高转移人涎腺黏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 multi drug resistant cell line derived from highly metastatic human salivary gland mucoepidermoid carcinoma cell line MC3

作　　者：缪叶[1] 秦楠[1] 李焰[1] 刘峰[1] 吴军正[1] 

机构地区：[1]西安第四军医大学口腔医学院,710032

出　　处：《实用口腔医学杂志》2009年第3期340-343,共4页Journal of Practical Stomatology

基　　金：国家自然科学基金项目(编号:30672343)

摘　　要：目的:建立人涎腺黏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:采用裸鼠体内成瘤加药诱导然后在体外用恒定浓度间歇法持续加药,建立高转移人涎腺黏液表皮样癌细胞系MC3的耐药细胞系,应用MTT法,细胞计数法,免疫组化,电镜,RT-PCR等方法观察及检测细胞的生物学特性及耐药性。结果:MC3细胞经过5-FU在体内、体外反复作用产生了多药耐药性,耐药细胞系命名为MC3/5-FU。MC3和MC3/5-FU细胞均为单层贴壁生长,局部可见重叠生长,增殖活跃,2种细胞形态及超微结构未见明显差别。MC3/5-FU,MC3细胞的群体倍增时间分别为55h和60h。MC3/5-FU细胞对于5-氟尿嘧啶(5-FU)、博莱霉素(BLM)、长春新碱(VCR)、长春花碱(VBL)、顺铂(CDDP)的耐药倍数分别为13.3、4.2、6.5、6.9、0.7。免疫组化结果显示:P-gp(p蛋白)在MC3/5-FU呈阳性表达,而在MC3则未见表达。PCR结果显示MDR-1基因在MC3/5-FU中表达高于MC3。结论:MC3/5-FU具有多药耐药性。Objective： To establish a multi drug resistant cell line derived from highly metastatic human salivary gland mucoepidermoid carcinoma cell line MC3. Methods： MC3 cells were injected into athymic mice and the mice were given 5-FU by intraperitoneal injection. After in vivo induction the tumor cells were in vitro cultured and exposed to 5-FU at a constant concentration regularly. The 5-FU induced cells were named MC3/5-FU. The characteristic of MC3/5- FU was studied with MTT assay, flowcytometry and transmission electron microscopy. The expression of MDR-1 gene was examined by RT-PCR. Results： The morphology of MC3 and MC3/5-FU ceils were not obviously different. The population doubling time of MC3/5- FU and MC3 was 55 h and 60 h respectively. The resistance index of MC3/5-FU cells to 5-FU, BLM, VCR,VBL and CDDP were 13.3, 4.2, 6.5, 6.9 and 0.7 respectively. PCR electrophoresis assay showed that expression of MDR-1 gene was higher in MC3/5- FU cells than in MC3. MC3/ 5-FU was P-gp positive while MC3 was negative. Conclusion： MC3/5-FU is a multi drug resistant cell line.

关 键 词：多药耐药 高转移人涎腺黏液表皮样癌 5氟尿嘧啶 

分 类 号：R739.8[医药卫生—肿瘤]