人涎腺粘液表皮样癌耐药细胞系的建立及其生物学特性  

Establishment and characterization of a multidrug resistant cell clone derived from human salivary gland mucoepidermoid carcinoma cell line MEC-1

作　　者：李焰[1] 吴军正[1] 司徒镇强[1] 刘斌[1] 王江华[1] 李峰[1] 李洁[1] 

机构地区：[1]第四军医大学口腔医学院口腔生物学教研室,陕西西安710033

出　　处：《第四军医大学学报》2002年第18期1637-1640,共4页Journal of the Fourth Military Medical University

基　　金：第四军医大学科技创新工程 ( CX99A0 12 )

摘　　要：目的　建立人涎腺粘液表皮样癌细胞系耐药细胞株并观察其生物学特性及耐药性能 .方法　采用浓度递增法长期不间断诱导并经克隆化筛选 ,建立稳定的人涎腺粘液表皮样癌细胞系耐药细胞株 (MEC- 1/ 5 - FU) .应用 MTT法、细胞计数法、软琼脂克隆形成试验、流式细胞术、免疫组化、电镜等方法观察及检测该细胞株的生物学特性、耐药性能及参数 .结果　倒置显微镜观察该耐药细胞和其亲本涎腺粘液表皮样癌细胞系 (MEC- 1)的细胞生长活跃 ,MEC- 1为单层贴壁生长于培养器皿中 ,MEC- 1/ 5 - FU则可见重叠生长 ,两种细胞形态及超微结构未见明显改变 .MEC- 1/ 5 - FU和 MEC- 1细胞的群体倍增时间分别为 2 8.8h和 30 .0 h;克隆形成率分别为4.9%和 1.2 %;细胞周期分布 S期细胞分别为 2 5 .4%和17.1%.MEC- 1/ 5 - FU耐药性能稳定 ,给药剂量已达 1.980μg· L- 1 . 12种化疗药物对 MEC- 1/ 5 - FU和 MEC- 1生长抑制作用的剂量效应参数显示 :耐药细胞诱导作用药物 5 - FU对 MEC- 1/ 5 - FU细胞和 MEC- 1的半抑制浓度 (IC50 )值分别为 6 .310 μg· L- 1和 0 .44 7μg· L- 1 ,耐药倍数为 14.1;药物敏感性试验结果同时可见该耐药细胞株对 VCR,VBL ,PYM,DNR,VP16 ,CODP,PADM等化疗药物的 IC50 较MEC- 1表现不同程度的耐药性 ,AIM To establish and characterize a multidrug resistant cell clone derived from human salivary gland mucoepidermoid carcinoma cell line MEC-1. METHODS MEC-1 cells were exposed to 5-FU at gradually increased concentration from 0.001 μg·L -1 to 1.980 μg·L -1. Then the drug resistant clone was selected by cell cloning technique. The characteristics of the cell clone was studied with MTT assay, cell counting, flow cytometry, clonogenic assay, immunohistochemical staining and electron microscopic observation. RESULTS Resistant cells were capable of sustaining continuous growth in 1980 μg·L -1 of 5-FU for 18 months with 80 passages. The resistance of MEC-1/5-FU cells to 5-FU, VCR, VBL, PYM, DNR, VP16, CODP and PADM was 14.1, 7.1, 7.9, 5.3, 3.6, 3.2, 2.5 and 4.0 times more than that of MEC-1, respectively. The doubling time (h) of MEC-1/5-FU and ME-1 was 28.8 and 30.0, the S phase cells 25.4% and 17.1% and the clonogencity 4.9% and 1.2% respectively. MEC-1/5-FU was P-pg positive, while MEC-1 negative. The morphology of MEC-1/5-FU was similar to that of MEC-1. CONCLUSION MEC-1/5-FU is a multidrug resistant cell clone.

关 键 词：生物学特性 涎腺粘液表皮样癌 细胞株 多药耐药 5-氟尿嘧啶 

分 类 号：R739.87[医药卫生—肿瘤]