结直肠癌多药耐药细胞株的建立及其特性的初步探讨  

Establishment of Multidrug Resistant human colorectal cell line(LOVO/5-FU) and its biological characters

作　　者：姚学清[1] 林锋[1] 卿三华[2] 

机构地区：[1]广东省人民医院胃肠外科,广州510080 [2]南方医科大学南方医院普外科,广州510515

出　　处：《广东医学》2006年第6期793-795,共3页Guangdong Medical Journal

基　　金：广东省自然科学基金资助项目(编号:5301056);广东省人民医院科技基金项目(20040024)

摘　　要：目的建立人结直肠癌LOVO细胞多药耐药细胞株LOVO/5-FU,并探讨其生物学特性及耐药机制。方法人结直肠癌细胞系LOVO在体外与2.5μg/ml 5-氟尿嘧啶(5-FU)作用,诱导成功LOVO/5-FU耐药细胞株,体外细胞毒性实验观察它们对5-FU,MMC,ADM,DDP,MTX,Arac等6种药物敏感性。用噻唑蓝(MTT)法、光镜及电镜观察两种细胞的形态及结构差异,绘制两种细胞的体外生长曲线。电镜对耐药细胞株LOVO/5-FU及敏感细胞株LOVO进行5-FU诱导细胞凋亡的实验研究。结果成功的建立耐药细胞系LOVO/5-FU,该细胞系对5-FU的耐药指数为16.48,对MMC,DDP和ADM有交叉耐药性,但对MTX,Arac仍保持与LOVO细胞几乎相同的敏感性。在脱离5-FU体外冻存10个月,其耐药性可保持60%。结论本研究成功的建立LOVO/5-FU多药耐药细胞株,其耐药性稳定,是研究大肠癌MDR的理想模型。Objective To investigate the biological characteristics of multidrug resistant variant of human colorectal LO- VO cell line and the MDR mechanism. Methods A multidrug resistant variant of human colorectal cancer resistant to 5 - FU and LOVO/5 - FU was established in vitro by exposing LOVO parent cell to pulse treatment with 2.5μg/ml of the drug over a period of 6 month.IC60 to antitumor agents was measured by MTT method.The morphology was observed by light microscope and electron microscope. The apoptosis caused by 5 - FU was detected by morphology change under light microscope with DNA strand break techniques and in situ tabeling. Results LOVO/5 - FU cell was more resistant to 5 - FU than the LOVO parent cells. LOVO/5 - FU exhibited cross - resistant to 5 - FU, MMC, ADM and DDP, but not to CTX or Arac. Compared to the parent cells, resistant cells had slowed growth rate and lower confluent demity, unlike the LOVO parent cells. Conclusions The changes in biological characteristics of LOVO/5 - FU was probably associated with MDR phenotype.

关 键 词：细胞培养 结直肠癌 多药耐药 细胞凋亡 

分 类 号：R979.1[医药卫生—药品] R735.35[医药卫生—药学]