类巨噬细胞增强5'-脱氧氟尿苷抗结直肠癌细胞活性  

Enhancement of anticancer effect of 5′deoxy 5 fluorouridine by macrophage like cells on colorectal carcinoma cells

作　　者：张继民[1] 刘明姬[2] 沟井贤幸[3] 椎叶健一[3] 佐々木严 松野正纪[3] 

机构地区：[1]广州医学院第二附属医院胃肠外科,510260 [2]中山大学黄埔医院麻醉科 [3]日本东北大学医学部第一外科

出　　处：《中华胃肠外科杂志》2004年第3期218-221,共4页Chinese Journal of Gastrointestinal Surgery

基　　金：教育部留学人员科研启动基金犤教外司留(2002)247号犦

摘　　要：目的探讨类巨噬细胞表达的胸苷磷酸化酶(dThdPase)能否增强细胞内激活抗癌药物5'-脱氧氟尿苷(5'-DFUR)抗结直肠癌细胞作用。方法应用ELISA法分别检测结直肠癌细胞系LS174T、CloneA、Colo320、CX-1、LOVO、MIP101和类巨噬细胞系THP-1、U937的dThdPase蛋白含量。采用MTT分析,分别测定出氟尿嘧啶(5-FU)和5'-DFUR在6株结直肠癌细胞的半数有效浓度(IC50)。把5-FU或5'-DFUR同THP-1或U937细胞一起培养24h,其上清液2倍稀释后加入结直肠癌细胞中行MTT分析,测定IC50有无改变。并在培养THP-1或U937的培养基中加入定量5'-DFUR后检测5-FU的生成量。结果6株结直肠癌细胞中仅LS174T检出0.5U/mg、LOVO检出8.9U/mg的dThdPase蛋白,其他4株未检出。而THP-1和U937的dThdPase蛋白含量则分别为18.2U/mg和19.3U/mg。5'-DFUR对全部癌细胞的IC50高于5-FU的14.5～94.4倍(P<0.01)。5-FU和5'-DFUR同THP-1(或U937)细胞一起培养24h后,5-FU对6种癌细胞的IC50无明显改变(P>0.05),而5'-DFUR的IC50则分别下降到原来的5.4%～41.8%(P<0.001)。并从加入400μmol5'-DFUR的THP-1和U937的培养液中分别检出40.2μmol和32.9μmol的5-FU。结论6株结直肠癌细胞基本无dThdPase活性,不能在细胞内转化5'-DFUR为5-FU。类巨噬细胞表达的dThdPase可以转化5'Objective To investigate whether dThdPase expressed in macrophage like cells can modulate the anticancer effect of 5′ DFUR on colorectal cancer cell lines. Methods dThdPase levels were detected in 6 kinds of colorectal cancer cells, LS174T, Clone A, Colo320, CX 1, LOVO, MIP101, and 2 macrophage like cells, THP 1, U937 by ELISA. After the drug sensitivities of each colorectal cancer cell line both to 5′ DFUR and 5 FU were estimated by MTT assay, THP 1 and U937 cells were incubated in the media containing 5 FU or 5′ DFUR for 24 hours, respectively. Then the supernatants were added into incubated cancer cells at a 2 fold serial dilution and MTT assay was carried out on each of 6 colorectal cancer cell lines. Finally, the amount of generated 5 FU was measured by HPLC from the supernatants in which THP 1 or U937 cells were incubated for 24 hours with 400 μmol of 5′ DFUR. Results The dThdPase levels were 0.5 U/mg and 8.9 U/mg in LS174T and LOVO lines, 18.2 U/mg and 19.3 U/mg in THP 1 and U937 respectively, but could not be detected in other 4 colorectal cancer cell lines. All colorectal cancer cells were resistant to 5′ DFUR compared with 5 FU. The mean IC50 values of 5′ DFUR were 15 to 94 times higher than those of 5 FU. Incubation with THP 1 or U937 significantly enhanced the antitumor effect of 5′ DFUR and IC50s of 5′ DFUR decreased to 5.4%～41.8%of those untreated lines (P< 0.0001), while no effect on IC50s of 5 FU was observed (P >0.05). The level of 5 FU were 40.2 μmol and 29.5 μmol detected in the media containing 5′ DFUR treated with THP 1 and U937 cells, respectively. Conclusions 5′ DFUR can not be converted to 5 FU in 6 colorectal carcinoma cell lines, because of little dThdPase level in those cells. dThdPase expressed in macrophage like cells can convert 5′ DFUR to 5 FU and release it, resulting in an enhancement of anticancer effect on carcinoma cells.

关 键 词：类巨噬细胞 5′-脱氧氟尿苷 结直肠癌 癌细胞活性 抗癌药 检测 

分 类 号：R735.3[医药卫生—肿瘤]