Journal: Oncotarget

Article Title: The tumor suppressive role of CAMK2N1 in castration-resistant prostate cancer

doi:

Figure Lengend Snippet: (A) Representative examples of immunohistochemical staining for CAMK2N1 in each of the clinical stages of prostate cancers as indicated. (B) Quantification of CAMK2N1 relative immunostaining intensity for each clinical stage of prostate cancers. Data is shown as mean ± SEM for N as indicated in the figure in parethesis as shown. (C) CAMK2N1 mRNA determined by quantitative PCR. Comparison was made between normal and tumorous prostate samples. (D) QT-PCR analysis of CAMK2N1 expression in prostate cell lines (RWPE1, LNCaP, DU145, PC3).

Article Snippet: 2 × 10 6 DU145 cells and PC3 cells expressing CAMK2N1 were implanted subcutaneously into 4-6-week-old athymic female nude mice purchased from Beijing HFK Bio-Technology.co., LTD. Tumor growth was measured using a digital caliper every 5 days for 4-5 weeks.

Techniques: Immunohistochemical staining, Staining, Immunostaining, Real-time Polymerase Chain Reaction, Comparison, Expressing

Journal: Oncotarget

Article Title: The tumor suppressive role of CAMK2N1 in castration-resistant prostate cancer

doi:

Figure Lengend Snippet: DU145 and PC3 cells stably overexpressed or knocked down CAMK2N1. Cells were analyzed for cell proliferation by (A-C) MTT, cell cycle by (D-E) FACS and oncogenic growth by (F-K) Colony-forming assay. Data is shown as mean ± SEM for N > 5 separate experiments.

Article Snippet: 2 × 10 6 DU145 cells and PC3 cells expressing CAMK2N1 were implanted subcutaneously into 4-6-week-old athymic female nude mice purchased from Beijing HFK Bio-Technology.co., LTD. Tumor growth was measured using a digital caliper every 5 days for 4-5 weeks.

Techniques: Stable Transfection

Journal: Oncotarget

Article Title: The tumor suppressive role of CAMK2N1 in castration-resistant prostate cancer

doi:

Figure Lengend Snippet: (A-J) Serial measurements were conducted every five days of DU145 and PC3 tumors stably expressing CAMK2N1, injected into nude mice. The data is shown as mean ± SEM for N>6 separate tumors for each group. (A, F) Images of tumors dissected out from the sacrificed mice. (B, G) The tumor size (mm3) versus days of post injection. (C, H) Reduction in tumor weight after resection at the end of experiment. (D-E, I-J) IHC staining detected the expression of CAMK2N1 in DU145 and PC3 tumor tissues of nude mice.

Article Snippet: 2 × 10 6 DU145 cells and PC3 cells expressing CAMK2N1 were implanted subcutaneously into 4-6-week-old athymic female nude mice purchased from Beijing HFK Bio-Technology.co., LTD. Tumor growth was measured using a digital caliper every 5 days for 4-5 weeks.

Techniques: Stable Transfection, Expressing, Injection, Immunohistochemistry

Journal: Oncotarget

Article Title: The tumor suppressive role of CAMK2N1 in castration-resistant prostate cancer

doi:

Figure Lengend Snippet: DU145 cells stably overexpressed CAMK2N1. These cells were analyzed for apoptosis by (A) Annexin V staining. Quantification of colony numbers and sizes were shown as mean ± SEM for N>5 separate experiments. (B) TUNEL staining was conducted to assess the effect of CAMK2N1 on cellular apoptosis in vivo . The percentage of apoptotic cells was increased in DU145 tumor tissues derived from nude mice. Quantification of TUNEL staining was shown as mean ± SEM for N=4 separate experiments. (C) IHC staining detected the expression of Bax, Bcl2, p21 and Ki67 in DU145 tumor tissue of nude mice. Overexpression of CAM2KN1 in DU145 and PC3 tumor tissue decreased Bcl-2, Ki67 protein expression and increased p21, Bax protein expression. Data for quantified IHC was shown as mean ± SEM for N = 4 tumors in each group.

