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目的建立人肺腺癌耐药细胞模型 Anip973/NVB(去甲长春新碱)并鉴定其生物学特性。方法应用人肺腺癌细胞系 Anip973,采用 NVB 逐步增加剂量法,诱导建立耐药细胞模型Anip973/NVB,观察其生长规律;用 MTT 法鉴定抗药性;观察细胞形态和超微结构;流式细胞技术检测其细胞周期分布;高效液相色谱法测定细胞内 NVB 的浓度变化。结果经 MTT 法鉴定 Anip973/NVB 细胞较 Anip973细胞的 NVB 半数致死浓度(IC_(50))增大21.81倍。两细胞系的倍增时间无明显差异。光镜及电镜下观察到,两细胞系结构变化较大。经流式细胞仪测定 Anip973/NVB 细胞,S 期细胞减少(P=0.035)而 G_0～G_1期细胞增多(P=0.014);高效液相色谱法检测发现 Anip973细胞内药物浓度明显高于 Anip973/NVB。结论 Anip973/NVB 细胞是一个明确的多药耐药细胞模型,具有耐药细胞的基本生物学特性。
OBJECTIVE: To establish Anip973 / NVB (norvincristine) resistant human lung adenocarcinoma cell model and identify its biological characteristics. Methods The human lung adenocarcinoma cell line Anip973 was used to establish the drug-resistant cell model Anip973 / NVB by increasing the dose of NVB gradually. The growth of the cells was observed by MTT assay. The morphology and ultrastructure of the cells were observed. Flow cytometry The cell cycle distribution was detected by HPLC. The concentration of intracellular NVB was measured by HPLC. Results MTT assay showed that Anip973 / NVB cells increased 21.81 times the median lethal concentration (IC 50) of Anip973 cells. There was no significant difference in doubling time between the two cell lines. Light and electron microscopy showed that the structure of the two cell lines changed greatly. Anip973 / NVB cells were detected by flow cytometry. The number of cells in S phase (P = 0.035) and G_0 ~ G_1 phase increased (P = 0.014). The drug concentration in Anip973 cells was significantly higher than that of Anip973 / NVB. Conclusion Anip973 / NVB cells are a clear multidrug resistant cell model with the basic biological characteristics of drug-resistant cells.