Establishment of paclitaxel-resistant cell line and the underlying mechanism on drug resistance.
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.
Department of Obstetrics and Gynecology, Beijing Shijitan Hospital of Capital Medical University, 10 Tieyi Road, Haidian District, Beijing, China.
Jingjing
J
Zhang
Jin
J
Zhao
Wenjing
W
Zhang
Guanyuan
G
Liu
Dongmei
D
Yin
Jian
J
Li
Sumei
S
Zhang
Hongxia
H
Li
D000970:Antineoplastic Agents / Q000627:therapeutic use; D000077216:Carcinoma, Ovarian Epithelial; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D005260:Female; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D046228:Microarray Analysis; D009375:Neoplasms, Glandular and Epithelial / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology*; D010051:Ovarian Neoplasms / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology*; D017239:Paclitaxel / Q000627:therapeutic use*; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics
D000970:Antineoplastic Agents; D017239:Paclitaxel