抗阿霉素人食管癌Eca-109细胞生物学特性研究

Study on the biological features of human esophageal cancer cell resistant to adriamycin

目的： 建立并研究抗阿霉素人食管癌细胞株（Ｅｃａ１０９／Ａｄｒ） 的生物学特性。方法： 采用递加培养液中药物浓度建立Ｅｃａ１０９／Ａｄｒ 细胞株，测试该细胞株对多种抗肿瘤药物的敏感性，采用光镜及扫描电镜观察其形态变化，采用聚合酶链反应（ＰＣＲ） 分析法检测ｍｄｒ１ 表达，采用荧光分光光度法检测细胞内ＧＳＨ 水平。结果： 建立了低程度的抗阿霉素Ｅｃａ１０９／Ａｄｒ 细胞株，该细胞株细胞大小不均匀，边界不如Ｅｃａ１０９ 细胞整齐，有明显皱褶，未见微毛，集落形成率明显下降，对长春新碱、放线菌素Ｄ、三尖杉酯碱、丝裂霉素Ｃ、鬼臼乙叉甙及顺铂呈不同抗性，对氟尿嘧啶仍敏感，ｍｄｒ１ 基因呈低表达，细胞内ＧＳＨ 水平升高，结论：Ｅｃａ１０９／Ａｄｒ 细胞为多药抗药性细胞株。

Objective: To establish the adriamycin resistant human esophageal cancer cell line (Eca 109/Adr) and to study its biological features. Methods: Eca 109/Adr cell line was established by gradually increasing the adriamycin (Adr) concentration in the culture medium and the drug sensitivity of this cell line to other anticancer drugs was tested. The morphology of Eca 109/Adr cells was observed under scanning electron microscope. The multidrug resistance gene (mdr 1 gene) was detected by polymerase chain reaction (PCR) analysis and the level of intracellular glutathione (GSH) was measured by fluorescence spectophtometer.Results: The resistant Eca 109 cells to adriamycin with low grade were screened by treating the sensitive cells with adriamycin for a long time in the absence of mutagenic agents. The resistant cells had cross resistance to VCR, Act D, MMC, VP 16 and DDP but they were still sensitive to 5 FU. Compared with Eca 109 cells, Eca 109/Adr cells were more irregular in shape and size, and no microvilli on the Eca 109/Adr cell surface was observed, only slight expression of mdr 1 gene was detected by PCR analysis. The intracellular GSH level was higher than Eca 109 cells. The growth rate of Eca 109/Adr cells was slower and the colony forming ability was lower than the parental cells. Conclusion: The Eca 109/Adr cells were multidrug resistant cells.