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目的 探讨结肠癌组织中胸苷磷酸化酶 (dThdPase)的表达 ,检定单核 巨噬细胞表达的dThdPase对抗癌药物 5′ 脱氧氟尿苷 (5’ DFUR)的转化调控作用。方法 对 4 0例结肠癌标本进行免疫组织化学染色 ,分析其细胞的dThdPas表达 ,并测定癌组织和周围正常组织的dThdPase活性。应用ELISA法分别检测 4株结肠癌细胞LS174T、CloneA、Colo32 0、MIP10 1和 2株类巨噬细胞THP 1、U937的dThdPase蛋白含量。MTT法测定 4株结肠癌细胞对 5 Fu和 5′ DFUR的半数有效浓度 (IC50 )。把定量5′ DFUR加入培养基中同类巨噬细胞或单核细胞一起培养 2 4h后 ,取其上清液二倍稀释加入大肠癌细胞中测定IC50 有无改变。并在加入定量 5′ DFUR的类巨噬细胞培养液中检测 5 Fu的生成。结果 结肠癌组织中dThdPase活性明显高于周围正常肠组织 ,表达dThdPase阳性细胞主要是肿瘤相关巨噬细胞 (TAM)。 4株结肠癌细胞中仅LS174T检出 0 5单位 /mg的dThdPase蛋白 ;而THP 1和U937则分别为 18 2单位 /mg和 19 3单位 /mg。全部大肠癌细胞对 5′ DFUR的IC50 均明显高于 5 Fu(P <0 0 0 0 1)。但同THP 1、U937或单核细胞一起培养 2 4h后 ,其IC50 明显下降 ,仅相当于原来的 5 4 %～4 1 8% (P <0 0 0 1)。从含定量 5′ DFUR的THP 1、U937的培养?
Objective To investigate the expression of thymidine phosphorylase (dThdPase) in colon cancer tissues and to detect the dThdPase expression in monocyte-macrophage cells and to investigate the regulatory effect of dThdPase on the anticancer drug 5 ’DFUR. Methods 40 cases of colon cancer specimens were immunohistochemically stained for the expression of dThdPas, and the activity of dThdPase in cancer tissue and surrounding normal tissues was determined. The dThdPase protein levels of four colon cancer cell lines LS174T, CloneA, Colo32 0, MIP10 1 and two macrophages THP 1 and U937 were detected by ELISA. The MTT method was used to determine the half effective concentration (IC50) of 4 colon cancer cells against 5 Fu and 5 ’DFUR. The quantitative 5 ’DFUR added to the medium of similar macrophages or monocytes cultured together 24 h later, take the supernatant twice diluted added to colorectal cancer cells to determine IC50 change. 5 Fu production was measured in macrophage-like medium supplemented with quantitative 5 ’DFUR. Results The dThdPase activity in colon cancer tissues was significantly higher than that in the surrounding normal intestinal tissues. The positive expression of dThdPase positive cells was mainly tumor - associated macrophages (TAM). Of the four colon cancer cells, only LS174T detected 0 5 units / mg of dThdPase protein whereas THP 1 and U937 showed 18 2 units / mg and 19 3 units / mg, respectively. The IC50 of all colorectal cancer cells to 5 ’DFUR was significantly higher than that of 5 Fu (P <0 0 0 0 1). However, IC50 decreased significantly after incubation with THP1, U937 or monocytes for 24 hours, which was only 54.4% -41.8% of the original (P <0.01). From the culture of THP 1, U937 containing quantitative 5 ’DFUR?