A convenient and effective strategy for the enrichment of tumor-initiating cell properties in prostate cancer cells.
Stem-like prostate cancer (PrCa) cells, also called PrCa stem cells (PrCSCs) or PrCa tumor-initiating cells (PrTICs), are considered to be involved in the mediation of tumor metastasis and may be responsible for the poor prognosis of PrCa patients. Currently, the methods for PrTIC sorting are mainly based on cell surface marker or side population (SP). However, the rarity of these sorted cells limits the investigation of the molecular mechanisms and therapeutic strategies targeting PrTICs. For PrTIC enrichment, we induced cancer stem cell (CSC) properties in PrCa cells by transducing three defined factors (OCT3/4, SOX2, and KLF4), followed by culture with conventional serum-containing medium. The CSC properties in the transduced cells were evaluated by proliferation, cell cycle, SP assay, drug sensitivity technology, in vivo tumorigenicity, and molecular marker analysis of PrCSCs compared with parental cells and spheroids. After culture with serum-containing medium for 8 days, the PrCa cells transduced with the three factors showed significantly enhanced CSC properties in terms of marker gene expression, sphere formation, chemoresistance to docetaxel, and tumorigenicity. The percentage of CD133+/CD44+cells was ninefold higher in the transduced cell population than in the adherent PC3 cell population (2.25 ± 0.62 vs. 0.25 ± 0.12 %, respectively), and the SP increased to 1.22 ± 0.18 % in the transduced cell population, but was undetectable in the adherent population. This method can be used to obtain abundant PrTIC material and enables a complete understanding of PrTIC biology and development of novel therapeutic agents targeting PrTICs.
Department of Urology, Zhujiang Hospital, Southern Medical University, No. 253 Gongyedadaozhong Road, Haizhu District, Guangzhou, 510282, China.
Yiming
Y
Zhang
Department of Urology, Zhujiang Hospital, Southern Medical University, No. 253 Gongyedadaozhong Road, Haizhu District, Guangzhou, 510282, China.
Yiqiang
Y
Huang
Department of Urology, Zhujiang Hospital, Southern Medical University, No. 253 Gongyedadaozhong Road, Haizhu District, Guangzhou, 510282, China.
Zhong
Z
Jin
Department of Urology, Zhujiang Hospital, Southern Medical University, No. 253 Gongyedadaozhong Road, Haizhu District, Guangzhou, 510282, China.
Xiezhao
X
Li
Department of Urology, Zhujiang Hospital, Southern Medical University, No. 253 Gongyedadaozhong Road, Haizhu District, Guangzhou, 510282, China.
Bingkun
B
Li
Department of Urology, Zhujiang Hospital, Southern Medical University, No. 253 Gongyedadaozhong Road, Haizhu District, Guangzhou, 510282, China.
Peng
P
Xu
Department of Urology, Zhujiang Hospital, Southern Medical University, No. 253 Gongyedadaozhong Road, Haizhu District, Guangzhou, 510282, China. professor_huangp@163.com.
Peng
P
Huang
Department of Urology, Zhujiang Hospital, Southern Medical University, No. 253 Gongyedadaozhong Road, Haizhu District, Guangzhou, 510282, China. dr_liuchunxiao@sina.com.
Chunxiao
C
Liu
D000071916:AC133 Antigen / Q000235:genetics / Q000378:metabolism; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2 / Q000235:genetics / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D002453:Cell Cycle / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000235:genetics; D002471:Cell Transformation, Neoplastic / Q000235:genetics / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D018960:Hyaluronan Receptors / Q000235:genetics / Q000378:metabolism; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics*; D008297:Male; D008819:Mice, Nude; D008856:Microscopy, Fluorescence; D014411:Neoplastic Stem Cells / Q000378:metabolism*; D050814:Octamer Transcription Factor-3 / Q000235:genetics*; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055748:SOXB1 Transcription Factors / Q000235:genetics*; D018874:Spheroids, Cellular / Q000378:metabolism; D014162:Transfection; D014183:Transplantation, Heterologous; D047368:Tumor Burden / Q000235:genetics
D000071916:AC133 Antigen; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018960:Hyaluronan Receptors; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors
Enrichment; Prostate cancer; Reprogramming; Tumor-initiating cells