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Establishment of a 5-fluorouracil-resistance laryngeal cancer cell line and its biology characteristics
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Abstract:
Objective:To establish a multidrug resistance cell line from human laryngeal cancer cells by 5-fluorouracil (5-Fu), and provide a research model of drug resistance mechanisms of laryngeal cancer. Methods: A multidrug resistance cell line from human laryngeal Hep-2 cancer cells (Hep-2/5-FU) was established by a high dosage intermittent increase gradient method. The morphology and cell doubling time of Hep-2 and Hep-2/5-FU cells was compared; the IC50 and drug resistance fold was detected by MTT assay; the cell cycle distribution and rhodamine accumulation in Hep-2 and Hep-2/5-Fu cells was studied by flow cytometry; and the MDR1 mRNA and the corresponding MDR1-P protein expressions were detected by real-time quantitative RT-PCR or Western blotting.Results: Hep-2/5-Fu cells was established，which had a stable resistance to 5-Fu; Hep-2/5-Fu cells exhibited cross resistance to chemotherapeutic agents like cisplatin and vincristine, and its doubling time was prolonged as compared with Hep-2 cells (\[31.25±4.37\] h vs \[25.62±353\] h, P<0.05). The proportion of cells in G0/G1 phase was increased (\[45.6±3.4\]% vs \[30.5±1.2\]%, P<0.05), while decreased in S phase (\[32.1±4.2\]% vs \[52.4±3.6\]%, P<0.05) in Hep-2/5-Fu cells as compared with those in Hep-2 cells; rhodamine accumulation in Hep-2 cells was much higher than that in Hep-2/5-Fu cells(\[89.83±0.52\]% vs \[14.38±0.48\]%, P<0.01); and the expressions of MDR1 mRNA (\[13.69±1.12\] vs \[1782±061\], P<0.05) and the corresponding MDR1-P protein were increased in Hep-2/5-Fu cells than those in Hep-2 cells. Conclusion: The Hep-2/5-Fu cell line shows the typical and stable multidrug resistance, and may serve as a research model of laryngeal cancer resistance mechanism.
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Project Supported:
Project supported by the Science Research Development Fundation of Shanghai Renji Hospital (No. RJPY10-011)
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