Lai, Pei-LunPei-LunLaiChen, Ting-ChunTing-ChunChenFeng, Chun-YenChun-YenFengLin, HsuanHsuanLinNg, Chi-HouChi-HouNgChen, YunYunChenHsiao, MichaelMichaelHsiaoLu, JeanJeanLuHSIAO-CHUN HUANG2020-12-292020-12-2920201792-1074https://www.scopus.com/inward/record.url?eid=2-s2.0-85090361007&partnerID=40&md5=a1aa21df66f726b17bcb3dddc3520e4bhttps://scholars.lib.ntu.edu.tw/handle/123456789/535685Colorectal cancer (CRC) is a leading cause of cancer-associated mortality worldwide; therefore, there is an emerging need for novel experimental models that allow for the identification and validation of biomarkers for CRC-specific progression. In the present study, a repeated sphere-forming assay was used as a strategy to select a malignant subpopulation from a CRC cell line, namely HCT116. The assay was validated by confirming that canonical stemness markers were upregulated in the sphere state at every generation of the selection assay. The resulting subpopulation, after eight rounds of selection, exhibited increased sphere-forming capacity in vitro and increased tumorigenicity in vivo. Furthermore, dipeptidase 1 (DPEP1) was identified as the major differentially expressed gene in the selected clone, and its depletion suppressed the elevated sphere-forming capacity in vitro and tumorigenicity in vivo. Overall, the present study established an experimental strategy to isolate a malignant subpopulation from a CRC cell line. Additionally, results from the present model revealed that DPEP1 may serve as a promising prognostic biomarker for CRC. ? 2020 Spandidos Publications. All rights reserved.Cell line; Colorectal cancer; DPEP1; Malignant subpopulation; Sphere-forming[SDGs]SDG3biological marker; complementary DNA; dipeptidase; dipeptidase 1; G protein coupled receptor kinase 5; glyceraldehyde 3 phosphate dehydrogenase; octamer transcription factor 4; transcription factor NANOG; unclassified drug; angiogenesis; animal experiment; Article; cancer prognosis; carcinogenicity; cell assay; cell culture; colorectal cancer; colorectal cancer cell line; down regulation; gene expression; HCT 116 cell line; human; human cell; image analysis; intestine epithelium; male; mouse; nonhuman; optical density; phase contrast microscopy; protein expression; radioimmunoprecipitation; real time reverse transcription polymerase chain reaction; RNA extraction; RNA sequencing; sphere derived adherent cell; sphere forming assay; tumor growth; tumor volume; upregulation; Western blottingjournal article10.3892/ol.2020.11829327826102-s2.0-85090361007WOS:000563938300092