Prostate cancer cell lines from diverse backgrounds are important to addressing disparities in prostate cancer 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, three-dimensional cultures, immunofluorescence imaging, confocal microscopy, flow cytometry, whole-genome sequencing (WGS), and RNA sequencing (RNA-seq). ACRJ-PC28 has been passaged more than 40 times in vitro over 10 months with a doubling time of 45 hours. Short tandem repeat 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 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 androgen receptor (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 where ACRJ-PC28 cells cluster alongside other prostate cancer tumor tissues, yet was distinct. The novel methodology described should advance prostate cell line development, addressing the disparity in prostate cancer among Black men.Significance: Cell line development continues to attract less than 10% success rate. More than 98% of prostate cancer cell lines are from White men. This may contribute to the poorer response by Black men with prostate cancer to therapy compared with White men with prostate cancer, increasing overall survivorship among White men. The methodology described here to develop ACRJ-PC28, should advance the presence of Black prostate cancer cell lines thereby addressing prostate cancer disparity.

HNSCC is the sixth most common cancer worldwide and the seventh most common in Jamaica. Mortality rates for HNSCC in the Caribbean are among the highest in the world. Despite this there is no information on the molecular characterization of these cancers and the relative contribution of tobacco and HPV in this population. This study will examine the molecular features of HNSCC as it relates to p16 expression and HPV in patients from Jamaica. There were no previous published studies of this kind conducted in the English-speaking Caribbean.Methods: This retrospective cohort study was conducted using archival formalin-fixed paraffin-embedded tissue from patients diagnosed with HNSCC at the Department of Pathology, University of the West Indies and the National Public Health Laboratory from 2001–2010. Clinical information was extracted for all patients. P16 immunohistochemistry was performed using P16INK4A clone E6H4 mouse anti-human protein and identification of HR-HPV DNA was done by standard PCR using MY09/MY11 consensus primers of the L1 HR-HPV gene.Results: The study population consisted of 327 HNSCC cases with ages ranging from 18 to 92 years (mean, 65 yrs. ± 15 SD). There were 254 males and 63 females (10 unreported sex) with M: F ratio of 4:1. Most tumors were either moderately differentiated 147/327 (45%) or were poorly differentiated 55/327 (17%). This study cohort consisted of patients diagnosed with cancer of the larynx (57.5%), oral cavity (19.3%), other sites including lip, salivary gland and nasopharynx (14.7%) and oropharynx (8.6%). FFPE tissues from all cases were tested for p16; the majority 255/327 (78%) were negative and 71/327 (21.7%) were positive. All p16 + cases along with a subset of 60 randomly selected p16- cases were tested for HR-HPV by PCR. There were nine p16 + cases which were also positive for HR-HPV DNA. HPV 16 was detected in all cases, one case had HPV16 and 18, and another had HPV 16, 18, 39, 68 and 73. None of the sixty p16- cases were positive for HR-HPV. The nine patients that were HR-HPV + and p16 + had a mean age of 61 years ± 15 SD (range 39–83 years) with a M: F ratio of 8:1. All of these patients had moderately differentiated tumors, from oropharynx and larynx (44.4%) each and oral cavity (11.1%).Conclusion: In Jamaica, cancer of the larynx is the most common head and neck cancer. The prevalence of HPV-related HNSCC appears to be low with most patients with disease in non-oropharyngeal cancer sites. Based on what is known from the literature, our study results suggest that in Jamaica most patients are diagnosed with alcohol and tobacco driven disease. Interventions to reduce smoking and limit alcohol use are highly relevant in Jamaica. The role of HPV in HNSCC warrants further investigation in this population.

All Exonic Variants which are present in ACRJ-PC28. SIFT, Polyphen2, LRT, MutationTaster, MutationAssessor, and CADD were used to predict variant effect. Pop_Freq_max is the maximum population frequency of the variant in 1000Genome Project (Aug 2015), Exome Sequence Project (ESP6500) and Genome Aggregation Database (gnomAD). Total_Damaging represents number of times the variant was predicted as damaging by the 5 methods used. There are 605 variants that are predicted as damaging for 5 methods (green) while 296 variants are predicted as damaging for 4 out of 5 methods (orange-brown), among them is the variant in ATM (rs147934285) (dk brown). 3). Additionally, 539 variants are predicted as damaging for 3 out of 5 methods (blue-purple). The data generated and/or analyzed during the current study are available from the corresponding author on reasonable request.

Cell viability was determined using the MTS assay and OD readings were taken a 409nm. Plots were generated in Microsoft Excel and each data point represents the average of three replicates. Experiments shown are representative of two independent experiments.

Cell viability was determined using the MTS assay and OD readings were taken a 409nm. Plots were generated in Microsoft Excel and each data point represents the average of three replicates. Experiments shown are representative of two independent experiments.