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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

23卷

期数:

2017年06期

页码:

485-488

栏目:

基础医学

出版日期:

2017-11-20

文章信息/Info

Title:

Relationship between T85C mutation in dihydropyrimidine dehydrogenase gene and sensitivity of breast cancer cell to 5-Fluorouracil

文章编号:

1006-8147(2017)06-0485-04

作者:

黄 勇¹; 吕 欣²; 王 伟²; 马勇杰¹

(1.天津医科大学肿瘤医院,国家肿瘤临床研究中心,乳腺癌防治教育部重点实验室,天津市“肿瘤防治”重点实验室,天津市恶性肿瘤临床医学研究中心,天津300060;2.天津医科大学公共卫生学院营养与食品卫生学系,天津 300070)

Author(s):

HUANG Yong¹; Lü Xin²; WANG Wei²; MA Yong-jie¹

(1.Tianjin Medical University Cancer Institute and HospitalNational Clinical Research Center for Cancer, Key Laboratory of Breast Cancer Prevention and Therapy, Tianjin Medical University, Ministry of Education, Key Laboratory of Cancer Prevention and Therapy, Tianjin, Tianjins Clinical Research Center for Cancer, Tianjin 300060, China; 2.Department of Nutrition and Food Hygiene, School of Public Health, Tianjin Medical University, Tianjin 300070, China )

关键词:

乳腺癌;  二氢嘧啶脱氢酶;  突变; 5-氟尿嘧啶;  药物敏感性

Keywords:

breast cancer; DPYD; mutation; 5-Fluorouracil; drug sensitivity

分类号:

R737.9

DOI:

-

文献标志码:

A

摘要:

目的：探讨二氢嘧啶脱氢酶基因（DPYD）T85C位点突变对乳腺癌细胞5-氟尿嘧啶（5-FU）敏感性的影响。方法：从肝脏组织中提取mRNA，扩增得到野生型DPYD编码序列。利用定点突变技术，获得T85C定点突变的DPYD编码序列。将GFP、野生型和突变型DPYD序列，连入PCDH-CMV-MCS-EF1-Puro载体。转染上述三种质粒到MDA-MB-231细胞中，获得稳定表达的细胞株。MTT法检测不同浓度5-FU（0、1、5、10、50μg/mL）处理24h后，细胞存活情况。结果：酶切验证三种重组质粒，观察到目的序列。测序验证定点突变质粒，突变位点发生T到C的改变。相比对照组，突变型细胞的存活率显著增高（P <0.01），且突变型细胞存活率高于野生型（P <0.05）。结论：DPYD基因突变影响乳腺癌细胞对5-FU敏感性，T85C突变型DPYD细胞对5-FU的敏感性显著降低。

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Abstract:

Objective: To explore the relationship between T85C mutation in dihydropyrimidine dehydrogenase gene (DPYD) and sensitivity of breast cancer cell to 5-Fluorouracil（5-FU）. Methods: Genomic mRNA was isolated from human liver tissue. DPYD coding sequence was amplified as wild type DPYD. The T was transformed into C by site-directed mutagenesis technique at nucleotide position 85 in DPYD. Then the sequences of GFP, wild type and mutation DPYD were inserted into PCDH-CMV-MCS-EF1-Puro vector respectively. The recombinant plasmids were transfected into MDA-MB-231 to obtain stable cells. After 24 hours of exposure to 5-FU at different concentrations（0,1,5,10,50μg/mL）, the cell viability was tested using the MTT colorimetric method. Results: The GFP, wild type and mutation DPYD were observed by restricting double enzyme digestion. The nucleotide of mutation DPYD transformed from T to C at 85 in DPYD coding sequence. Compared with control group, the mutation group cell survival was significantly increased（P <0.01）, and the mutation group cell survival rate was higher than that of wild type group（P <0.05）. Conclusion: The mutation of DPYD influences the sensitivity of breast cancer cell to 5-FU, and the sensitivity of T85C mutation phenotype cell to 5-FU could be significantly reduced.

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备注/Memo

备注/Memo:

基金项目 国家自然科学基金资助项目(81572851)

作者简介 黄勇（1991-），男，硕士在读，研究方向：乳腺癌的发生发展与化疗耐药机制；通信作者：马勇杰，E-mail：yongjiemagu@aliyun.com。

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更新日期/Last Update: 2017-11-14