Establishment of methotrexate-resistant human choriocarcinoma cell line and its biological characteristics / 浙江大学学报·医学版

ABSTRACT

<p><b>OBJECTIVE</b>To establish a methotrexate (MTX)-resistant choriocarcinoma cell line and to determine its biologic characteristics.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>