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目的观察接种密度和α-二氟甲基鸟氨酸(DFMO)对小肠上皮细胞(IEC-6)体外增殖的影响.方法 IEC-6 细胞以不同密度接种于96孔培养板,加入或不加DFMO.细胞接种后第1, 2, 3, 4, 5 , 6和7天,分别用细胞计数板直接计数细胞数量,用噻唑蓝比色法(MTT法)测定波长570? nm处的光密度(OD)值,确定细胞的增殖活力.结果 OD值和细胞数量具有正相关(r=0.954,P<0.01);较高接种密度(>0.5×104 细胞/孔)组接种后第2天,细胞生长抑制,尤其是密度增加到4×104 细胞/孔时,第2～第 3天OD值逐渐增加,第4天明显增加,第5天达高峰,此后OD值开始下降.而低密度(0.2× 104细胞/孔)接种后第4天,细胞生长出现抑制,此后其生长与其它密度相似.另外,加入 DFMO后1～3天,完全抑制了细胞增殖,此后与对照组一样,细胞逐渐生长.结论 IEC-6 细胞增殖与接种密度和培养时间有关;随着培养时间的延长,DFMO抑制细胞增殖作用具有可逆性,其与培养时间延长有关.

Objective To characterize the role of plating densities and alpha-difluoromethylornithine (DFMO) on the proliferation of IEC-6 cells in vitro.Methods IEC-6 cells were seeded in 96-well microplates at various densities in the presence or absence of DFMO. Cells were counted and their proliferative capability was monitored Days 1 to 7 with MTT assay at an optical density of 570nm.Results There was a positive relationship between cell number and OD value (r=0.954, P<0.01). Higher plating densities (>0.5×104 cells/well) inhibited the growth of cells on Day 2. When the density reaches 4×104 cells/well, the OD value increased gradually and reached a peak on Day 5. After that, the OD value began to fall. The growth of IEC-6 cells was limited at a low density (0.2×104 cells/well) on Day 4. DFMO caused a complete inhibition of proliferation of IEC-6 cells on Days 1 to 3. Conclusion Proliferation of IEC-6 cells is related to plating density and incubation time. It is inhibited by DFMO, but is reversible when the incubation time is prolonged.