氟尿嘧啶对结直肠癌RKO细胞SHP2基因表达的影响

王洁; 黄岩; 张川; 马兴元; 常文军; 陆一鸣

doi:10.3969/j.issn.1006-0111.2017.06.008

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氟尿嘧啶对结直肠癌RKO细胞SHP2基因表达的影响

王洁, 黄岩, 张川, 马兴元, 常文军, 陆一鸣

文章导航 > 药学实践与服务 > 2017 > 35(6): 512-515

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]. 药学实践与服务, 2017, 35(6): 512-515. doi: 10.3969/j.issn.1006-0111.2017.06.008

引用本文:

WANG Jie, HUANG Yan, ZHANG Chuan, MA Xingyuan, CHANG Wenjun, LU Yiming. Effect of 5-FU on the expression of SHP2 gene in colorectal cancer cells RKO[J]. Journal of Pharmaceutical Practice and Service, 2017, 35(6): 512-515. doi: 10.3969/j.issn.1006-0111.2017.06.008

Citation:

WANG Jie, HUANG Yan, ZHANG Chuan, MA Xingyuan, CHANG Wenjun, LU Yiming. Effect of 5-FU on the expression of SHP2 gene in colorectal cancer cells RKO[J]. Journal of Pharmaceutical Practice and Service, 2017, 35(6): 512-515. doi: 10.3969/j.issn.1006-0111.2017.06.008

氟尿嘧啶对结直肠癌RKO细胞SHP2基因表达的影响

doi: 10.3969/j.issn.1006-0111.2017.06.008

1.
第二军医大学药学院生化药学教研室, 上海 200433
2.
华东理工大学生物工程学院, 上海 200237
3.
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

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）

Effect of 5-FU on the expression of SHP2 gene in colorectal cancer cells RKO

1.
Department of Biochemistry Pharmacy, School of Pharmacy, Second Military Medical University, Shanghai 200433, China
2.
College of Biological Engineering, East China University of Science and Technology, Shanghai 200237, China
3.
Department of Environmental Hygiene, School of Tropical Medicine and Public Health, Second Military Medical University, Shanghai 200433, China

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。
- 氟尿嘧啶 /
- 蛋白酪氨酸磷酸酶 /
- 结直肠癌 /
- 免疫荧光
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.
- 5-FU /
- SHP2 /
- colorectal cancer cells /
- immunofluorescence

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氟尿嘧啶对结直肠癌RKO细胞SHP2基因表达的影响

doi: 10.3969/j.issn.1006-0111.2017.06.008

1. 第二军医大学药学院生化药学教研室, 上海 200433

2. 华东理工大学生物工程学院, 上海 200237

3. 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

收稿日期: 2017-06-19
修回日期: 2017-07-10

关键词:

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。

English Abstract

Effect of 5-FU on the expression of SHP2 gene in colorectal cancer cells RKO

1. Department of Biochemistry Pharmacy, School of Pharmacy, Second Military Medical University, Shanghai 200433, China

2. College of Biological Engineering, East China University of Science and Technology, Shanghai 200237, China

3. Department of Environmental Hygiene, School of Tropical Medicine and Public Health, Second Military Medical University, Shanghai 200433, China

Received Date: 2017-06-19
Rev Recd Date: 2017-07-10

Keywords:

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.

引用本文: