人胰腺癌5-Fu耐药细胞株的诱导建立及细胞学特性研究

Establishment and Characterization of 5-Fu-resistant Variant of Human Pancreatic Cancer Cell Line

目的建立稳定传代的抵抗5-Fu的人胰腺癌细胞株MiaPaca2-5-Fu,并对细胞生物学特性进行研究。方法逐步增加培养基中5-Fu的浓度,建立对5-Fu耐药的人胰腺癌细胞株MiaPaca2-5-Fu;采用WST法计算出MiaPaca2和MiaPaca 2-5- Fu的半数抑制浓度(IC50)和耐药指敷(RI);检测MiaPaca2和MiaPaca2-5-Fu的生长曲线,计算2个细胞系的倍增时间并进行比较,用流式细胞仪技术检测其细胞周期。结果5-Fu对MiaPaca2和MiaPaca2-5-Fu的IC50分别为(4.29±0.15)μg/mL、(41.55±2.79)μg/mL,RI为9.68(P=0.0019)。根据生长曲线计算出MiaPaca2和MiaPaca2-5-Fu的倍增时间分别为(39.52±0.32)h、(43.27±0.33)h(P=0.0069),2细胞株的G0/G1期、S期、G2/M期均存在显著性差异(P<0.01)。结论成功建立了对5-Fu耐药的人胰腺癌细胞系MiaPaca2-5-Fu,耐药性能明显、稳定,适合于胰腺癌中5-Fu耐药机制的研究。

Objective To establish stable MiaPaca2 cell lines resistant to 5-Fu and describe the characteristics of this resistant variant. Methods Increase the concentration of 5-Fu, which was exposed to human pancreatic cancer cell line MiaPaca2 gradually to get the 5-Fu resistance cell line named MiaPaca 2-5-Fu. The IC50 and resistance index（RI） of MiaPaca2 and MiaPaca 2-5-Fu cell lines were measured by WST assay. Doubling time was calculated and compared based on the growth curve of these two cell lines, and cell cycles were assayed with flowcytometry method. Results The IC50 increased from （4.29 ± 0.15） μg/mL in MiaPaca2 to （41.55 ± 2.79） μg/mL in MiaPaca2-5-Fu after 72 h exposure and the RI was 9.68（P = 0. 0019）. Doubling time of MiaPaca2 and MiaPaca2-5-Fu was （39.52±0.32） h and （43.27±0.33） h respectively （P=0.0069）. The cell cycles of G0/G1 period, S period and G2/M period have a significant difference between these two cell lines（P〈0.01）. Conclusion MiaPaca 2-5-Fu, the 5-Fu resistant phenotype, is a stable and suitable cell model for studying 5-Fu resistant in pancreatic cancer.