Effect of the cyclin-dependent kinases inhibitor p27 on resistance of ovarian cancer multicellular spheroids to anticancer chemotherapy (original) (raw)

Abstract

Purpose: A low proliferating fraction in solid tumors limits the effectiveness of cell-cycle-dependent chemotherapeutic agents. To understand the molecular basis of such resistance, we examined the expression of the cyclin-dependent kinases inhibitor p27, and relationship with drug resistance and P-gp expression in ovarian cancer multicellular spheroids. Methods: We cultured ovarian cancer cells (A2780 and CAOV3) as multicellular spheroids and examined the expression of p27 and P-glycoprotein (P-gp) by western blot, flow cytometry and confocal. We also analyzed the cell-cycle distribution by flow cytometry. In addition, trypan blue exclusion testing and cell apoptosis analysis were used to detect the sensitivity to Taxol. Results: When transferred from monolayer to three-dimensional culture, a consistent upregulation of p27 protein and P-gp protein was observed in ovarian cancer cell lines. Compared with monolayer cells, there was a significant increase of G0-G1 phase cells and decrease of S and G2-M phase cells in spheroid cells. Aggregates of cells showed higher cell viability than monolayer cells. Antisense oligodeoxynucleotide (ASON) -mediated downregulation of p27 reduced intercellular adhesion, increased cell proliferation, downregulated P-gp expression and sensitized cells to Taxol. Conclusions: Our results implicate that p27 serves as a regulator of drug resistance in ovarian tumors. ASON-mediated alteration of p27 reverses resistance of ovarian cancer to anticancer agents that are associated with increased sensitivity of ovarian cancer cells to chemotherapeutic agents.

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Abbreviations

MCS:

Multicellular spheroids

ASON:

Antisense oligodeoxynucleotide

SON:

Sense oligodeoxynucleotide

SON/monolayer:

Monolayer cells treated with SON

ASON/monolayer:

Monolayer cells treated with ASON

SON/MCS:

Spheroid cells treated with SON

ASON/MCS:

Spheroid cells treated with ASON

P-gp:

P-glycoprotein

MDR:

Multidrug resistance

BSA:

Bovine serum albumin

PI:

Propidium iodide

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Acknowledgements

This work was supported by National Science Foundation of China (No. 39870765) and ‘973’ program of China (No. 2002CB513100)

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Authors and Affiliations

1. Department of Obsterics and Gynecology, Tongji Hospital Affiliated to Tongji Medical School, Huazhong University of Science and Technology, Wuhan, Hubei, 430030, People Republic of China
   Hui Xing, Shixuan Wang, Jing Li, Qinglai Gao, Xiaokui Yang, Danhui Weng, Yunpin Lu & Ding Ma
2. Division of Gastroenterology/Hepatology, University of California, Irvine Medical College, Orange, CA, 92868, People Republic of China
   Keqin Hu
3. Department of Orthepeadecs, Xiangfan Hospital Affiliated to Tongji Medical School, Huazhong University of Science and Technology, Xiangfan, Hubei, 441021, People Republic of China
   Wenming Tao

Authors

1. Hui Xing
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2. Shixuan Wang
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3. Keqin Hu
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4. Wenming Tao
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5. Jing Li
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6. Qinglai Gao
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7. Xiaokui Yang
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8. Danhui Weng
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9. Yunpin Lu
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10. Ding Ma
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Correspondence toDing Ma.

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Hui Xing and Shixuan Wang have contributed equally in this work

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Xing, H., Wang, S., Hu, K. et al. Effect of the cyclin-dependent kinases inhibitor p27 on resistance of ovarian cancer multicellular spheroids to anticancer chemotherapy.J Cancer Res Clin Oncol 131, 511–519 (2005). https://doi.org/10.1007/s00432-005-0677-9

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• Received: 30 August 2004
• Accepted: 14 February 2005
• Published: 28 May 2005
• Issue Date: August 2005
• DOI: https://doi.org/10.1007/s00432-005-0677-9

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