Project description:Prostate cell lines from diverse backgrounds are important to addressing disparities in prostate cancer (PCa) incidence and mortality rates among Black men. ACRJ-PC28 was developed from a transrectal needle biopsy and established via inactivation of the CDKN2A locus and simultaneous expression of human telomerase. Characterization assays included growth curve analysis, immunoblots, IHC, 3D cultures, immunofluorescence imaging, confocal microscopy, flow cytometry, WGS and, RNA-Seq. RNA-Seq confirmed the expression of prostate specific genes alpha-methylacyl-CoA racemase (AMACR) and NKX3.1 and Neuroendocrine specific markers, synaptophysin (SYP) and enolase 2 (ENO2) and IHC confirmed the presence of AMACR. WGS confirms the absence of exonic mutations and the presence of intronic variants that appear to not affect function of AR, p53 and pRB. RNA-Seq data revealed numerous TP53 and RB1 mRNA splice variants and the lack of AR mRNA expression. The novel cell line described here should advance research addressing the disparity in prostate cancer among Black men.
Project description:Prostate cancer cell lines are particularly racially and clinically homogenous, and while there has been significant work in the development of additional models, few have been created without oncogenic transformation. Here we describe the derivation of a spontaneously immortalized primary prostate cancer cell line from a Black Caribbean patient with Gleason 7 prostate cancer – designated CaB34 - which presents a novel model for pre-clinical study of localized prostate cancer. From CaB34, a paired radioresistant subline CaB34-CF was generated using a clinically relevant fractionated radiotherapy schedule. Both CaB34 and CaB34-CF express prostate-specific markers including CK18, NKX3.1, and AMACR. Multi-omic analyses using RNAseq and shotgun proteomics identified NNMT as the most significantly dysregulated target in CaB34-CF. Bioinformatic analysis determined that NNMT was more abundant within prostate tumors compared to benign prostate, supporting a role in tumor progression. Knockdown studies of NNMT demonstrated significant radiosensitization of both CaB34-CF cells, which was largely a result of increased radiation-induced cellular senescence. Our findings present NNMT as a potential therapeutic target for sensitization of radioresistant disease. Together, our research provides a paired model of treatment naive and radioresistant disease to address mechanisms of therapy resistance, while expanding the racial diversity of prostate cancer cell lines for the research community.
Project description:This SuperSeries is composed of the following subset Series: GSE27022: Microarray studies of darkness stress and bleaching in the Caribbean coral Acropora palmata GSE27024: Microarray studies of darkness stress and bleaching in the Caribbean coral Montastraea faveolata Refer to individual Series
Project description:Prostate cell lines from diverse backgrounds are important to addressing disparities in prostate cancer (PCa) incidence and mortality rates among Black men. ACRJ-PC28 was developed from a transrectal needle biopsy and established via inactivation of the CDKN2A locus and simultaneous expression of human telomerase. Characterization assays included growth curve analysis, immunoblots, IHC, 3D cultures, immunofluorescence imaging, confocal microscopy, flow cytometry, WGS, and RNA-Seq. ACRJ-PC28 has been passaged more than 40 times in vitro over 10 months with a doubling time of 45 hours. STR profiling confirmed the novelty and human origin of the cell line. RNA-Seq confirmed the expression of prostate specific genes alpha-methylacyl-CoA racemase (AMACR) and NKX3.1 and Neuroendocrine specific markers synaptophysin (SYP) and enolase 2 (ENO2) and IHC confirmed the presence of AMACR. Immunoblots indicated the cell line is of basal-luminal type; expresses p53 and pRB and is AR negative. WGS confirmed the absence of exonic mutations and the presence of intronic variants that appear to not affect function of AR, p53, and pRB. RNA-Seq data revealed numerous TP53 and RB1 mRNA splice variants and the lack of AR mRNA expression. This is consistent with retention of p53 function in response to DNA damage and pRB function in response to contact inhibition. Soft agar anchorage-independent analysis indicated that the cells are transformed, confirmed by principal component analysis (PCA) where ACRJ-PC28 cells cluster alongside other PCa tumor tissues, yet was distinct. The novel methodology described should advance prostate cell line development, addressing the disparity in PCa among Black men.
Project description:The National Institute of Neurological Disorders and Stroke (NINDS) Human Genetics Resource Center: DNA and Cell Line Repository (the NINDS Repository): Cerebrovascular Disease/Stroke Study