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目的:建立人食管癌顺铂(cDDP)耐药细胞系,测定耐药细胞系DNA聚合酶β(polβ)的表达。方法:以临床食管磷癌化疗用药的最低剂量换算终浓度,采用cDDP(10mg/L)反复间歇 24h暴露法作用于食管癌细胞系ECa -109,建立cDDP耐药细胞系ECa 109 /cDDP;MTT法测定药物敏感性,倒置显微镜下观察细胞形态变化,流式细胞仪检测细胞的周期分布,Westernblot测定细胞内DNApolβ表达水平。结果:与ECa- 109细胞比较,ECa -109 /cD DP细胞形态发生了改变,对cDDP的耐药指数为 1. 56;细胞周期发生了变化,S期细胞增多,G0 /G1 期细胞减少,细胞内DNApolβ表达水平增高(P<0. 05或 0. 01)。结论:ECa-109 /cDDP具有耐药表型,其耐药性产生可能与细胞内DNApolβ表达增高有关。
Objective: To establish a cisplatin-resistant human esophageal cancer cell line and determine the expression of DNA polymerase β (polβ) in drug-resistant cell lines. Methods: The esophageal cancer cell line ECa -109 was induced by cDDP (10 mg / L) repeated intermittent 24h exposure at the lowest dose of esophageal cancer chemotherapy. The expression of ECa 109 / cDDP was determined by MTT The drug sensitivity was measured. The morphological changes of cells were observed under inverted microscope. The cell cycle distribution was detected by flow cytometry. The expression of DNApolβ in cells was detected by Western blot. Results: The morphology of ECa-109 / cD DP cells changed compared with ECa-109 cells and the resistance index to cDDP was 1. 56; the cell cycle changed, the number of S phase cells increased, the number of G0 / G1 phase cells decreased, The intracellular DNApolβ expression increased (P <0.05 or 0.01). Conclusion: ECa-109 / cDDP has a drug-resistant phenotype, and its drug resistance may be related to the increase of intracellular DNApolβ expression.