阿霉素对人胆管癌细胞株FRH-0201的生物学特性的影响

The effects of adramycin on the human liver hilar cholangiocarcinona cell line FRH-0201

目的观察阿霉素长时间作用对胆管癌细胞株FRH-0201生物学活性的影响。方法用2μg/ml的阿霉素同一浓度逐渐延长时间作用于人胆管癌细胞株FRH-0201细胞,经反复作用筛选出能在含1μg/ml的阿霉素培养液中生存72h,去掉阿霉素后仍能继续稳定传代生长的细胞。无药培养1月后进行生物学行为的监测。观察细胞形态学,绘制生长曲线和流式细胞仪测定细胞周期,酶联免疫吸附法测定细胞的肿瘤标记物,激光共聚焦显微镜观察细胞内的阿霉素的荧光强度。结果阿霉素作用后FRH-0201细胞形态学有轻度改变,细胞倍增时间较亲本细胞延长3h,与亲本细胞相比S期细胞增多16.89%,G1期细胞减少29.33%、G2期细胞增多12.46%。CA125和CA19-9试验组和亲本组分别为9.14和8.60U/ml,1.59和2.96U/ml,细胞内阿霉素浓度(荧光强度平均值),亲本细胞为1764.8,试验组细胞为305.4。结论阿霉素对胆管癌细胞株FRH-0201的形态学及倍增时间无明显影响,但使细胞进入S期、G2期的增多,细胞内药物浓度明显降低,生长时间无明显变化。

Objective ： To investigate the effects of adramycin on ceil line of FRH-0201. Methods ： Human hilar oholangiooaroinona cell line were cultured with adramyoin （2μg/ml） through lasting different times,finally this cell line can survive after 72h cultured in the culture liquid with the adramyoln（1μg/ml）. Then the medium was replaced with a fresh one without adramycin,ancl the cells were continuously cultured for 1 month. The in vitro studies included the observation of the appearance with optical microscope, MTT cell proliferation assay, analysis of the Cell Cycle by Flow cytometry（FAOS）, assays of tumor marker using enzyme-linked immunosor bent assay（ELISA）, the fluorescence density of adramycin. Results ： Compared to the parent FRH-0201 cell line,The morphology showed typical morphological characteristics,the doubling time prolonged about, 3h, the cell cycle by flow oytometry identified in G1,G2 and S phase of cell cycle were 40.50%, 19.74% and 39.77% in trial group, and 69.83% ,7.29% and 22.88% in parent group, respectively , Tumor marker has no difference batween them, the fluorescence density of epirubicin desoending 5.2 times. Concusion： The adramycin can affect the cell cycle and cause the change of metabollsm.