Functional analysis of protein disulfide isomerase P5 in glioblastoma cells as a novel anticancer target

Horibe,Tomohisa; Torisawa,Aya; Masuda,Yoshie; Kawakami,Koji

doi:10.3892/or.2018.6868

Functional analysis of protein disulfide isomerase P5 in glioblastoma cells as a novel anticancer target

Authors:
- Tomohisa Horibe
- Aya Torisawa
- Yoshie Masuda
- Koji Kawakami
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Affiliations: Department of Pharmacoepidemiology, Graduate School of Medicine and Public Health, Kyoto University, Kyoto 606‑8501, Japan
Published online on: November 15, 2018 https://doi.org/10.3892/or.2018.6868
Pages: 961-972

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Abstract

P5, which is a member of the protein disulfide isomerase family, possesses isomerase and chaperone activity in vitro; however, the physiological functions of this enzyme in cells remain unclear. To understand the important roles of P5 in cancer cells, the present study examined its expression on the surface of normal and cancer cell lines by flow cytometry using an affinity‑purified anti‑P5 antibody labeled with 6‑(fluorescein‑5‑carboxamido) hexanoic acid succinimidyl ester. P5 expression was increased on the surface of various cancer cell lines, including leukemia cells, and glioblastoma, breast, colon, ovarian and uterine cervical cancer cells, compared with normal cells. However, P5 was constantly expressed within both normal and cancer cell lysates, and its total expression levels were not significantly different between the cells. P5 knockdown in glioblastoma cells by small interfering RNA affected Bip promoter activation during cancer cell growth, and significantly inhibited cancer cell growth and migration. Immunoprecipitation using an anti‑P5 antibody in cancer and normal cells demonstrated that vimentin was bound to P5, predominantly in U251 glioblastoma cells. P5 knockdown in glioblastoma cells did not affect the protein expression levels of vimentin; however, it did affect the expression of numerous epithelial‑mesenchymal transition markers, including Snail and Slug. These results suggested that P5 may serve an important role in cancer cell growth, and may be considered an attractive and potent target for the treatment of glioblastoma.

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