人肺腺癌吉西他滨耐药细胞系的建立及其特性

背景与目的:临床前和临床研究显示吉西他滨对实体瘤有较好的疗效,尽管已有卵巢癌和红白血病吉西他滨耐药细胞系的报道,但肺癌吉西他滨耐药细胞系尚未见报道。我们建立了人肺腺癌吉西他滨耐药细胞系,并对其细胞生物学特性进行研究。方法:逐步增加培养基中吉西他滨的浓度,建立了对吉西他滨耐药的人肺腺癌细胞系A549-Gem。采用四甲基偶氮唑蓝(MTT)法和集落形成实验,计算出A549和A549-Gem的半数抑制浓度(IC50)和耐药系数(RI)。比较A549和A549-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A549-Gem对几种常用抗肿瘤药物的交叉耐药谱。结果:吉西他滨对A549和A549-Gem的IC50分别为(6.56±1.19)μmol/L和(921.09±225.27)μmol/L,RI为140.52(P=0.0195)。集落形成实验的RI为132.95。根据生长曲线计算出A549和A549-Gem的倍增时间为29.7h和36.4h。交叉耐药实验表明,A549-Gem对长春新碱(VCR)和足叶乙甙(VP-16)的RI分别为54.38和6.18(P<0.01),对阿霉素(ADM)、顺铂(DDP)、阿糖胞苷(Ara-C)和紫杉醇(Taxol)无交叉耐药。结论:成功建立了人肺腺癌吉西他滨耐药细胞系A549-Gem,耐药性能明显、稳定,非常适合用于肺癌中吉西他滨耐药的研究。A549-Gem对VCR、VP-16产生交叉耐药,对ADM、DDP、Ara-C和Taxol无交叉耐药

Development and characterization of a gemcitabine-resistant variant of human lung adenocarcinoma cell line A549

BACKGROUND & OBJECTIVE: Gemcitabine (2',2-difluorodeo- xycytide) has antitumor activity in both experimental and clinical treatment of solid tumors. Although resistance to gemcitabine in ovarian cancer cell line and erythroleukemic cell line was described, there was no report on the lung cancer resistant variant. In order to elucidate the mechanism by which gemcitabine induce resistance in lung cancer, we have established the resistance to gemcitabine in human lung adenocarcinoma cell line A549 and described the characteristics of its resistant variant. METHODS: Resistance to gemcitabine was established by exposing A549 cells to increasing concentration of gemcitabine, which was designated as A549-Gem. The IC(50) and resistance index(RI) were tested by MTT assay and colony formation test. The growth curve and cell cycle of A549 and A549-Gem were compared. The cross-resistance profile of A549-Gem was also tested. RESULTS: The IC(50) increased from 6.56+/-1.19 micromol/L in A549 to 921.09+/-225.27 micromol/L in A549-Gem as tested by MTT assay at 72h exposure, the RI was 140.52 (P=0.019 5).The RI of colony formation test was 132.95. Double time of A549 and A549-Gem were 29.7 h and 36.4 h, respectively, as evaluated by the growth curve. A549-Gem was cross-resistant to vincristine and etoposide(54.38-fold and 6.18-fold)(P< 0.01), but not to adriamycin, cisplatin, cytarabine, and paclitaxel. CONCLUSION: A549-Gem, the gemcitabine resistant phenotype, is stable and suitable for the study of gemcitabine resistance in lung cancer. A549-Gem is cross-resistant to vincristine and etoposide, but not resistant to adriamycin, cisplatin, cytarabine and paclitaxel.