人胰腺癌吉西他滨耐药细胞系的建立及耐药特性的检测

Establishment and characterization of a gemcitabine-resistant variant of human pancreatic cancer cell line

目的为揭示吉西他滨在胰腺癌中引起耐药的机制,建立了人胰腺癌吉西他滨耐药细胞系并对其细胞生物学特性进行了研究。方法逐步增加培养基中吉西他滨的浓度,在人胰腺癌细胞系SW1990中建立了对吉西他滨耐药的细胞系SW1990-GEM。采用四氮唑蓝(MTT)法和集落形成实验,计算SW1990和SW1990-GEM的半数抑制浓度(IC50)和耐药系数(RI)。比较SW1990和SW1990-GEM的生长曲线并计算出两细胞系的倍增时间。采用MTT法检测SW1990-GEM对几种常用抗肿瘤药物的交叉耐药谱。结果SW1990和SW1990-GEM的IC50分别为(0.428±0.069)μmol/L和(230.53±13.12)μmol/L,RI为537.45(P<0.001)。根据生长曲线计算出SW1990和SW1990-GEM的倍增时间为22.8小时和35.8小时。交叉耐药实验表明,SW1990-GEM对5-FU、表柔比星、丝裂霉素、甲氨蝶呤、紫杉醇、泰素帝、长春新碱和依托泊苷产生交叉耐药,对顺铂、阿糖胞苷无交叉耐药。结论成功建立了人胰腺癌吉西他滨耐药细胞系SW1990-GEM,耐药性能明显、稳定。SW1990-GEM对5-FU、表柔比星、丝裂霉素、甲氨蝶呤、紫杉醇、泰素帝、长春新碱和依托泊苷产生交叉耐药,对顺铂、阿糖胞苷无交叉耐药。

Objective To establish the gemcitabine 2’,2-difluorodeoxycytidine-resistant variant of human pancreatic cell line. Methods Resistance to gemcitabine was established by exposing human pancreatic cancer cell line SW1990 to increasing concentrations of gemcitabine,which is designated as SW1990-GEM.The IC_(50) and resistance index weretested by MTT assay and colony formation test.The growth curve and cell cycle of SW1990 and SW1990-GEM were compared.The cross-resistance profile of SW1990-GEM was also tested. Results The IC_(50) increased from (0.428±0.069)μmol/L in SW1990 to (230.53±13.12)μmol/L in SW1990-GEM as tested by MTT assay at 72 h exposure,with a 537.5-fold elevation (P<0.001).Doubling time of SW1990 and SW1990-GEM was 22.8 h and 35.8 h,respectively as evaluated by growth curve.SW1990-GEM was cross-resistant to 5-fluorouracil,mitomycin C,epirubicin,paclitaxel,doceltaxel,vincristine and etoposide,but not to cytosine arabinoside and cisplatin. Conclusion The gemcitabine-resistant pancreatic cell line SW1990-GEM has been established,which shows a stable resistance to gemcitabine and cross-resistance to a variant of chemotherapeutic agents.