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Administration sequence-dependent antitumor effects of paclitaxel and 5-fluorouracil in the human gastric cancer cell line MKN45

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Abstract

Background: The clinical outcome of gastric cancer patients has been improved by combination of 5-fluorouracil (5-FU) and paclitaxel (PXL). However, the optimal schedule of this combination has not been determined. Methods: The efficacies of sequential administrations of 5-FU and PXL on the gastric cancer cell line MKN45 were investigated using a WST-8 colorimetric assay. The cell cycle distribution of each drug was evaluated by flow-cytometry. Furthermore, the mechanism of antitumor activity enhancement by the administration sequence was investigated by western blotting. Results: MKN45 cell growth was significantly inhibited by each drug in a dose- and time-dependent manner. The cytotoxicities of PXL followed by 5-FU were significantly greater than those of 5-FU followed by PXL. The flow-cytometric analysis revealed that PXL exposure caused viable cell accumulation in G2/M phase in a dose-dependent manner. Western blotting showed that PXL exposure followed by 5-FU up-regulated Chk1 and Wee1 protein expressions until PXL removal and 5-FU exposure, when these expressions gradually decreased to their basal levels. 14-3-3σ protein expression was significantly up-regulated upon PXL treatment followed by 5-FU. Interestingly, Mad2 protein expression with PXL treatment followed by 5-FU gradually increased after the PXL removal and 5-FU exposure. Conclusions: PXL followed by 5-FU administration may be the optimal sequence for treatment of gastric cancer. The enhanced viable cell accumulation after PXL pretreatment may be related to G2 arrest. After PXL removal and 5-FU exposure, the cells progressing to M phase may undergo cell death by mitotic catastrophe due to DNA damage caused by 5-FU exposure.

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Authors and Affiliations

  1. The Second Department of Surgery, Mie University School of Medicine, 2-174 Edobashi, Tsu, Mie, 514-8507, Japan

    Yuji Toiyama, Koji Tanaka, Naomi Konishi, Yasuhiko Mohri, Chikao Miki & Masato Kusunoki

  2. Department of Innovative Surgery, Mie University School of Medicine, 2-174 Edobashi, Tsu, Mie, 514-8507, Japan

    Hitoshi Tonouchi & Masato Kusunoki

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  1. Yuji Toiyama
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  2. Koji Tanaka
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  3. Naomi Konishi
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  4. Yasuhiko Mohri
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  5. Hitoshi Tonouchi
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  6. Chikao Miki
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  7. Masato Kusunoki
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Correspondence to Masato Kusunoki.

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Toiyama, Y., Tanaka, K., Konishi, N. et al. Administration sequence-dependent antitumor effects of paclitaxel and 5-fluorouracil in the human gastric cancer cell line MKN45. Cancer Chemother Pharmacol 57, 368–375 (2006). https://doi.org/10.1007/s00280-005-0057-9

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  • DOI: https://doi.org/10.1007/s00280-005-0057-9

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