人乳腺癌细胞凋亡与去磷酸化RB蛋白的关系  

Relationship between cell apoptosis and dephosphorylated RB protein in human breast cancer

作　　者：欧阳学农[1] 王文武[1] 姜浩[1] 

机构地区：[1]南华大学附属第一医院肿瘤科,湖南衡阳421001

出　　处：《中国癌症杂志》2004年第4期306-308,312,共4页China Oncology

基　　金：湖南省卫生厅课题 ( 2 0 0 1 Y5 8)

摘　　要：目的 :研究乳腺癌细胞凋亡与去磷酸化RB蛋白的关系。方法 :以乳腺癌细胞株MCF 7/S(化疗敏感细胞株 )和MCF 7/ADR(耐多柔比星细胞株 )为研究对象 ,用MTT比色法检测乳腺癌细胞的化疗敏感性 ;采用免疫细胞化学法检测多柔比星 (ADR)作用前后RB蛋白的表达状态 ;应用流式细胞术检测ADR诱导乳腺癌细胞凋亡程度。结果 :ADR能抑制敏感细胞MCF 7/S增殖 ,半数抑制浓度 (IC50 )为 0 .12 8μg/ml,而对应的耐药细胞MCF 7/ADRIC50值为 10 .89μg/ml,MCF 7/S细胞较MCF 7/ADR细胞对ADR敏感 86倍。加ADR前两种细胞胞核中的磷酸化RB蛋白均为阳性 ,去磷酸化RB蛋白均为阴性。经不同浓度ADR处理后 ,随着ADR浓度的增加 ,MCF 7/S细胞去磷酸化RB蛋白表达量逐渐增高 ,而MCF 7/ADR细胞去磷酸化RB蛋白表达量无明显增高。流式细胞术检测细胞凋亡和细胞周期 ,结果表明ADR诱导MCF 7/S细胞凋亡作用比MCF 7/ADR强 (P <0 .0 1)。结论 :乳腺癌MCF 7/S细胞凋亡与去磷酸化RB蛋白表达呈正相关 (P <0 .0 5 ) ,而MCF 7/ADR细胞凋亡与去磷酸化RB无明显相关 (P >0 .0 5 )。Purpose:To investigate the relationship between cell apoptosis and dephosphorylated RB protein in human breast cancer. Methods:In our work,human breast cell lines (MCF-7/S,the chemosensitive cell line and MCF-7/ADR,the chemoresistent cell line)were evaluated. Chemosensitivity of two cell lines was evaluated by the MTT colorimetric assay;the expressive levels of dephosphorylated RB protein were detected with immunocytochemistry. Apoptosis rates were determined by flow cytometry(FCM). Results:ADR inhibited proliferation of chemosensitive cell line MCF-7/S ,the 50% inhibition concentration (IC 50 ) was 0.128 μg/ml;And IC 50 of MCF-7/ADR was 10.89 μg/ml. The chemotherapeutic sensitivity of MCF-7/S was more than that of MCF-7/ADR by 86 times . Before treatment with ADR,phosphorylated RB protein was positive in two cell lines,but dephosphorylated RB protein was negative;After treatment of different concentration ADR,when the concentration of ADR was increased,expression of dephosphorylated RB protein elevated accordingly in MCF-7/S,but no significant change in MCF-7/ADR. Apoptosis and cell cycle was detected by FCM assays shows ADR induced apoptosis of MCF-7/S more than MCF-7/ADR(P<0.01). Conclusions:There were positive correlation between apoptosis of MCF-7/S cells and the expression levels of dephosphorylated RB protein,but no significant correlation in MCF-7/ADR(P>0.05).

关 键 词：去磷酸化RB蛋白 乳腺肿瘤 细胞凋亡 多柔比星 

分 类 号：R737.9[医药卫生—肿瘤]