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Inhibition of the PI3K-Akt signaling pathway enhances the sensitivity of Fas-mediated apoptosis in human gastric carcinoma cell line, MKN-45

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Abstract

It is well known that Fas ligand and anti-Fas antibodies can induce apoptosis, although some cancer cells are resistant to their stimuli. On the other hand, phosphatidylinositol 3’-kinase (PI3 K) and Akt mediate the survival signal and allow the cells to escape from apoptosis in various human cancers. Thus, we postulated that LY294002, a PI3 K inhibitor, should inactivate Akt, consequently inhibiting cell proliferation and increase apoptosis in the human gastric carcinoma cell line, MKN-45. Previously, we reported that MKN-45 was resistant against the anti-Fas antibody, CH-11, without interferon-gamma pretreatment in vitro. LY294002 caused a decrease of phosphorylated-Akt and an inhibition of cell proliferation via cell cycle arrest in the G0/G1 phase by P27/Kip1 accumulation, but there was no obvious induction of apoptosis. The simultaneous treatment of LY294002 and CH-11 significantly induced apoptosis confirmed by morphology and DNA ladder formation. Decreased phosphorylated-Akt by LY294002 treatment led to a down-regulation of Mcl-2 and phosphorylated Bad proteins, which are anti-apoptotic factors and belong to the Bcl-2 family. On the other hand, expression levels of the other anti-apoptotic factors, such as FLICE-inhibitory protein (FLIP), Bcl-2 and Bcl-XL, which are associated with the Fas-mediated apoptotic signal pathway, did not change after LY294002 treatment. We concluded that: 1) the PI3K-Akt pathway plays an important role in preventing Fas-mediated apoptosis; and 2) a PI3 K inhibitor, such as LY294002, might be a useful anti-tumoral agent for gastric carcinoma.

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Acknowledgements

We thank Dr. Hidetoshi Yamazaki (Division of Immunology, Tottori University) for his excellent technical advice regarding flow cytometry analysis. We also thank Mr. Norihisa Itaki, Ms Mayumi Kajimura, Ms Yoshiko Nishimiya, and Dr. Tonang Dwi Ardyanto, for their skillful technical assistance. This work was supported, in part, by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (grant number, 14370069).

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  1. Division of Organ Pathology, Department of Microbiology and Pathology, Faculty of Medicine, Tottori University, 86 Nishi-cho, Yonago, 683–8503, Tottori, Japan

    Mitsuhiko Osaki, Satoru Kase, Keiko Adachi, Ami Takeda, Kiyoshi Hashimoto & Hisao Ito

  2. Division of Molecular Genetics and Biofunction, Department of Biomedical Science, Graduate School of Medicine, Tottori University, Yonago, Japan

    Mitsuhiko Osaki

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  1. Mitsuhiko Osaki
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Correspondence to Mitsuhiko Osaki.

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Osaki, M., Kase, S., Adachi, K. et al. Inhibition of the PI3K-Akt signaling pathway enhances the sensitivity of Fas-mediated apoptosis in human gastric carcinoma cell line, MKN-45. J Cancer Res Clin Oncol 130, 8–14 (2004). https://doi.org/10.1007/s00432-003-0505-z

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