The lineage relationship between prostate adenocarcinoma and small cell carcinoma was studied by using the LuCaP family of xenografts established from primary neoplasm to metastasis. Expression of four stem cell transcription factor (TF) genes, LIN28A, NANOG, POU5F1, SOX2, were analyzed in the LuCaP lines. These genes, when force expressed in differentiated cells, can reprogram the recipients into stem‐like induced pluripotent stem (iPS) cells. Most LuCaP lines expressed POU5F1, while LuCaP 145.1, representative of small cell carcinoma, expressed all four. Through transcriptome database query, many small cell carcinoma genes were also found in stem cells. To test the hypothesis that prostate cancer progression from “differentiated” adenocarcinoma to “undifferentiated” small cell carcinoma could involve re‐expression of stem cell genes, the four TF genes were transduced via lentiviral vectors into five adenocarcinoma LuCaP lines—70CR, 73CR, 86.2, 92, 105CR—as done in iPS cell reprogramming. The resultant cells from these five transductions displayed a morphology of small size and dark appearing unlike the parentals. Transcriptome analysis of LuCaP 70CR* (“*” to denote transfected progeny) revealed a unique gene expression close to that of LuCaP 145.1. In a prostate principal components analysis space based on cell‐type transcriptomes, the different LuCaP transcriptome datapoints were aligned to suggest a possible ordered sequence of expression changes from the differentiated luminal‐like adenocarcinoma cell types to the less differentiated, more stem‐like small cell carcinoma types, and LuCaP 70CR*. Prostate cancer progression can thus be molecularly characterized by loss of differentiation with re‐expression of stem cell genes. J. Cell. Physiol. 231: 2040–2047, 2016. © 2016 Wiley Periodicals, Inc.

You have accessJournal of UrologyProstate Cancer: Basic Research & Pathophysiology I1 Apr 2016MP62-18 CONVERSION OF ADENOCARCINOMA TO SMALL CELL CARCINOMA-LIKE BY REPROGRAMMING Gisely Borges, Eneida Vencio, Sue-Ing Quek, Adeline Chen, Diego Salvanha, Ricardo Vencio, Holly Nguyen, Robert Vessella, Christopher Cavanaugh, Carol Ware, Pamela Troisch, and Alvin Liu Gisely BorgesGisely Borges More articles by this author , Eneida VencioEneida Vencio More articles by this author , Sue-Ing QuekSue-Ing Quek More articles by this author , Adeline ChenAdeline Chen More articles by this author , Diego SalvanhaDiego Salvanha More articles by this author , Ricardo VencioRicardo Vencio More articles by this author , Holly NguyenHolly Nguyen More articles by this author , Robert VessellaRobert Vessella More articles by this author , Christopher CavanaughChristopher Cavanaugh More articles by this author , Carol WareCarol Ware More articles by this author , Pamela TroischPamela Troisch More articles by this author , and Alvin LiuAlvin Liu More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.926AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Most prostate tumors are adenocarcinoma with glandular differentiation, and a small percentage, found most frequently in advanced diseases, are small cell neuroendocrine carcinoma without glandular differentiation. The LuCaP series is a family of prostate cancer xenografts propagated in male mice. LuCaP CR variants were obtained from passages in castrated hosts. Expression of stem cell transcription factor (TF) genes LIN28A, NANOG, POU5F1, SOX2 was examined in the LuCaP lines because some LuCaP lines were found to express stem cell markers. These four genes when forced expressed in somatic cells could induce reprogramming. METHODS The LuCaP cells were harvested and frozen by using a special protocol that preserved their viability. Gene expression was studied by RT-PCR. Adenocarcinoma LuCaP cells were plated on mouse embryonic fibroblast (MEF) feeder, which allowed them to grow in vitro. The cancer cells were transfected with lentiviral vectors containing the TF genes. Transfected cells on MEF were monitored over a period of 1 month. DNA microarrays were used to obtain cell type transcriptomes. RESULTS Most LuCaP lines expressed POU5F1, and LIN28A was upregulated in LuCaP 73CR vs. LuCaP 73. The small cell carcinoma LuCaP 145.1 expressed all four. Its expression of B2M (β-2 microglobulin) was also lower than that of other LuCaP lines. Low B2M expression is characteristic of stem cells. Many other LuCaP 145.1 genes (COTL1, ENO2, IGFBP2, ROBO1, chromogranin A) were also found in embryonic stem (ES) cells. TF transfection was carried out on five (AR+/PSA+) adenocarcinoma LuCaP lines: 70CR, 73CR, 86.2, 92, 105CR. All produced small dark cells as compared to large bright parental cells. The colony morphology was unlike that of ES cells, and more like that of PC3 cancer cell line, which was established from a bone metastasis of high-grade undifferentiated prostate carcinoma. The small cells started to appear around day 10 post-transfection and multiplied until individual colonies were visible at day 21. No such cells were seen in mock transfected. The transcriptome of LuCaP 70CR* (* to indicate transfected progeny) was very different from that of parental LuCaP 70CR as measured by Δ=91.72 in a prostate principal components analysis space. Gene expression of LuCaP 70CR* was closest to that of LuCaP 145.1 by Δ=45.85 (vs. Δ=78.04 with ES cells). CONCLUSIONS The adenocarcinoma phenotype could be converted to that of small cell carcinoma-like by stem cell TF genes (as in reprogramming). Cancer progression can be characterized molecularly as cellular de-differentiation from differentiated adenocarcinoma to undifferentiated stem-like small cell carcinoma. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e819 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Gisely Borges More articles by this author Eneida Vencio More articles by this author Sue-Ing Quek More articles by this author Adeline Chen More articles by this author Diego Salvanha More articles by this author Ricardo Vencio More articles by this author Holly Nguyen More articles by this author Robert Vessella More articles by this author Christopher Cavanaugh More articles by this author Carol Ware More articles by this author Pamela Troisch More articles by this author Alvin Liu More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

BACKGROUNDVoided urine samples have been shown to contain cells released from prostate tumors. Could good quality RNA from cells in urine be obtained from every donor for multimarker analysis? In addition, could urine donation be as simple as possible, a practical consideration for a lab test, without involving a prostate massage (as indicated for PCA3 testing), which precludes frequent collection; needing it done at a specific time of day (e.g., first or second urine); and requiring prompt processing of samples in clinics with limited molecular biology capability?METHODSCollected urine samples were pelleted, and the RNA isolated was processed for cDNA synthesis and in vitro transcription to generate amplified sense aRNA. The resultant aRNA was rigorously analyzed for possible introduced changes. DMSO was used as a cell preservative for frozen storage of urine samples.RESULTSGood quality aRNA was obtained for over 100 samples collected at two different institutions. The process of RNA amplification removed co-isolated DNA in some samples, which did not affect RNA amplification. Amplification did not amplify genes that were absent and produce other expression alterations. The sense aRNA could be used to generate urinary transcriptomes specific to individual patients. No chaotropic agents for RNA preservation were added to the urine samples so that the supernatant could be used for analysis of secreted protein biomarkers. The time of donation was not important since patients were seen during the entire day. DMSO was an effective cell preservative for freezing urine.CONCLUSIONSUrinary RNA can be readily isolated and amplified for prostate cancer biomarker analysis. Individual patients had unique set of transcripts derived from their tumor. Prostate 75:1886-1895, 2015. (c) 2015 Wiley Periodicals, Inc.