American Association for Cancer Research
00085472can131560-sup-supp_data_3.xls (2.86 MB)

Supplementary Data 3 from Generation of Prostate Tumor–Initiating Cells Is Associated with Elevation of Reactive Oxygen Species and IL-6/STAT3 Signaling

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posted on 2023-03-30, 22:04 authored by Yi Qu, Anne Margrete Oyan, Runhui Liu, Yaping Hua, Jigang Zhang, Randi Hovland, Mihaela Popa, Xiaojun Liu, Karl A. Brokstad, Ronald Simon, Anders Molven, Biaoyang Lin, Wei-dong Zhang, Emmet McCormack, Karl-Henning Kalland, Xi-Song Ke

XLS file 2928K, Changed gene expression in D5HS comparing to EPT2-D5 cells



How prostate cancer is initiated remains a topic of debate. In an effort to establish a human model of prostate carcinogenesis, we adapted premalignant human prostate EPT2-D5 cells to protein-free medium to generate numerous tight prostate spheres (D5HS) in monolayer culture. In contrast to EPT2-D5 cells, the newly generated D5HS efficiently formed large subcutaneous tumors and subsequent metastases in vivo, showing the tumorigenicity of D5HS spheres. A striking production of interleukin (IL)-6 mRNA and protein was found in D5HS cells. The essential roles of IL-6 and the downstream STAT3 signaling in D5HS tumor sphere formation were confirmed by neutralizing antibody, chemical inhibitors, and fluorescent pathway reporter. In addition, elevated reactive oxygen species (ROS) produced upon protein depletion was required for the activation of IL-6/STAT3 in D5HS. Importantly, a positive feedback loop was found between ROS and IL-6 during tumor sphere formation. The association of ROS/IL-6/STAT3 to the carcinogenesis of human prostate cells was further examined in xenograft tumors and verified by limiting dilution implantations. Collectively, we have for the first time established human prostate tumor–initiating cells based on physiologic adaption. The intrinsic association of ROS and IL-6/STAT3 signaling in human prostate carcinogenesis shed new light on this relationship and define therapeutic targets in this setting. Cancer Res; 73(23); 7090–100. ©2013 AACR.