氟他胺耐受性前列腺癌细胞系LNCaP-flu的建立

目的建立氟他胺耐受前列腺癌细胞系LNCaPflu,并初步研究其细胞特性。方法在体外氟他胺作用下,连续培养LNCaP细胞,得到可在10-7mol/L氢化氟他胺中稳定生长的LNCaP亚系,LNCaPflu。使用相差显微镜,电子显微镜,流式细胞仪,四甲基偶氮唑蓝(MTT)比色法、放免法和transwell穿膜实验对细胞特性进行鉴定。结果建立氟他胺耐药细胞系LNCaPflu,与普通LNCaP细胞相比,在氟他胺作用下,此细胞株(65.0±1.7)%处于G1期,明显少于对照组的(82.7±2.3)%(P<0.05);前列腺特异性抗原分泌活力无变化,LNCaPflu为(11.25±2.23)ng/L,LNCaP为(10.15±0.86)ng/L(P>0.05);穿膜侵袭能力未受氢化氟他胺明显影响,LNCaP穿膜数为(35.21±4.52)个/高倍视野,LNCaPflu为(26.15±3.69)个/高倍视野,(P>0.05)。结论LNCaP细胞经长时间体外培养可在氟他胺中稳定生长,LNCaPflu细胞可在体外模拟前列腺癌细胞由氟他胺敏感转化为氟他胺不敏感的过程。

Establish of an flutamide insensitive prostate cancer cell line,LNCaP-flu

Objective To establish a new flutamide insensitive subline of androgen responisive prostate cancer cell line LNCaP,and further explore its cellular characters.Methods LNCaP cells were continuously cultured in vitro in the presence of anti-androgen hydroxyflutamide for around 70 generations and a new subline of LNCaP,LNCaP-flu,which could grow in 10~-7 mol/l hydroxyflutamide steadily,was obtained.The biological characters of LNCaP-flu were studied under phase contrast microscopy,electron microscopy,flowcytometer,MTT assay,radioimmunological assay and transwell in vitro invasive potential assay.Results Succeeded in establishing a new flutamide resistant subline of LNCaP,LNCaP-flu.Comparing with average LNCaP cells,it could grow and proliferate steadily in presence of hydroxyflutamide,while still kept the invasive potential and ability of secreting PSA.Conclusion LNCaP-flu cells could grow in presence of flutamide after continuous in vitro culture,and establish of LNCaP-flu could perfectly mimic the procession from flutamide sensitive to flutamide insensitive.