J Korean Ophthalmol Soc.  2011 Feb;52(2):233-240.

Effect of Mitomycin C and 5-Fluorouracil on Cultured Human Nasal Mucosa Fibroblasts

• Kim KH
• Kim IT

Abstract

PURPOSE
To investigate the effects of two antimetabolites, mitomycin C (MMC) and 5-fluorouracil (5-FU), on proliferation of cultured human nasal mucosa fibroblasts.
METHODS
Human nasal mucosa fibroblasts were primarily cultured, and exposed to various concentrations of MMC and 5-FU for 5 minutes. Control fibroblasts were exposed to only DMEM media without the drugs. Effect of drugs on cell morphology was observed by phase-contrast microscopy. Cell viability and apoptosis were measured using MTT [3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyl-tetrazolium bromide] assay and Acridine orange/Hoechst (AO/HO) staining, respectively.
RESULTS
In both experimental groups exposed to MMC and 5-FU, fibroblasts maintained standard spindle shape. The MTT assay showed that both MMC and 5-FU inhibited fibroblast proliferation in a dose dependent manner. AO/HO staining showed apoptotic cells in both experimental groups.
CONCLUSIONS
Both MMC and 5-FU have an antiproliferative effect on fibroblasts in vitro at least through induction of apoptosis. Therefore, adjuvant use of either MMC or 5-FU during endonasal dacryocystorhinostomy may improve the clinical outcome by inhibiting proliferation of the nasal mucosa.

Keyword

5-fluorouracil; Mitomycin C; Nasal mucosa fibroblast

MeSH Terms

Antimetabolites
Apoptosis
Cell Survival
Dacryocystorhinostomy
Fibroblasts
Fluorouracil
Humans
Microscopy, Phase-Contrast
Mitomycin
Nasal Mucosa
Antimetabolites
Fluorouracil
Mitomycin

Figure

• 

  Figure 1. The morphology of cultured human nasal mucosa fibroblasts under phase-contrast microscope. Fibroblasts were treated with various concentrations of MMC for 5 minutes and then incubated. Images are representatives of day 1 of culture. The viable cells were spindle-shaped at all concentrations of MMC. Magnification, ×100. MMC = mitomycin C.

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  Figure 2. The morphology of cultured human nasal mucosa fibroblasts under phase-contrast microscope. Fibroblasts were treated with various concentrations of 5-FU for 5 minutes and then incubated. Images are representatives of day 1 of culture. The viable cells were spindle-shaped at all concentrations of 5-FU. Magnification, ×100. 5-FU=5-fluorouracil.

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  Figure 3. The viability of the cultured human nasal mucosa fibroblasts treated with various concentrations of MMC. Fibroblasts were treated for 5 minutes with MMC (0.01, 0.1, 0.2, 0.4 mg/ml), washed and further incubated for 24 hr. Optical density (OD) of the fibroblasts was measured using MTT assay. Growth rates (%) were calculated as [100 × (OD values of drug-treated cells/OD values of non-treated cells)]. Values and bars represent means of growth rates and the SD, respectively. ∗ p<0.05 compared to non-treated control. MMC = mitomycin C.

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  Figure 4. The viability of the cultured human nasal mucosa fibroblasts treated with various concentrations of 5-FU. Fibroblasts were treated for 5 minutes with 5-FU (0.5, 5, 10, 25 mg/ml), washed and further incubated for 24 hr. Optical density (OD) of the fibroblasts was measured using MTT assay. Growth rates (%) were calculated as [100 × (OD values of drug-treat-ed cells/OD values of non-treated cells)]. Values and bars represent means of growth rates and the SD, respectively. ∗ p<0.05 compared to non-treated control. 5-FU = 5-fluorouracil.

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  Figure 5. Acridine orange/Hoechst (AO/HO) staining of human nasal mucosa fibroblasts after treatment with MMC and 5-FU. Fibroblasts were not-treated (A), or treated with 0.4 mg/ml of MMC (B) and 25 mg/ml of 5-FU (C) for 5 minutes, washed and further incubated for 3 hr. The cells were then stained with AO/HO to identify apoptosis as shown in the Methods. (A) Normal control cells. Arrow indicates blue nuclei and brilliant red cytoplasm. (B) MMC-treated cells. The apoptotic cell shows orange-red nuclei, becoming indistinguishable from the cytoplasm. (C) 5-FU-treated cells. The apoptotic cell shows orange-red nuclei, becoming indistinguishable from the cytoplasm. Magnification, ×400. MMC = mitomycin C, 5-FU = 5-fluorouracil.

Reference

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