耐5-氟尿嘧啶的人结肠癌HT-29/5-FU细胞株的建立及其生物学特性  

Establishment and biological characterization of 5-fluorouracil-resistant human colon cancer HT-29/5-FU cell line

作　　者：邱婷婷[1] 项晓军[1] 雷婉[1] 郝超[2] 张凌[1] 冯苗[1] 余锋[1] 黎军和[1] 熊建萍[1] 

机构地区：[1]南昌大学第一附属医院肿瘤科 [2]南昌大学泌尿外科研究所,江西南昌330006

出　　处：《细胞与分子免疫学杂志》2015年第3期328-332,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81160281)

摘　　要：目的建立耐5-氟尿嘧啶人结肠癌细胞株(HT-29/5-FU)并观察其生物学特性。方法采用5-FU持续接触浓度递增法诱导;观察细胞形态变化和克隆形成率;MTT法测定生长曲线和药物敏感性;Western blot法分析胸苷酸合成酶(TS)、二氢嘧啶脱氢酶(DPD)的表达情况;流式细胞术检测细胞周期和凋亡。结果 HT-29/5-FU的耐药指数为3.59;与HT-29细胞相比,HT-29/5-FU细胞形态发生改变、生长缓慢、克隆形成率降低,TS和DPD高表达;S期细胞比例增多,更能耐受5-FU诱导的细胞凋亡。结论 HT-29/5-FU细胞株生物学性状的改变可能是5-FU化疗耐药机制的重要组成部分,有助于阐明肿瘤潜在耐药机制和逆转肿瘤耐药。Objective To establish 5-fluorouracil （5-FU）-resistant human colon cancer HT-29 cell line （HT-29/5-FU） in vitro and observe its biological properties. Methods The HT-29/5-FU cell line was established by continuously exposing the HT-29 cells to ascending doses of 5-FU. The morphology and colony formation rate were detected. The growth curve and the chemosensitivity of HT-29/5-FU cell line were determined by MTT assay. Western blotting was used to analyze the expressions of thymidylate synthase （TS） and dihydropyrimidine dehydrogenase （DPD）. Cell cycle and apoptosis were measured by flow cytometry. Results The resistance of HT-29/5-FU cells to 5-FU was 3.59-fold greater than that of HT-29 cells. The morphology of HT-29/5-FU cells differed from that of HT-29 cells. Compared with HT-29 cells, HT-29/5-FU cells showed remarkable reduction of cell proliferation and colony formation, higher expressions of TS and DPD, higher percentage of cells in the S phase, and stronger ability of resistance to apoptosis induced by 5-FU, Conclusion The biological characters of HT-29/5-FU cell line may play an important role in 5-FU-resistant mechanisms, which may also contribute to elucidate the potential mechanisms of tumor drug resistance as well as resistance reversal.

关 键 词：结肠肿瘤 HT-29细胞 氟尿嘧啶 抗药性 细胞培养 

分 类 号：R735.35[医药卫生—肿瘤] R392.5[医药卫生—临床医学]