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Establishment of Paclitaxel-Resistant Cell Line and the Underlying Mechanism on Drug Resistance
  1. Jingjing Zhang, MD*,,
  2. Jin Zhao, MS*,
  3. Wenjing Zhang, MS,
  4. Guanyuan Liu, PhD§,
  5. Dongmei Yin, MS,
  6. Jian Li, MS*,
  7. Sumei Zhang, BS* and
  8. Hongxia Li, MD, PhD*
  1. *Department of Obstetrics and Gynecology, Beijing Shijitan Hospital, and
  2. Department of Gynecology, Beijing Obstetrics and GynecologyHospital, Capital Medical University;
  3. Department of Obstetrics and Gynecology, Beijing Military General Hospital; and
  4. §Department of Obstetrics and Gynecology, Beijing Chaoyang Hospital, Capital Medical University, Beijing, China.
  1. Address correspondence and reprint requests to Hongxia Li, MD, PhD, Department of Obstetrics and Gynecology, Beijing Shijitan Hospital of Capital Medical University, 10 Tieyi Road of Haidian District, Beijing 100038, China. E-mail: Lihx69@hotmail.com.

Abstract

Objective The objective of this study was to establish a taxol (TAX)–resistant human ovarian carcinoma cell line and investigate its drug-resistant mechanism.

Methods Using the dose calculated from clinical chemotherapy, we established a TAX-resistant human ovarian carcinoma cell line OC3/TAX300 by intermissive and repeated exposure to TAX of a high concentration at 300 μg/mL for 2 hours each time. The drug sensitivity was examined by tetrazolium dye (MTT) test. Distribution of cell cycle, DNA content analysis, and P-glycoprotein (P-gp) expression were detected by flow cytometry. We detected the differential gene expression by use of cDNA microarray. The reverse transcription–polymerase chain reaction and Western blot were used to verify the representative mRNA expression and their protein expression.

Results OC3/TAX300 cells were established after 10 months with stable resistance, and the drug resistance index was 6.70. It displayed significant cross-resistance to topotecan. Distribution of cell cycle revealed a higher percentage of G2 + M phase (P < 0.01), a lower percentage of S phase (P < 0.05), and overexpression of P-gp (P < 0.01). The cDNA microarray analysis showed that there were 134 significantly differential expression genes in all, of which up-regulated and down-regulated genes were 17 and 117, respectively. The up-regulated genes JAK2 (Janus kinase 2) and HSPC154 were confirmed by reverse transcription–polymerase chain reaction and Western blot.

Conclusions The OC3/TAX300 cell line is an ideal model to investigate the mechanism of TAX resistance. Taxol resistance in this cell could be related to overexpression of P-gp and the change of cell cycle profiles. The differential expression genes of JAK2 and HSPC154 may be candidate genes associated with TAX resistance in ovarian carcinoma cell lines.

  • Taxol resistance
  • Ovarian neoplasm
  • Gene expression

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Footnotes

  • This work was financially supported by the Capital Medical Development Fund of China (grant ZD199915).

  • The authors declare no conflict of interest.

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