Hepatocyte selection medium-enriched hepatocellular carcinoma cells are positive for α-fetoprotein and CD44

Tomizawa,Minoru; Shinozaki,Fuminobu; Motoyoshi,Yasufumi; Sugiyama,Takao; Yamamoto,Shigenori; Ishige,Naoki

doi:10.3892/ol.2017.6239

Hepatocyte selection medium-enriched hepatocellular carcinoma cells are positive for α-fetoprotein and CD44

Authors:
- Minoru Tomizawa
- Fuminobu Shinozaki
- Yasufumi Motoyoshi
- Takao Sugiyama
- Shigenori Yamamoto
- Naoki Ishige
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Affiliations: Department of Gastroenterology, National Hospital Organization, Shimoshizu Hospital, Yotsukaido, Chiba 284‑0003, Japan, Department of Radiology, National Hospital Organization, Shimoshizu Hospital, Yotsukaido, Chiba 284‑0003, Japan, Department of Neurology, National Hospital Organization, Shimoshizu Hospital, Yotsukaido, Chiba 284‑0003, Japan, Department of Rheumatology, National Hospital Organization, Shimoshizu Hospital, Yotsukaido, Chiba 284‑0003, Japan, Department of Pediatrics, National Hospital Organization, Shimoshizu Hospital, Yotsukaido, Chiba 284‑0003, Japan, Department of Neurosurgery, National Hospital Organization, Shimoshizu Hospital, Yotsukaido, Chiba 284‑0003, Japan
Published online on: May 24, 2017 https://doi.org/10.3892/ol.2017.6239
Pages: 899-902

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Abstract

Tissues surrounding hepatocellular carcinomas (HCCs) lack glucose. Hepatocyte selection medium (HSM) is deficient in glucose and is supplemented with galactose. HCC cells were cultured in HSM to investigate the stem cell markers α‑fetoprotein (AFP) and cluster of differentiation 44 (CD44). HCC cells (HLF and PLC/PRF/5 cells) were cultured in HSM. Viable cell numbers were determined on days 0 and 7 following culture in HSM. RNA was isolated and subjected to reverse transcription‑quantitative PCR (RT‑qPCR) to analyze the mRNA expression levels of AFP and CD44. Immunostaining was performed to analyze the protein levels of AFP and CD44. The number of viable cells was significantly decreased on day 7 following culture in HSM. The expression levels of AFP and CD44 increased on day 7 as assessed using RT‑qPCR. Immunostaining confirmed the results of RT‑qPCR analysis. The number of viable HCC cells was decreased in HSM, whereas the expression levels of AFP and CD44 increased. Therefore, HSM is potentially useful for the enrichment of HCC cells with cancer stem cell characteristics.

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