obtained from the American Type Culture Collection (ATCC) and were used within 6 months of the resuscitation of the original cultures.

Techniques: Protein-Protein interactions, Trypan Blue Exclusion Assay, Control, Western Blot, Staining

Journal: Brazilian Journal of Medical and Biological Research

Article Title: Overexpression of SERPINA3 inhibits castration-resistant prostate cancer progression by enhancing M1 macrophage recruitment via CXCL2 upregulation

doi: 10.1590/1414-431X2025e14445

Figure Lengend Snippet: Negative association between SERPINA3 expression and castration-resistant prostate cancer (CRPC). A , Differentially expressed genes identified by Venn analysis in the three CRPC-related datasets ( GSE101607 , GSE32269 , and GSE109708 ) and the expression of SERPINA3 in GSE32269 and GSE109708 datasets. B , Profile of SERPINA3 expression in the Michigan 2012 PCa dataset. C , Profile of SERPINA3 expression in Cambridge 2012 PCa dataset. D , Expression of SERPINA3 in five PCa cell lines (LNCAP, VCAP, C4-2B, PC3, and DU145) detected by western blotting. E , Protein expression of SERPINA3 was detected in LNCAP cells treated with androgen-deprived medium for different durations. F , Representative images of SERPINA3 staining in tissue samples from patients with hormone therapy-sensitive PCa as well as patients with CRPC (n=3:3). Data are reported as means±SE and were analyzed by two-tailed unpaired Student's t -test. *P<0.05, **P<0.01, ***P<0.001. Scale bar, 50 μm. AR: androgen receptor.

Article Snippet: The human PCa cell lines LNCAP, VCAP, C4-2B, PC3, and DU145 and the human monocytic leukemia cell line THP-1 were purchased from American Type Culture Collection.

Techniques: Expressing, Western Blot, Staining, Two Tailed Test

Journal: Pharmaceuticals

Article Title: Is Copper-61 the New Gallium-68? Automation and Preclinical Proof-of-Concept of 61 Cu-Based Radiopharmaceuticals for Prostate Cancer Imaging

doi: 10.3390/ph18040469

Figure Lengend Snippet: Cellular uptake of ( A ) [ 61 Cu]Cu-DOTAGA-PSMA-I&T and ( B ) [ 61 Cu]Cu-NODAGA-PSMA-I&T in PSMA-positive (LNCaP) and PSMA-negative (DU145) cells. Data are presented as mean ± SEM (N = 2–5).

Article Snippet: The human PCa cell lines LNCaP (PSMA-positive) (American Type Culture Collection (ATCC), CRL-1740; Manassas, VA, USA) and DU145 (PSMA-negative) (kindly provided by Dr. Carmen Jerónimo Lab at IPO-Porto) were maintained at 37 °C in a humidified incubator enriched with 5% carbon dioxide.

Techniques:

Journal: Pharmaceuticals

Article Title: Is Copper-61 the New Gallium-68? Automation and Preclinical Proof-of-Concept of 61 Cu-Based Radiopharmaceuticals for Prostate Cancer Imaging

doi: 10.3390/ph18040469

Figure Lengend Snippet: Representative axial (upper row) and coronal (lower row) whole-body PET/MR images acquired at 1 h (left column) and 4 h (right column) post-injection of [ 61 Cu]Cu-NODAGA-PSMA-I&T into the tail vein of LNCaP tumor-bearing mice. Images were normalized to % ID/g.

Article Snippet: The human PCa cell lines LNCaP (PSMA-positive) (American Type Culture Collection (ATCC), CRL-1740; Manassas, VA, USA) and DU145 (PSMA-negative) (kindly provided by Dr. Carmen Jerónimo Lab at IPO-Porto) were maintained at 37 °C in a humidified incubator enriched with 5% carbon dioxide.

Techniques: Positron Emission Tomography-Magnetic Resonance Imaging, Injection