Lewis肺癌耐药细胞的诱导及其耐药性的初步研究

Establishment and characteristics of multidrug-resistant Lewis lung cancer cell line

目的 建立肺癌耐药动物模型 ,诱导耐ADM的Lewis肺癌细胞株 ,并鉴定其生长特性、细胞内药物的释放特性及耐药谱。方法 用渐进式给药法诱导建立耐ADM的Lewis肺癌细胞株 ,绘制生长曲线 ,计算倍增时间 ;用IPP图象分析系统观察比较耐药细胞及其亲本细胞内ADM和Rh B的荧光光密度 ;荧光分光光度计法测定细胞内ADM含量并绘制ADM释放曲线 ;MTT法测定细胞的IC5 0 ,计算耐药指数。结果 经 14个月的诱导培养获得了能在 0 .4μg mlADM中稳定生长的Lewis肺癌细胞株———L3 8/ADM ,其在动物体内的成瘤率为 10 /10 ;在 0 .15mg/lADM和普通 164 0培养基中的倍增时间分别为 2 2 .0h和 2 1.8h ;在 4mg/lADM和Rh B中培养后 ,耐药细胞及其亲本细胞内的荧光光密度比分别为 2 .82∶1和 2 .65∶1,耐药细胞内的ADM含量仅为其亲本细胞的 64 .7% ,但其药物释放速度无明显差异。L3 8/ADM细胞除对ADM耐药外 ,对DNR、VCR、MIT和CDDP也有不同程度的耐药性。结论 L3 8/ADM是一株典型的MDR细胞株 ,其耐药作用可能是由于胞膜的药物渗透受阻所至。

Objective To establish an ADM resistant Lewis lung cancer cell line and to investigate its biological characteristics. Methods The multidrug resistant cell line was gradually induced by ADM from Lewis lung cancer cell line (L3 8), its growth characteristics and cancinogenicity were observed. The fluorescent density of ADM and Rh B in the cells were analyzed by IPP image analysis system. The ADM concentration in cells was assayed with fluorescent meter. The IC50 was evaluated by MTT assay and the drug resistant index was counted. Results The drug resistant Lewis lung cancer cell line, L3 8/ADM, which can grow in the medium containing 0.4?mg/l ADM was acquired after 14 months selective culture. Its tumor generatic rate was 10/10. When the L3 8/ADM was cultured in 0.15?mg/l ADM and general 1640 medium, the doubling time was 22.0?h and 21.8?h separately. After culturing in 4?mg/l ADM and Rh B medium, the fluorescent density ratio of L3 8/ADM and L3 8 were 2.82∶1 and 2.65∶1 separately. The ADM concentration of L3 8/ADM was only 64.7% of its parental, but there was no significant difference in their ADM release rate. Except for ADM, L3 8/ADM was resistant to DNR, VCR, MIT and CDDP in different degrees. Conclusion L3 8/ADM cell is a typical MDR cell line. The cell membrane obstruction of drug infiltration might be the cause of drug resistance. This study will provide a basis for the establishment of lung cancer MDR animal model.