Article Snippet: 2 × 10 6 DU145 cells and PC3 cells expressing CAMK2N1 were implanted subcutaneously into 4-6-week-old athymic female nude mice purchased from Beijing HFK Bio-Technology.co., LTD. Tumor growth was measured using a digital caliper every 5 days for 4-5 weeks.

Techniques: Stable Transfection, Staining, TUNEL Assay, In Vivo, Derivative Assay, Immunohistochemistry, Expressing, Over Expression

Journal: Oncotarget

Article Title: The tumor suppressive role of CAMK2N1 in castration-resistant prostate cancer

doi:

Figure Lengend Snippet: (A-B) Microarray gene expression analysis was conducted in DU145 cells stably overexpressing CAMK2N1 (>1.5-fold and p < 0.05). (A) Functional analysis of differentially expressed genes. Gene Ontology (GO) Biological Process (BP) terms were ranked by score (score > 1.3). (B) CAMK2N1 induced and repressed clusters of genes that were chosen from the pathways. (C-D) Expression levels of ErbB2, AKT, MEK1, ERK1/2, NFκβ, Bcl-2, BAX, and p21 were determined by Western blot in DU145 cells with stably overexpressing CAMK2N1. Similar changes as to those found in the microarray analysis were observed. Each figure represents three independent experiments. (E) DU145 tumors stably expressing CAMK2N1, and mRNA levels of ErbB2, Bcl-2, NF-κβ, AKT1, AR, p21, and Bax were determined by qRT-PCR. CAMK2N1 decreased ErbB2, BCL-2, NF-κβ, AKT1, AR mRNA levels and increased p21, Bax mRNA levels in DU145 tumor tissues. The data is shown as mean ± SEM for N=4 separate tumors for each group.

Article Snippet: 2 × 10 6 DU145 cells and PC3 cells expressing CAMK2N1 were implanted subcutaneously into 4-6-week-old athymic female nude mice purchased from Beijing HFK Bio-Technology.co., LTD. Tumor growth was measured using a digital caliper every 5 days for 4-5 weeks.

Techniques: Microarray, Gene Expression, Stable Transfection, Functional Assay, Expressing, Western Blot, Quantitative RT-PCR

Journal: Frontiers in Genetics

Article Title: Long Non-coding RNA SNHG17 Promotes Cell Proliferation and Invasion in Castration-Resistant Prostate Cancer by Targeting the miR-144/CD51 Axis

doi: 10.3389/fgene.2020.00274

Figure Lengend Snippet: SNHG17 expression was up-regulated and positively correlated with CD51 expression in CRPC tissues and cells. (A) The expression of SNHG17 in HSPC and CRPC tissues, as detected by RT-QPCR; (B) The mRNA expression of CD51 HSPC and CRPC tissues, as detected by RT-QPCR; (C) The association between CD51 and SNHG17 expression in HSPC tissues; (D) The association between CD51 and SNHG17 expression in CRPC tissues; (E) SNHG17 expression in HSPC cell line (LNCaP) and CRPC cell lines (PC-3 and C4-2), as detected by RT-QPCR; (F) The expression of CD51 in HSPC cell line (LNCaP) and CRPC cell lines (PC-3 and C4-2), as detected by RT-QPCR; Data expressed as the mean ± SE . * P < 0.05, ns, not significant. The representative results of 3 independent experiments are shown.

Article Snippet: Human prostate cancer cell lines LNCaP, C4-2, PC-3, and DU145 were purchased from GeneChem (Shanghai, China).

Techniques: Expressing, Quantitative RT-PCR

Journal: Frontiers in Oncology

Article Title: Enzalutamide-Induced Upregulation of PCAT6 Promotes Prostate Cancer Neuroendocrine Differentiation by Regulating miR-326/HNRNPA2B1 Axis

doi: 10.3389/fonc.2021.650054

Figure Lengend Snippet: PCAT6 expression was increased in NEPC cells and tissues. (A) qPCR analysis of the top 10 lncRNAs (PCAT6, LncRNA7816-1, LncRNA2323-2, LINC00319, lncRNA-p21, Malat1, PCA3, LncRNA8802, LncRNA1698-1 and LncRNA2303) mRNA expression in NCI-H660 cells and LNCaP cells. (B) qPCR analysis of PCAT6 mRNA expression in NE-like cells (PC3, DU145, and NCI-H660) and LNCaP and C4-2 cells. (C, D) The mRNA expression of PCAT6 was determined in LNCaP cells treated with different Enza concentration (2 μM, 4 μM, 8 μM, 10 μM) for 72 h or 10 μM for different times (0 h, 12 h, 24 h, 48 h, 72 h). (E) Fluorescent in situ hybridization (FISH) assay of the PCAT6 expression in LNCaP and NCI-H660 cells. (F) Fluorescent in situ hybridization (FISH) assay of the PCAT6 expression in LNCaP cells with 10 μM Enza treatment for 72 h. (G) qPCR analysis of PCAT6 expression in Pca tissues and adjacent normal tissues. (H) qPCR analysis of PCAT6 expression in NEPC (n=9) and CRPC (n=18) samples. * p <0.05. ** p <0.01.

Article Snippet: NE-like cells (PC3, DU145, and NCI-H660), LNCaP, C4-2 cells were purchased from ScienCell (Santiago, California, USA).

Techniques: Expressing, Concentration Assay, In Situ Hybridization

Journal: Frontiers in Oncology

Article Title: Enzalutamide-Induced Upregulation of PCAT6 Promotes Prostate Cancer Neuroendocrine Differentiation by Regulating miR-326/HNRNPA2B1 Axis

doi: 10.3389/fonc.2021.650054

Figure Lengend Snippet: PCAT6 promoted NED, proliferation, and invasion of PCa cells in vitro . (A) qPCR analysis of NE markers (NSE, SYP, and ChgA) mRNA expression in LNCaP cells after PCAT6 overexpression. (B, C) Western blot analysis for NSE, SYP, and ChgA protein level in LNCaP cells after PCAT6 overexpression. (D) qPCR analysis of NSE, SYP, and ChgA mRNA expression in NCI-H660 cells after PCAT6 knockdown. (E) LNCaP cell proliferation was assayed using CCK-8 after PCAT6 overexpression. (F) NCI-H660 cell proliferation was assayed using CCK-8 after PCAT6 knockdown. (G, H) LNCaP cell invasion was assessed using transwell invasion assay after PCAT6 overexpression. (I, J) NCI-H660 cell invasion was assessed using transwell invasion assay after PCAT6 knockdown. * p <0.05. ** p <0.01.

Article Snippet: NE-like cells (PC3, DU145, and NCI-H660), LNCaP, C4-2 cells were purchased from ScienCell (Santiago, California, USA).

Techniques: In Vitro, Expressing, Over Expression, Western Blot, CCK-8 Assay, Transwell Invasion Assay

Journal: Frontiers in Oncology

Article Title: Enzalutamide-Induced Upregulation of PCAT6 Promotes Prostate Cancer Neuroendocrine Differentiation by Regulating miR-326/HNRNPA2B1 Axis

doi: 10.3389/fonc.2021.650054

Figure Lengend Snippet: PCAT6 functioned as a ceRNA and sponged miR-326. (A) Starbase v2.0 database was used to predict potential miRNA binding sites in PCAT6. (B) qPCR analysis of miR-326 mRNA expression in NE-like cells (PC3, DU145, and NCI-H660) and LNCaP and C4-2 cells. (C) Schematic representation of the miR-326 site in PCAT6-3’UTR. (D) Luciferase activity was assayed in LNCaP cells co-transfected with miR-326 and luciferase reporters containing PCAT6-WT or PCAT6-Mut. Data are presented as the relative ratio of firefly luciferase activity to renilla luciferase activity. (E) Luciferase activity was assayed in LNCaP cells co-transfected with luciferase reporters containing PCAT6-WT and miR-326 or miR-326-Mut. (F) The direct binding of PCAT6 to miR-326 was affirmed using luciferase reporters in LNCaP cells treated with miR-326 or miR-326-mut. (G) qPCR analysis of miR-326 level in CRPC tissues (n=17) and NEPC tissues (n=9). ** p <0.01.

Article Snippet: NE-like cells (PC3, DU145, and NCI-H660), LNCaP, C4-2 cells were purchased from ScienCell (Santiago, California, USA).

Techniques: Binding Assay, Expressing, Luciferase, Activity Assay, Transfection