摘要
目的 :探讨氟尿嘧啶 (5 fluorouracil,5 Fu)诱导体外人结肠癌细胞株 (HIC/S)细胞凋亡与Rb蛋白表达之间的影响。方法 :应用MTT比色法和琼脂培养克隆形成法检测 5 Fu对HIC/S细胞增殖抑制作用 ;采用DNA断片法和末端标记法 (TUNEL)观测 5 Fu对HIC/S细胞凋亡的诱导效应。使用S P免疫组化技术对HIC/S细胞Rb蛋白表达状态进行再证实并用Westernblot法对HIC/S细胞Rb蛋白作半定量分析。结果 :1.5 Fu(0 16～10 0 0 0 μg/ml)显著抑制HIC/S细胞增殖和克隆形成能力 ,呈剂量相关性 (P <0 0 5～ 0 0 0 1)。 2 .5 Fu(12 0 0 μg/ml)处理HIC/S细胞 48小时后出现细胞凋亡的典型“梯形条带”图谱 ,细胞呈现细胞凋亡的特征性蓝紫色颗粒。 3.S P免疫组化染色证实HIC/S细胞Rb蛋白表达阳性 ;5 Fu(12 0 0 μg/ml)处理HIC/S细胞 48小时后 ,可使HIC/S细胞内Rb蛋白表达量增高 (曲线下面积占总面积百分比 :47 0 %VS 2 1 2 % )。结论 :5 Fu能有效地诱导HIC/S细胞凋亡 ;5 Fu诱导HIC/S细胞凋亡与促进HIC/S细胞内Rb蛋白表达增高相关。
Objective:To investigate the roles that 5 fluorouracil (5 Fu) plays in the expression of Rb gene and inducing apoptosis of human colorectal cancer line(HIC/S) cells cultured in vitro Methods:The MTT colorimetric assay and the agar cultured colony formation method were used to examine the growth inhibition of HIC/S cells treated with 5 Fu,and DNA fragmentation method and TUNEL staining were used to observe the apoptosis induced by 5 Fu in HIC/S cells Expression of pRb of HIC/S cells were proved again by Streptavidin-Peroxidase(S P) immunohistochemical method and western blot analysis of Rb protein isolated from HIC/S cells Results:1 5 Fu (0 16～100 00 μg/ml) inhibited significantly proliferation and colony formation of HIC/S cells, which is in dose dependente a manner (P< 0 05～0 001) 2 HIC/S cells treatad with 5 Fu (12 00 μg/ml) for 48 hours showed DNA fragment ladder and bluepurplish grain which are characteristric for apoptosis by DNA agrose gel electrophoresis and TUNEL staining 3 The intracellular expression of pRb in HIC/S cells could promote treated with 5 Fu (12 00 μg/ml) for 48 hours (relative area is 47 0%VS 21 2%) Conclusions:5 Fu induced significant apoptosis of HIC/S cells cultured in vitro 5 Fu induced apoptosis of HIC/S cells was possibly involved in promoting the intracellular expression of pRb
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2000年第5期439-441,共3页
Chinese Journal of Cancer
关键词
RB蛋白
氟尿嘧啶
细胞凋亡
结肠肿瘤
Colorectal neoplasms
Rb protein
5 Fuluorouracil
Apoptosis
