氟尿嘧啶对结直肠癌RKO细胞SHP2基因表达的影响
投稿时间:2017-06-19  修订日期:2017-07-10  点此下载全文
引用本文:王洁,黄岩,张川,马兴元,常文军,陆一鸣.氟尿嘧啶对结直肠癌RKO细胞SHP2基因表达的影响[J].药学实践杂志,2017,35(6):512~515
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作者单位E-mail
王洁 第二军医大学药学院生化药学教研室, 上海 200433  
黄岩 第二军医大学药学院生化药学教研室, 上海 200433  
张川 华东理工大学生物工程学院, 上海 200237  
马兴元 华东理工大学生物工程学院, 上海 200237  
常文军 第二军医大学热带医学与公共卫生学系环境卫生教研室, 上海 200433  
陆一鸣 第二军医大学药学院生化药学教研室, 上海 200433 bluesluyi@sina.com 
基金项目:国家自然科学基金资助项目(30500093,81274162);国家科技部"重大新药创制"专项(2009ZX09103-690);上海市科委生物医药领域科技支撑项目(16431904400);上海市教委科研创新项目(14ZZ077)
中文摘要:目的 探究氟尿嘧啶(5-FU)对结直肠癌RKO细胞SHP2基因表达的影响。方法 培养对数生长期的人结直肠癌细胞RKO,5-FU处理48 h后,采用免疫荧光和Western免疫印迹观察SHP2的蛋白表达水平;将能够特异性抑制SHP2表达的siRNA(siSHP2)转染RKO细胞,5-FU处理48 h,CCK8测定细胞吸光度(A)值,流式检测细胞凋亡,观察RKO对5-FU的敏感性变化及5-FU诱导细胞凋亡的影响。结果 结直肠癌细胞RKO在5-FU处理48 h后,其细胞核中SHP2的蛋白表达水平明显增高;siSHP2转染后,RKO细胞对5-FU的敏感性降低,且降低了5-FU诱导的细胞凋亡率。结论 5-FU可能通过影响SHP2的表达,促进RKO细胞的凋亡,发挥抑癌作用。
中文关键词:氟尿嘧啶  蛋白酪氨酸磷酸酶  结直肠癌  免疫荧光
 
Effect of 5-FU on the expression of SHP2 gene in colorectal cancer cells RKO
Abstract:Objective To investigate the effect of 5-FU on the expression of SHP2 gene in colorectal cancer cells RKO.Methods The human colorectal cancer cell line RKO of logarithmic growth phase was cultured 48 h with 5-FU. The expression of SHP2 was observed by immunofluorescence and Western blotting; siSHP2 which specifically inhibits the expression of SHP2 was transfected into RKO cells and cultured 48 h with 5-FU. The cell absorbance (A) values were measured with CCK8 and apoptosis was determined by flow cytometry. The sensitivity of RKO to 5-FU and the effect of 5-FU on apoptosis of RKO cells were observed.Results The expression of nuclear protein of SHP2 was improved remarkably after colorectal cancer cell RKO was cultured 48 h with 5-FU. The sensitivity of RKO cells to 5-FU and the cell apoptosis rate induced by 5-FU were decreased after siSHP2 transfection. Conclusion 5-FU exerts anti-cancer activity possibly due to its promoting the apoptosis of RKO cells by influencing the expression of SHP2.
keywords:5-FU  SHP2  colorectal cancer cells  immunofluorescence
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