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[Establishment of methotrexate-resistant human choriocarcinoma cell line and its biological characteristics]
Ya-xia Chen1, Xing Xia, Huai-zeng Chen
1The Affiliated Obstetrics and Gynecology Hospital, College of Medicine Zhejiang University, Hangzhou 310006, China.
Objective:
To establish a methotrexate (MTX)-resistant choriocarcinoma cell line and to determine its biologic characteristics.
Methods:
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.
Results:
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.
Conclusion:
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.
Insights
A new methotrexate-resistant choriocarcinoma cell line (JAR/MTX) was developed. This model exhibits cross-resistance to other drugs and altered biological characteristics, aiding choriocarcinoma drug resistance studies.
Area of Science:
- Oncology
- Pharmacology
- Cell Biology
Context:
- Choriocarcinoma is a rare malignancy with varying responses to chemotherapy.
- Methotrexate (MTX) is a key chemotherapeutic agent, but resistance can limit its efficacy.
- Developing drug-resistant cell lines is crucial for understanding resistance mechanisms.
Purpose:
- To establish a methotrexate (MTX)-resistant choriocarcinoma cell line (JAR/MTX).
- To characterize the biologic properties of the established resistant cell line.
- To evaluate cross-resistance patterns and changes in cellular markers.
Summary:
- A JAR/MTX cell line was successfully derived from the parental JAR choriocarcinoma line.
- The JAR/MTX line demonstrated stable MTX resistance (7.3-fold) and cross-resistance to taxane (TAX) and vincristine (VCR).
- Compared to JAR cells, JAR/MTX cells showed reduced proliferation, lower PCNA expression, higher GST-pi expression, and significantly lower apoptosis rates, alongside increased human chorionic gonadotropin (HCG) secretion.
Impact:
- The JAR/MTX cell line serves as a valuable in vitro model for investigating drug resistance in choriocarcinoma.
- Understanding the biologic characteristics of this resistant line can inform therapeutic strategies.
- This model facilitates research into the molecular mechanisms underlying chemoresistance in gestational trophoblastic diseases.
