Effect of brief exposure to mitomycin C on cultured human nasal mucosa fibroblasts
Effect of brief exposure to mitomycin C on cultured human nasal mucosa fibroblasts
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DOI：
10.1097/00002341-200003000-00006
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发表时间：
2000-03-01
影响因子：
2
通讯作者：
Oda, D
中科院分区：
文献类型：
--
作者：
Hu, D;Sires, BS;Oda, D
目的：观察丝裂霉素C(MMC)对培养的人鼻粘膜成纤维细胞的影响。方法：将培养的人鼻粘膜成纤维细胞暴露于MMC(0.1~0.4 mg/ml)1~5min。用四甲基偶氮唑蓝(3-[4，5-二甲基噻唑-2-基]-2，5-二苯基四氮唑溴化法)检测细胞活力，末端脱氧核苷转移酶介导的dUTP缺口末端标记法(TUNEL)检测DNA片段化(细胞凋亡)，流式细胞仪检测细胞凋亡率，光镜下观察细胞形态。结果：部分细胞在丝裂霉素作用下存活，并在2~3d内再生。这些细胞在5~7天内达到融合。用四甲基偶氮唑盐比色法测定0.4 mg/mlMMC作用5min的抑制率为31.3%。浓度越低，作用时间越短，抑制率越低(但不显著)，呈现剂量-反应效应。与正常对照组相比，MMC(0.4 mg/ml)作用5min后24小时可见成纤维细胞DNA片段化。0.4 mg/mlMMC处理组的细胞凋亡率明显高于对照组(p<；0.05)。结论：短时间的MMC作用对鼻黏膜成纤维细胞有不同程度的细胞毒作用，并抑制其增殖。MMC还可以诱导细胞凋亡，作用时间为5分钟。因此，丝裂霉素C在泪囊鼻腔吻合术中可能具有复杂的作用。
Purpose: To observe the effect of mitomycin C (MMC) on cultured human nasal mucosa fibroblasts.Methods: Cultured human nasal mucosa fibroblasts were exposed to MMC (0.1-0.4 mg/ml) for 1 to 5 minutes. The viability of the fibroblasts was determined by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay; DNA fragmentation (apoptosis) by terminal deoxynucleotldyl transferase-mediated dUTP nick-end labeling (TUNEL); apoptotic percentage by flow cytometry; and morphology by light microscopy.Results: A portion of the fibroblasts survived the mitomycin treatment and showed evidence of regrowth within 2 to 3 days. These cells reached confluence in 5 to 7 days. The inhibition rates by MTT assay of 0.4 mg/ml MMC for 5-minute exposures was 31.3%. Dose-response effect was noted with the lower concentrations and shorter exposure times where the inhibition rates were lower (but not significantly so). DNA fragmentation was observed in fibroblasts 24 hours after MMC exposure (0.4 mg/ml) for 5 minutes compared with normal control. The apoptotic rate of fibroblasts treated by 0.4 mg/ml MMC was significantly higher than the control (p < 0.05).Conclusions: Short MMC exposure times have a variable cytotoxic effect and inhibit proliferation of cultured nasal mucosa fibroblasts. MMC also can induce apoptosis with a 5-minute exposure time. Therefore, it is possible that MMC has a complex effect in dacryocystorhinostomy.