人肺癌细胞5-氟脲嘧啶耐药株的建立及其生物学特性分析

Development and Characterization of 5-fluorouracil Resistant Human Lung Cancer Cell Line H460/5-FU

建立人肺癌5-氟脲嘧啶(5-Fluorouracil,5-FU)的耐药细胞株H460/5-FU,并对其生物学特性进行分析。实验采用浓度梯度递增法诱导肺癌细胞耐药,建立肺癌细胞耐药模型。采用四甲基偶氮唑蓝法(MTT),计算H460和H460/5-FU的半数抑制浓度(IC50)及耐药系数(RI);通过流式细胞仪分析H460和H460/5-FU的细胞周期分布;光镜观察细胞形态变化;通过流式细胞仪进行SP(side population)细胞检测,比较SP细胞所占比例;做细胞生长曲线和集落形成实验观察细胞生长及倍增情况。结果表明5-FU对H460和H460/5-FU的IC50分别为28.7μg/mL和90.44μg/mL,RI为3.15;H460/5-FU细胞倍增时间缓慢,DNA复制时间延长,且含有更多的SP细胞。

The paper establishes a 5-FU-resistant human lung cancer cell line and analyses its characterization. The H460/5-FU cell line is obtained by increasing the concentration of 5-fluorouracil gradually; MTT assay is used to measure and compare IC50 and RI between H460/5-FU cell line and its parental cell line---H460;Cetl cycle distribution of both cell lines is examined by flow cytometry;Morphological changes between the two cell lines are observed by the microscope;Then, flow cytometry is used again to detect the proportion of side population cells in the two cell lines;The cell growth curve and colony forming experiments are made to learn their proliferation. The IC50 and RI of H460/5-FU cell line are 90.44 μg/mL and 3.15, and it spends more time on proliferation and DNA replication than its parental cell line; Besides, the number of side population cells in H460/5-FU is more than that of H460 cell line.