摘要
目的建立胰腺癌耐药细胞株,鉴定并分析耐药细胞株的生物学特性。方法选择人胰腺癌细胞株SW1990,采用5-氟尿嘧啶(5-FU)浓度梯度递增法建立胰腺癌耐药细胞株SW1990/FU;电镜观察超微结构;染色体核型分析对比耐药和亲本细胞株在遗传学上的变化;细胞计数法绘制细胞生长曲线,计算倍增时间;流式细胞仪(FACS)测定细胞周期;MTT法计算4种药物的耐药指数(RI);酶联免疫吸附试验(ELISA)测定肿瘤标记物CEA和CA19-9浓度;BHLB/c-m裸鼠种植瘤实验观察肿瘤形成的时间和大小。结果胰腺癌耐药细胞株具有特殊的形态学变化,明显区别于亲本细胞。染色体核型分析显示,SW1990/FU具有特异的18号染色体四倍体核型。与亲本细胞株相比,SW1990/FU生长缓慢(P<0.05),倍增时间延长(P<0.05),大部分肿瘤细胞集中在G0-G1期。MTT结果表明,SW1990/FU对5-FU、阿霉素(ADM)、丝裂霉素(MMC)和吉西他滨(GEM)的耐药指数分别为132.7、3.2、1.5和4.7。ELISA实验表明,在细胞数和培养时间相同的条件下,SW1990/FU分泌的CEA和CA19-9明显高于SW1990(P<0.05)。裸鼠种植瘤实验显示,第4周时耐药细胞株肿瘤直径为(1.5±0.30)cm,明显高于亲本细胞株的(0.8±0.15)cm(P<0.05)。结论5-FU浓度梯度递增法诱导建立的胰腺癌耐药细胞株具有独特的形态学、生物学及遗传学特点。SW1990/FU的建立可作为体外细胞实验模型用于探索在胰腺癌化疗中产生获得性耐药的机制,并为寻找克服肿瘤耐药的方法提供实验基础。
Objective To establish a 5-fluorouracil （5-FU）-resistant pancreatic adenocarcinoma （PAC） cell strain, and to investigate its biological characteristics. Methods The PAC cell strain SW1990 was selected into a muhidrug-resistant cell strain stepwise with 5-FU, one of the most common drugs used in PAC chemotherapy, for 12 months and subsequently named SW1990/FU. The cell strain was characterized in terms of morphology, biology, and cross-resistance to adriamycin（ADM）, mitomycin-C （MMC）, and ge＇mcitabine. BHLB/c-m nude mice tumor growth and CEA and CA19-9 levels were analyzed. In addition, karyotyping and FACS analysis were performed in SW1990/FU and SW1990. Results The SW1990/FU cell strain was 132.7 times more resistant to 5-FU than the parental SW1990 cells, and exhibited cross-resistance to other agents. Compared to the parental cells, SW1990/FU cells exhibited a smaller growth rate, delayed cell-doubling time, and specific changes in chromosomes 18. Tumor diameters in multidrug resistance and parental cells inoculated in in vivo experiments were （1.5±0.30） cm and （0.8±0.15） cm, respectively. Conclusions Morphological adaptation and intracellular changes can be induced by drug challenge in PAC cells. SW1990/FU may be used as an experimental system for the search to overcome drug resistance and to elucidate possible mechanisms of acquired drug resistance in human pancreatic cancer.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2005年第5期592-596,共5页
Acta Academiae Medicinae Sinicae
基金
国家自然科学基金(30371389)~~
