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

卷:

25卷

期数:

2019年03期

页码:

210-214

栏目:

基础医学

出版日期:

2019-05-20

文章信息/Info

Title:

Study of the effects of AGR3 on breast cancer cell lines

文章编号:

1006-8147(2019)03-0210-05

作者:

张金曼; 许 乔; 马勇杰; 谷 峰

（天津医科大学肿瘤医院乳腺病理科，国家肿瘤临床医学研究中心，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）

Author(s):

ZHANG Jin-man; XU Qiao; MA Yong-jie; GU Feng

（Department of Breast Cancer Pathology and Research Laboratory， Cancer Institute and Hospital, Tianjin Medical University, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin, Tianjin’s Clinical Research Center for Cancer, Tianjin 300060, China）

关键词:

乳腺癌; AGR3; 增殖; 迁移; 侵袭; 5-氟尿嘧啶; 紫杉醇

Keywords:

breast cancer;  AGR3;  proliferation;  migration;  invasion;  5-FU;  PAC

分类号:

R737.9

DOI:

-

文献标志码:

A

摘要:

目的：构建稳定过表达前梯度蛋白3（AGR3）或者降表达AGR3的乳腺癌细胞系，探讨AGR3在乳腺癌细胞系中的功能。方法：构建AGR3过表达以及降表达的慢病毒载体，在293T细胞中包装成慢病毒，分别感染MDA-MB-231细胞系获得稳定过表达AGR3的细胞株，感染T47D细胞系获得稳定降表达AGR3的细胞株。通过Western blot技术检测细胞中AGR3的表达情况。利用增殖实验检测细胞的增殖速度。通过Transwell实验检测细胞的迁移、侵袭能力。MTT法检测在不同浓度下5-氟尿嘧啶（5-FU）、紫杉醇（PAC）处理48 h后，细胞的存活情况。结果：稳定过表达AGR3的MDA-MB-231细胞系，细胞的增殖速度更快，迁移、侵袭能力明显增强；在5-FU处理下，存活率较对照组低，在PAC处理下，存活率较对照组高。稳定降表达AGR3的T47D细胞系，细胞的增殖速度显著下降。结论： AGR3的表达与乳腺癌细胞的增殖、迁移、侵袭能力呈显著正相关，与细胞对5-FU的敏感性也呈正相关，而与细胞对PAC的敏感性则呈负相关。

Abstract:

Objective: To explore the function of AGR3 in breast cancer cell lines by screening the stable breast cancer cell line expressing AGR3 protein or AGR3-silenced breast cancer cell line. Methods: First lentiviral vector pCDH-p-3×flag-AGR3-HA and pLVX-ZsGreen-shAGR3 were constructed, and then the breast cancer cell lines MDA-MB-231 and T47D were infected with lenvirus collected from 293T cell for stable expression. The protein expression level of AGR3, cell survival ratio, migration and invasion were detected by Western blot, proliferation assay and transwell assay, respectively. The cell viability was tested using the MTT colorimetric method after 48 hours of exposure to 5-FU or PAC at different concentrations. Results: The cell line MDA-MB-231 with stable expression of AGR3 significantly enhanced cell proliferation, migration and invasion abilities. Compared with control group, survival rate for AGR3 overexpression group cell was significantly decreased after the treatment of 5-FU; and the cell survival rate was increased after exposure to PAC. AGR3 knockdown inhibited cell proliferation ability in the T47D cell line. Conclusion: Up-regulation of AGR3 gene expression may significantly increase cell proliferation and metastasis, promote the sensitivity to 5-FU and inhibit the sensitivity of PAC.

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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（81672636）
作者简介 张金曼（1993-），女，硕士在读，研究方向：生物化学与分子生物学；通信作者：谷峰，E-mail：fenggumayo@163.com。

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更新日期/Last Update: 2019-07-03