Journal: Nature Communications

Article Title: OXPHOS remodeling in high-grade prostate cancer involves mtDNA mutations and increased succinate oxidation

doi: 10.1038/s41467-020-15237-5

Figure Lengend Snippet: a Relative GM-OXPHOS respiratory capacity with glutamate&malate (GM P /NS E ) in benign (blue) or malignant (red) samples carrying either no or only synonymous HPs (−) ( N BE = 36 and N CA = 23) versus samples carrying non-synonymous HPs (+) ( N BE = 14 and N CA = 27) in the coding regions of the mt-genome. Values present mean ± SD. Differences in mean values were tested for significance using Wilcoxon rank-sum test. b Comparison of relative GM-OXPHOS capacity in samples without mutations (−) ( N BE = 38 and N CA = 37) versus samples with mutations (+) ( N BE = 12 and N CA = 13) within the non-coding (control) region of the mt-genome. Values represent mean ± SD. c Impact of location of non-synonymous HPs on relative GM-OXPHOS capacities. Tumor tissue samples were categorized according to no HPs (−) (orange; N = 24), and HPs located in genes encoding proteins of CIII–CV (CO, ATP, CYB; green; N = 10) or in genes encoding proteins of CI (ND1–ND5; blue; N = 20). d – e Relative GM-OXPHOS (GM P /NS E , d ) and relative S-ET (S E /NS E , e ) respiratory capacities in malignant tissue samples harboring non-synonymous HPs in CI-coding mt-genes. Tumors were grouped into samples carrying no non-synonymous HPs ((−); N = 23), samples with variant levels of 30–60% (30–60%; N = 6) and samples with variant levels >60% (>60%; N = 4). Differences in mean values were tested for significance using one-way ANOVA followed by Tukey’s HSD test. f N-pathway (blue) and S-pathway (orange) respiratory capacities in malignant samples harboring high-level (>60%) non-synonymous HPs in either CIV-coding genes ( N = 4) or CI-coding genes ( N = 4). Differences of in mean values were tested for significance using Wilcoxon rank-sum test. Values presented in c-f represent mean values and individual data points. g S-pathway OXPHOS capacity upregulation by partial inhibition of N-pathway oxidative flux in benign (RWPE1, N = 3; EP156T, N = 3) and malignant (PC3, N = 6; LNCaP, N = 4; DuCaP, N = 3, N represents number of biologically independent experiments) prostate cell lines. Relative S-pathway OXPHOS capacity (normalized to total respiratory capacity, NS E ) with different degrees of N-pathway inhibition is shown for the five cell lines. Values represent mean ± SD. Source data are provided as a Source Data file.

Article Snippet: DuCaP prostate cancer cells were a gift from Dr. Schalken (Radboud University, Nijmegen, The Netherlands).

Techniques: Comparison, Control, Variant Assay, Inhibition

Journal: Cell Death & Disease

Article Title: Integrated analysis reveals FOXA1 and Ku70/Ku80 as targets of ivermectin in prostate cancer

doi: 10.1038/s41419-022-05182-0

Figure Lengend Snippet: Cell viability was measured by the MTT assay in AR-positive cells (LNCaP, C4-2, and 22RV1 ( A ), AR-negative cells (DU145 and PC-3) ( B ), and BPH-1 or prostate primary cells from BPH patients ( C ) treated with the indicated concentrations of ivermectin for either 24, 48, or 72 h.

Article Snippet: Prostate cancer cell lines LNCaP, VCaP, and 22RV1 were purchased from Procell Life Science & Technology Co. Ltd. (Wuhan, China).

Techniques: MTT Assay

Journal: Cell Death & Disease

Article Title: Integrated analysis reveals FOXA1 and Ku70/Ku80 as targets of ivermectin in prostate cancer

doi: 10.1038/s41419-022-05182-0

Figure Lengend Snippet: A The ivermectin arrest cell cycle at G0/G1 was measured by flow cytometry. LNCaP, C4-2, and 22RV1 cells were treated with ivermectin at 4, 8, and 12 μM for 48 h. B Ivermectin induced cell apoptosis detected by PI/Annexin V staining. Cells were treated as in A. The PI + /Annexin V + and PI-/Annexin V + cells were calculated as apoptotic cells. C Western blot analysis of PARP and cleaved-Caspase-3 (c-Caspase-3) in cells treated with ivermectin for 48 h. D Ivermectin increased DNA damage. DNA fragments were shown as comet images in alkaline gel electrophoresis. The tail moment was used to quantify the DNA damage in the treatment of ivermectin for 48 h. E Western blot analysis of γH2A.X in cells treated with the ivermectin for 48 h. F Tumor volume of 22RV1 xenografts after castration treated with vehicle (con) or ivermectin (10 mg/kg, n = 5 for each group). G Representative images of Ki67, γH2A.X, and PSA immunostaining, in 22RV1 tumors treated with vehicle or ivermectin.

Article Snippet: Prostate cancer cell lines LNCaP, VCaP, and 22RV1 were purchased from Procell Life Science & Technology Co. Ltd. (Wuhan, China).

