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Reversal effect of ouabain on multidrug resistance in esophageal carcinoma EC109/CDDP cells by inhibiting the translocation of Wnt/β-catenin into the nucleus

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Tumor Biology

Abstract

The incidence of esophageal carcinoma is increasing throughout the world. A major obstacle to its treatment is acquired multidrug resistance (MDR) which results in the failure of chemotherapy and patient relapse. Here, we identified that ouabain is capable of reversing MDR to cisplatin (CDDP) in EC109/CDDP cells and explore the possible mechanisms of action. The parental and the MDR cell lines were both sensitive to ouabain with 50 % inhibitory concentration (IC50) values of 258.11 and 710.63 nM, respectively. Cisplatin cytotoxicity increased in the EC109/CDDP cells by the addition of ouabain which helps promote CDDP-induced apoptosis. Ouabain at 20 nM effectively reduced the IC50 of CDDP in EC109/CDDP cells from 36.54 to 3.39 μM. This represents a 10.78-fold increase in sensitization to CDDP. We also found that ouabain was capable of down regulating the expression of P-glycoprotein (P-gp) and Bcl-2 in a dose- and time-dependent manner. Finally, the results indicated that ouabain suppressed Wnt luciferase report (TOPFlash) activity obviously in EC109/CDDP and depressed the translocation of β-catenin into the nucleus. Moreover, ouabain showed reversal effect of MDR to CDDP in nude mouse xenograft model, and reduced the protein level of β-catenin (Y333) in tumor tissue of CDDP plus ouabain group. All data proved that ouabain has a potent β-catenin-dependent anti-MDR effect.

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Acknowledgments

The study was supported by Jiangsu Provincial Special Program of Clinical Medical Science (BL2014040), Suzhou Science and Technology Development Program (SZS201509), and Natural Science Foundation of the Jiangsu Higher Education Institutions of China (11KJB180010).

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Authors and Affiliations

  1. Department of Radiotherapy and Oncology, The Second Affiliated Hospital of Soochow University, San Xiang Road No. 1055, Suzhou, Jiangsu, 215004, China

    Yucheng Shen & Ye Tian

  2. Institute of Radiotherapy and Oncology, Soochow University, San Xiang Road No. 1055, Suzhou, Jiangsu, 215004, China

    Yucheng Shen & Ye Tian

  3. Suzhou Key Laboratory for Radiation Oncology, San Xiang Road No. 1055, Suzhou, Jiangsu, 215004, China

    Yucheng Shen & Ye Tian

  4. Department of Oncology, Affiliated Haian Hospital of Nantong University, Zhongba Middle Road, No. 17, Haian Town, Haian County, Nantong, Jiangsu, 226601, China

    Yucheng Shen

  5. Laboratory Animal Center of Nantong University, Qi Xiu Road No.19, Nantong, Jiangsu, 210061, China

    Qinghua Wang

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  1. Yucheng Shen
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Correspondence to Ye Tian.

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All animal studies were carried out in compliance with the Guide for the Care and Use of Laboratory Animals of Nantong University, China.

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Yucheng Shen and Qinghua Wang have contributed equally to this study.

Electronic supplementary material

Fig. 1

Effect of ouabain on EC109/CDDP cell cycle was detected by flow cytometry and western blot. (GIF 94 kb)

High resolution image (TIFF 309 kb)

Fig. 2

H&E stains the structure of mice liver. (GIF 303 kb)

High resolution image (TIFF 1020 kb)

Table. 1

Examine the level of ALT and AST in mice liver by ELISA. (GIF 14 kb)

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Shen, Y., Wang, Q. & Tian, Y. Reversal effect of ouabain on multidrug resistance in esophageal carcinoma EC109/CDDP cells by inhibiting the translocation of Wnt/β-catenin into the nucleus. Tumor Biol. 37, 15937–15947 (2016). https://doi.org/10.1007/s13277-016-5437-8

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  • DOI: https://doi.org/10.1007/s13277-016-5437-8

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