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Establishment and characterization of monoclonal 5-fluorouracil-resistant cell lines derived from human endometrial
Tetsuji Tanaka1, Tao Bai, Saori Toujima
1Department of Obstetrics and Gynecology, Wakayama Medical University, 811-1 Kimi-idera, Wakayama 641-0012, Japan. obgywmu@wakayama-med.ac.jp
Abstract:
To investigate acquired 5-fluorouracil (5FU)-resistance in cancer cells, we established four monoclonal 5FU-resistant cell lines from human endometrial adenocarcinoma cells by long-term 5FU-exposure cultures and limiting dilution cultures. The established subclones exhibited 5-25 times greater 5FU-resistance than the parent cells, and showed suppression of 5FU-induced DNA fragmentation. All four 5FU-resistant subclones were 25-125 times more resistant to SN38, 4-hydroxy-cyclophosphamide, paclitaxel and etoposide than the parent cells while none of the four subclones showed resistance to mitomycin. Two of the four subclones showed no resistance to pirarubicin and bleomycin. Although all four 5FU-resistant subclones were 5-25 times more resistant to anti-Fas IgM than the parent cells, the resistance levels to anti-Fas IgM of the individual subclones did not coincide with the strengths of their multidrug resistance. Karyotyping analyses revealed that the parent cells and the three 5FU-resistant subclones examined had normal 46XX karyotypes. These results indicate that the cell death signals induced by mitomycin, pirarubicin and bleomycin are distinctly different from the Fas-mediated apoptotic signals, that acquisition of 5FU-resistance can occur without any large chromosomal deletions or rearrangements, and that there are several possible molecular changes during the acquisition of 5FU-resistance. The established cell lines represent useful tools for investigating the mechanisms and treatments of acquired 5FU-resistance.
Insights
Researchers developed 5-fluorouracil (5FU)-resistant cancer cell lines to study drug resistance. These cell lines show cross-resistance to various chemotherapy drugs, aiding research into acquired 5FU-resistance mechanisms.
Area of Science:
- Oncology
- Cancer Cell Biology
- Pharmacology
Background:
- Acquired resistance to chemotherapy, such as 5-fluorouracil (5FU), is a major challenge in cancer treatment.
- Understanding the mechanisms of acquired drug resistance is crucial for developing effective therapeutic strategies.
Purpose of the Study:
- To establish and characterize 5-fluorouracil (5FU)-resistant human endometrial adenocarcinoma cell lines.
- To investigate the cross-resistance patterns of these 5FU-resistant cell lines to other chemotherapeutic agents.
- To explore the potential chromosomal and molecular changes associated with the acquisition of 5FU-resistance.
Main Methods:
- Generation of 5FU-resistant cell lines through long-term 5FU exposure and limiting dilution cultures.
- Assessment of 5FU-resistance levels and 5FU-induced DNA fragmentation.
- Evaluation of cross-resistance to SN38, 4-hydroxy-cyclophosphamide, paclitaxel, etoposide, mitomycin, pirarubicin, bleomycin, and anti-Fas IgM.
- Karyotyping analysis of parent and resistant cell lines.
Main Results:
- Four monoclonal 5FU-resistant sublines were established, exhibiting 5-25 fold greater 5FU-resistance and suppressed 5FU-induced DNA fragmentation.
- Significant cross-resistance (25-125 fold) to SN38, 4-hydroxy-cyclophosphamide, paclitaxel, and etoposide was observed.
- No resistance to mitomycin was found; variable resistance to pirarubicin and bleomycin was noted.
- Resistance to anti-Fas IgM (5-25 fold) did not correlate with multidrug resistance levels.
- Karyotyping revealed normal 46XX karyotypes in parent and most resistant cells, suggesting resistance acquisition without major chromosomal alterations.
Conclusions:
- Acquired 5FU-resistance can occur without significant chromosomal aberrations.
- Distinct cell death pathways exist for mitomycin, pirarubicin, bleomycin, and Fas-mediated apoptosis.
- The established 5FU-resistant cell lines are valuable tools for studying acquired drug resistance mechanisms and developing new treatments.
