Conditionally reprogrammed normal and primary tumor prostate epithelial cells: a novel patient-derived cell model for studies of human prostate cancer
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Olga A. Timofeeva1,2, Nancy Palechor-Ceron3, Guanglei Li4, Hang Yuan3, Ewa Krawczyk3, Xiaogang Zhong5, Geng Liu4, Geeta Upadhyay1, Aleksandra Dakic3, Songtao Yu3, Shuang Fang3, Sujata Choudhury3, Xueping Zhang1, Andrew Ju6, Myeong-Seon Lee7, Han C. Dan8, Youngmi Ji9, Yong Hou4, Yun-Ling Zheng1, Chris Albanese1,3, Johng Rhim9, Richard Schlegel3, Anatoly Dritschilo1,6, Xuefeng Liu3
1Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC, USA
2Department of Pathology and Laboratory Medicine, Lewis Katz Medical School, Temple University, Philadelphia, PA, USA
3Department of Pathology and Center for Cell Reprogramming, Georgetown University Medical Center, Washington, DC, USA
4Beijing Genome Research Institute, Shenzhen, Guangdong, China
5Department of Biostatistics, Bioinformatics, and Biomathematics, Georgetown University, Washington, DC, USA
6Department of Radiation Medicine, Georgetown University Medical School, Washington, DC, USA
7Department of Biomedical Science, Cheongju University, Sandang-Gu, Cheongju Shi Chungbuk, Republic of Korea
8Department of Pathology, University of Maryland, Baltimore, MD, USA
9Department of Surgery, Center for Prostate Disease Research, Uniformed Services University of the Health Sciences, Bethesda, MD, USA
Xuefeng Liu, email: [email protected]
Anatoly Dritschilo, email: [email protected]
Olga A. Timofeeva, email: [email protected]
Keywords: prostate cancer, conditional cell reprogramming, cell lines, patient-derived, cancer cell models
Received: June 13, 2016 Accepted: November 22, 2016 Published: December 21, 2016
Our previous study demonstrated that conditional reprogramming (CR) allows the establishment of patient-derived normal and tumor epithelial cell cultures from a variety of tissue types including breast, lung, colon and prostate. Using CR, we have established matched normal and tumor cultures, GUMC-29 and GUMC-30 respectively, from a patient’s prostatectomy specimen. These CR cells proliferate indefinitely in vitro and retain stable karyotypes. Most importantly, only tumor-derived CR cells (GUMC-30) produced tumors in xenografted SCID mice, demonstrating maintenance of the critical tumor phenotype. Characterization of cells with DNA fingerprinting demonstrated identical patterns in normal and tumor CR cells as well as in xenografted tumors. By flow cytometry, both normal and tumor CR cells expressed basal, luminal, and stem cell markers, with the majority of the normal and tumor CR cells expressing prostate basal cell markers, CD44 and Trop2, as well as luminal marker, CD13, suggesting a transit-amplifying phenotype. Consistent with this phenotype, real time RT-PCR analyses demonstrated that CR cells predominantly expressed high levels of basal cell markers (KRT5, KRT14 and p63), and low levels of luminal markers. When the CR tumor cells were injected into SCID mice, the expression of luminal markers (AR, NKX3.1) increased significantly, while basal cell markers dramatically decreased. These data suggest that CR cells maintain high levels of proliferation and low levels of differentiation in the presence of feeder cells and ROCK inhibitor, but undergo differentiation once injected into SCID mice. Genomic analyses, including SNP and INDEL, identified genes mutated in tumor cells, including components of apoptosis, cell attachment, and hypoxia pathways. The use of matched patient-derived cells provides a unique in vitro model for studies of early prostate cancer.
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