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

卷:

21卷

期数:

2015年04期

页码:

292-295

栏目:

基础医学

出版日期:

2015-07-15

文章信息/Info

Title:

Establishment and identification of cell lines with stable expression of FOXQ1 in MDA-MB-231-luc

文章编号:

1006-8147（2015）04-0292-04

作者:

蔡 隽

（天津医科大学肿瘤医院肿瘤研究所，国家肿瘤临床医学研究中心，天津市“肿瘤防治”重点实验室，乳腺癌防治教育部重点实验室，天津300060 ）

Author(s):

CAI Jun

（1. Cancer Institute and Hospitai,Tianjin Medical University,National Clinical Research Center of Cancer, Tianjin Key Laboratory of Cancer Prevention and Therapy,  Key Laboratory of Breast Cancer Prevention and Therapy,Tianjin Medical University, Ministry of Education, Tianjin 300060, China)

关键词:

FOXQ1; 过表达; 乳腺癌; 迁移; 侵袭

Keywords:

FOXQ1;  over-expression;  breast cancer;  migration;  invasion

分类号:

R737.9

DOI:

-

文献标志码:

A

摘要:

目的：建立稳定过表达FOXQ1乳腺癌细胞系，为进一步研究FOXQ1在体内乳腺癌转移、血管生成、抗凋亡机制，以及以FOXQ1为靶点开发抗肿瘤药物提供细胞研究模型。 方法：采用脂质体转染的方法将真核表达载体FOXQ1转染至MDA-MB-231-luc细胞，经G418筛选及克隆化培养，获得稳定过表达FOXQ1蛋白的细胞系。通过RT-PCR、Western blot和免疫荧光对细胞系中FOXQ1的表达和定位进行鉴定。通过Transwell迁移侵袭等实验，观察过表达FOXQ1后对231-luc细胞EMT相关蛋白表达、231-luc细胞迁移侵袭等生物学特性的影响。 结果：获得过表达FOXQ1的稳定细胞系，迁移侵袭实验表明该细胞系恶性化程度明显高于野生型。 结论：成功建立稳定过表达FOXQ1的乳腺癌细胞系，FOXQ1在细胞核中高表达，进而增加EMT间质标志物Fn1、Vim的表达，增强231-luc的迁移侵袭能力。

Abstract:

Objective: To investigate the contribution of temozolomide easter (TMZ-HE) to the proliferation and apoptosis of glioma LN-18 cells in vitro compared with temozolomide (TMZ). Methods: Methyl thiazolyl tetrazolium (MTT) assay was used to measure the effects on the proliferation of LN-18 cells cultured with graded concentrations of TMZ and TMZ-HE for 24, 48, 72 and 96 h. The long term effect was measured by colony formation assay. Hoechst 33342 and PI fluorescence staining were used to observe the cellular morphological changes and cell necrosis. The AnnexinV-fluoresce in isothiocyanate (FITC) and propidium iodide (PI) assay by flow cytometer were applied to detect the apoptosis induced by TMZ and TMZ-HE. Western blot detected the expressing of resistant protein MGMT in LN-18 after treatment with TMZ and TMZ-HE. Results: TMZ-HE obviously suppressed the proliferation of LN-18 cells in a concentration-and time-dependent manner compared with TMZ. TMZ-HE induced more cellular apoptosis of the LN-18 cells in a concentration-dependent manner. TMZ-HE could block the expressing of MGMT in LN-18 earlier than TMZ. Conclusion: TMZ-HE inhibits proliferation and induced apoptosis by blocked resistant protein MGMT in LN-18 cells. These effects are much stronger than TMZ.

参考文献/References:

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相似文献/References:

备注/Memo

备注/Memo:

作者简介 蔡隽（1989-），女，硕士在读，研究方向：生物化学与分子生物学；E-mail:caijun9865@126.com。

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更新日期/Last Update: 2015-07-16