Project description:Identifying biological change from hormone-naive prostate cancer to CRPC is a major clinical challenge for developing therapeutic agents. Although the pathways that lead to CRPC are not fully understood, recent evidence demonstrates that androgen signaling is often maintained through varied mechanisms. Here, we investigated PCa tissues at each stage of progression from benign prostatic hyperplasia (BPH) to CRPC based on quantitative proteomic technology, including tissues after ADT therapy. MS-based quantitative proteomics approach based on 6-plex TMT (126-131) was performed in patient tissues from T2G2 to CRPC, and benign prostatic hyperplasia (BPH) patient tissues were used as a control. We analyzed the peptide samples using two types of high resolution and accuracy mass spectrometers as LTQ orbitrap velos and Q-exactive mass spectrometer. In total, 4,768 proteins were identified in this study, among which 4,069 proteins were quantified in the combined prostate cancer tissues. Among the quantified proteins, DEPs were 865 (21.2%), those with a quantitative ratio greater than 2 were considered as upregulated, whereas those with a quantitative ratio of less than 0.5 as downregulated. Based on quantitative protein results, we performed systematic bioinformatics analysis including GO, Interpro, KEGG pathway, functional enrichment-based cluster analysis on DEPs. Finally, we found that 15 proteins including FOXA1 and HMGN1-3 between T3G3, T3GX, and CRPC were increased despite ADT treatment. Among all target, we verified increased level of FOXA1 and HMGN1-3 in CRPC by immunoblotting and indirect ELISA. In summary, we provides intracellular mechanical changes on PCa tissues according to treatment before and after ADT by mean of regulating ADT treatment. In addition, this results were identified through bioinformatics analysis, and those were suggested as potential CRPC-related factors.
Project description:To investigate if there is a difference of N6-methyladenosine(m6A) modification between castration-resistant prostate cancer (CRPC) and castration-sensitive prostate cancer (CSPC), we collected 30 specimens, including 15 CRPC and 15 CSPC, to perform RNA-seq and MeRIP-seq. All specimens were postoperative tissues and each 5 CRPC or CSPC specimens were mixed into 1 sample to meet the RNA dosage of RNA-seq and MeRIP-seq.
Project description:Our studies revealed a novel oncogenic function of LSD1 in driving PCa progression by activating MYC signaling and mediating CRPC SEs activities, cotargeting LSD1 and BRD4 achieved significant synergistic effects in repressing CRPC tumor growth
Project description:The zebrafish is a powerful model for the study of hematopoietic stem and progenitor cells (HSPC). We have developed a novel HSPC-specific transgenic line (Runx1+23:GFP). We have used this line in time-lapse live imaging studies to track the migration of HSPC during development. We have also performed a chemical genetic screen to find small molecules that modulate HSPC numbers during development. Treating embryos from 2-3 days post fertilization (2-3 dpf) then fixing for in situ staining with HSPC probes cmyb and runx1, we found the compound lycorine increased HSPC numbers. Applying this compound during time-lapse live imaging showed increased accumulation of Runx+ HSPC in the caudal hematopoietic tissue (CHT). Treatment from 2-3 dpf, then washing off the compound, had a sustained effect on the size of the HSPC with Runx+ numbers higher at 5 and 7 dpf. We have performed microarray analysis to elucidate the molecular changes within HSPC and endothelial cells after Lycorine treatment. We treated Runx1+23:GFP;kdrl:DsRed2 embryos from 2-3 dpf with 75 uM lycorine in 1% DMSO. We then dissociated the embryos and sorted the Runx+ GFP cells, the kdrl+ DsRed2 cells, and the non-fluorescent negative cells from the total embryo as a comparator population. Total RNA was amplified and biotin labled for hybridization on Affymetrix microarrays. 18 samples were collected and analyzed. There are 3 biological replicates. There are 3 cell type populations: 1) Runx+ HSPC; 2) kdrl+ endothelial cells; 3) non-fluorescent negative cells. There are cell populations from dissociated Lycorine-treated embryo pools, and control DMSO-treated embryo pools.
Project description:We demonstrated to examine the effect of AhR antagonists on the specific lineage-biased differentiation of HSPC. To confirm this, CD34+ cells isolated from cord blood were cultured with two antagonists (CH223191 and StemRegenin 1) for 14 days to monitor their phenotype, and chromatin immunoprecipitation DNA was extracted from cultured cells on the 14th day of culture. We sought to explain the relationship between the action of AhR-antagonist complex and the megakaryocyte lineage-biased differentiation of HSPC with these ChIP-seq results.