Techniques: Flow Cytometry, Staining, Western Blot, Nucleic Acid Electrophoresis, Immunostaining

Journal: Cell Death & Disease

Article Title: Integrated analysis reveals FOXA1 and Ku70/Ku80 as targets of ivermectin in prostate cancer

doi: 10.1038/s41419-022-05182-0

Figure Lengend Snippet: A Western blot analysis of AR and PSA in LNCaP and C4-2 cells treated with ivermectin for 48 h. B RT-qPCR analysis of AR target genes ( KLK3 , TMPRSS2 , and NKX3 -1) in LNCaP and C4-2 cells treated with ivermectin for 48 h. C Western blot analysis of FL-AR, ARVs, PSA, and UBE2C in ivermectin-treated 22RV1 cells at 48 h. D RT-qPCR analysis of KLK3 and ARV target genes ( UBE2C and CDC20 ) in 22RV1 cells treated with ivermectin for 48 h. E Western blot analysis of FL-AR, ARVs, PSA, PARP, and γH2A.X in the other two ARV-positive cells lines, LN95 and VCaP, treated with ivermectin for 48 h. F Western blot analysis of AR, PSA, PARP, and γH2A.X in LNCaP and C4-2 cells after the implementation of 4 μM and 8 μM of ivermectin with or without 1 nM R1881. G Ivermectin inhibited the cell cycle at G0/G1 in the presence of R1881. LNCaP and C4-2 cells were treated with ivermectin at 4 and 8 μM for 48 h in the absence or presence of 1 nM R1881. H Cell viability was measured by the MTT assay. LNCaP and C4-2 cells were treated with indicated concentrations of ivermectin for 48 h with or without 5 μM and 10 μM enzalutamide for 48 h.

Article Snippet: Prostate cancer cell lines LNCaP, VCaP, and 22RV1 were purchased from Procell Life Science & Technology Co. Ltd. (Wuhan, China).

Techniques: Western Blot, Quantitative RT-PCR, MTT Assay

Journal: Cell Death & Disease

Article Title: Integrated analysis reveals FOXA1 and Ku70/Ku80 as targets of ivermectin in prostate cancer

doi: 10.1038/s41419-022-05182-0

Figure Lengend Snippet: A GSEA showed that genes induced by FOXA1 were inhibited by ivermectin in C4-2 cells. B RT-qPCR analysis of FOXA1 induced genes ( CDKN3 , CDCA2 , and CAMKK2 ) and FOXA1 repressed EMT-associated genes ( MET , MMP7 , and SOX9 ) in C4-2 cells treated with ivermectin for 48 h. C Western blot analysis of FOXA1 and N-cadherin in LNCaP and C4-2 cells treated with ivermectin for 48 h. D ChIP–qPCR analysis for FOXA1 or AR occupancy, and FAIRE–qPCR analysis of chromatin accessibility at a target regulated by AR and FOXA1 (KLK3 and NKX3-1) in C4-2 cells treated with ivermectin. E ChIP–qPCR analysis for FOXA1 and FAIRE-PCR analysis of chromatin accessibility at a target regulated by FOXA1 (E2F1 and MET) in C4-2 cells treated with ivermectin. F FOXA1 knockdown impaired the ivermectin-repressed expression of KLK3 and E2F1 genes. mRNA levels were measured 48 h after the implementation of the ivermectin treatment and siRNA transfection by RT-qPCR in C4-2 cells. G , H Western blots showing thermostable FOXA1 and AR following indicated heat shocks in the presence (+) or absence (−) of 50 μM ivermectin in LNCaP ( G ) and C4-2 ( H ) cells. I Western blots showing thermostable FOXA1 following indicated heat shocks in the presence (+) or absence (−) of 50 μM ivermectin in 22RV1 cells. J GSEA showed the inactivation of FOXA1 induced genes in 22RV1 cells after the ivermectin treatment. K RT-qPCR analysis of FL-AR and ARv7 in 22RV1 cells treated with ivermectin for 48 h. L ChIP–qPCR analysis for FOXA1 and FAIRE–qPCR analysis of chromatin accessibility at KLK3 and E2F1 in 22RV1 cells treated with ivermectin.

Article Snippet: Prostate cancer cell lines LNCaP, VCaP, and 22RV1 were purchased from Procell Life Science & Technology Co. Ltd. (Wuhan, China).

Techniques: Quantitative RT-PCR, Western Blot, ChIP-qPCR, Knockdown, Expressing, Transfection

Journal: Cell Death & Disease

Article Title: Integrated analysis reveals FOXA1 and Ku70/Ku80 as targets of ivermectin in prostate cancer

doi: 10.1038/s41419-022-05182-0

Figure Lengend Snippet: A Volcano plot of melting point difference calculated from the ivermectin versus DMSO controls in living 22RV1 cells. Blue circles represent significant melting temperature differences and red circles show all remaining proteins. B KEGG and GO pathways by KOBAS showed the enrichment pathway of the proteins with the melting temperature difference (ΔTm) more than ±3 °C. C Melting curves for Ku70/Ku80 generated from mass spectrum in 22RV1 cells. D , E Western blots showing thermostable Ku70/Ku80 following indicated heat shocks in the presence (+) or absence (−) of 50 μM ivermectin in LNCaP ( D ) and C4-2 ( E ) cells.

Article Snippet: Prostate cancer cell lines LNCaP, VCaP, and 22RV1 were purchased from Procell Life Science & Technology Co. Ltd. (Wuhan, China).

Techniques: Generated, Western Blot

Journal: eLife

Article Title: circFL-seq reveals full-length circular RNAs with rolling circular reverse transcription and nanopore sequencing

doi: 10.7554/eLife.69457

Figure Lengend Snippet:

Article Snippet: Cell line ( H. sapiens ) , VCaP , iCell Bioscience , RRID: CVCL_2235 , .

Techniques: