Establishment of methotrexate-resistant human choriocarcinoma cell line and its biological characteristics.
- Author:
Ya-xia CHEN
1
;
Xing XIA
;
Huai-zeng CHEN
;
Qi CHENG
;
Cai-yun ZHOU
Author Information
- Publication Type:Journal Article
- MeSH: Annexin A5; analysis; Apoptosis; Cell Adhesion; Cell Division; Cell Line, Tumor; Choriocarcinoma; drug therapy; genetics; pathology; Drug Resistance, Neoplasm; Humans; Methotrexate; pharmacology
- From: Journal of Zhejiang University. Medical sciences 2004;33(2):138-165
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish a methotrexate (MTX)-resistant choriocarcinoma cell line and to determine its biologic characteristics.
METHODSMTX-resistant cell line (JAR/MTX) was derived from human choriocarcinoma cell line JAR by exposed to intermittently and progressively increasing concentration of MTX. Drug sensitivity was detected by MTT; P-gp GST-Pi and PCNA expressions were detected by immunohistochemistry. Cell apoptosis was detected by flow cytometry (FCM) with PI/Annexin V stain. Growth rates and human chorionic gonadotropin (HCG) production were also measured.
RESULTSJAR/MTX cell line was established with stable MTX-resistance (resistance index to MTX was 7.3) and cross-resistant to TAX and VCR. Growthrate of JAR/MTX was lower than that of parent cell line JAR. Expression level of PCNA in JAR/MTX was lower than that in JAR (3.09+/-0.42 compared with 3.72+/-0.35, P<0.05), while GST-pi expression was higher. No statistical difference of P-gp expression existed between two cell lines. JAR/MTX secreted more HCG than JAR every 10(5) cells secreted (95.7+/-5.4 compared with 41.3+/-2.8)mIU after 48 h(P<0.01). The flow cytometry showed that the spontaneous and MTX induced apoptosis in JAR/MTX was significantly lower than that in JAR P<0.05.
CONCLUSIONJAR/MTX cell line presented stable resistant to MTX and cross-resistant to TAX and VCR, which might sever as a model in study of drug resistance in choriocarcinoma.
