Abstract PO-138: The first Caribbean cell line, a prostate cancer cell line: ACRJ-PC28
Authors: Valentine Henkel , Aiken William , Morrison Belinda , Zhao Ziran , Fowle Holly , Wasserman Jason S. , Thompson Elon , Chin Warren , Young Mark , Clarke Shannique , Gibbs Denise , Harrison Sharon , McLaughlin Wayne , Kwok Tim , Jin Fang , Campbell Kerry , Horvath Anelia , Thompson Rory , Lee Norman , Zhou Yan , Graña Xavier , Ragin Camille , Badal Simone 
Published: January 2022

Abstract
Background Prostate cancer cell lines from diverse backgrounds are believed to hold the key to addressing current disparities in prostate cancer incidence and mortality rates among Black men. Despite decades of established cell line protocols, developing new cell lines, especially from the prostate, continues to attract less than ideal success rates (&lt;10%). Methods ACRJ-PC28 was developed from a transrectal needle biopsy (TRNB) that was mechanically dissociated and grown on 3T3 STO fibroblast feeder cells followed by establishment via inactivation of the CDKN2A locus and simultaneous expression of human telomerase. Characterization assays were carried out using growth curve analysis, immunoblots, immunohistochemistry, 3D cultures, immunofluorescence imaging, confocal microscopy, WGS and, RNA-Seq. Results ACRJ-PC28 has been passaged more than 40 times in vitro over 10 months with a doubling time of 45 hours. STR profiling confirms the novelty and human origin of the cell line while western blots indicate the cell line is of basal-luminal type; expresses p53, pRB and is AR negative. Soft agar anchorage-independent analysis indicated that the cells are transformed, confirmed by PCA analysis where ACRJ-PC28 cells cluster alongside other PCa tumour tissues. WGS confirms the absence of exonic mutations and the presence of intronic variants that seem to not affect function of AR, p53 and pRB. Meanwhile, RNA-Seq confirmed the expression of several neuroendocrine markers, most relative to the PC-3 cell line and 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. Cytotoxic assay indicated that a Cannabis extract was 3 times more potent in reducing the viability of ACRJ-PC28 compared to PC-3 cells. Conclusions This is the first cell line from the Caribbean and the novel methodology used in its establishment should help develop additional prostate cancer cell lines from the region in hopes of addressing the disparity in PCa among Black men.
Citation Format: Henkel Valentine, William Aiken, Belinda Morrison, Ziran Zhao, Holly Fowle, Jason S. Wasserman, Elon Thompson, Warren Chin, Mark Young, Shannique Clarke, Denise Gibbs, Sharon Harrison, Wayne McLaughlin, Tim Kwok, Fang Jin, Kerry Campbell, Anelia Horvath, Rory Thompson, Norman Lee, Yan Zhou, Xavier Graña, Camille Ragin, Simone Badal. The first Caribbean cell line, a prostate cancer cell line: ACRJ-PC28 [abstract]. In: Proceedings of the AACR Virtual Conference: 14th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2021 Oct 6-8. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2022;31(1 Suppl):Abstract nr PO-138.