[Establishment of methotrexate-resistant human choriocarcinoma cell line and its biological characteristics].
To establish a methotrexate (MTX)-resistant choriocarcinoma cell line and to determine its biologic characteristics.



MTX-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.



JAR/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.



JAR/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.
Chen
Ya-xia
YX
The Affiliated Obstetrics and Gynecology Hospital, College of Medicine  Zhejiang University, Hangzhou 310006, China.
Xia
Xing
X
Chen
Huai-zeng
HZ
Cheng
Qi
Q
Zhou
Cai-yun
CY
D017304:Annexin A5 / Q000032:analysis; D017209:Apoptosis; D002448:Cell Adhesion; D002455:Cell Division; D045744:Cell Line, Tumor; D002822:Choriocarcinoma / Q000188:drug therapy / Q000235:genetics / Q000473:pathology*; D019008:Drug Resistance, Neoplasm; D006801:Humans; D008727:Methotrexate / Q000494:pharmacology*
D017304:Annexin A5; D008727:Methotrexate