靶向COL1A1基因的 shRNA对乳腺癌MDA-MB-231细胞体外侵袭与迁移的影响

基础医学与临床 ›› 2013, Vol. 33 ›› Issue (8): 981-985.

靶向COL1A1基因的 shRNA对乳腺癌MDA-MB-231细胞体外侵袭与迁移的影响

余海浪¹,马文丽¹,周珏宇²,孟伟¹,石嵘³,郑文岭²

1. 南方医科大学
2. 南方医科大学基因工程研究所
3. 第一军医大学分子生物学研究所

收稿日期:2012-08-19 修回日期:2012-10-29 出版日期:2013-08-05 发布日期:2013-07-18
通讯作者: 马文丽 E-mail:wenlima1964@163.com
基金资助:
广东省医学科研基金;广东省领军人才专项基金

Effects of shRNA targeting COL1A1 gene on the invasion and migration of human breast cancer MDA-MB-231 cell line in vitro

Received:2012-08-19 Revised:2012-10-29 Online:2013-08-05 Published:2013-07-18

摘要/Abstract

摘要： 目的探讨Ⅰ型胶原α1链（COL1A1）基因沉默对人乳腺癌MDA-MB-231细胞侵袭与转移的影响。方法设计2条shRNA 干扰载体及1 条阴性对照载体, 分别稳定转染乳腺癌MDA-MB-231细胞。用Western blot法筛选抑制效率最高的细胞进行细胞平板集落形成实验、细胞黏附实验、细胞迁移及侵袭实验，观察COL1A1基因对MDA-MB-231细胞粘附、运动及侵袭能力的影响。结果转染pshRNA-COL1A1-2干扰载体的MDA-MB-231细胞COL1A1蛋白表达降低最明显，抑制率达66.98%±2.08%，选择该组细胞作为有效干扰组进行后续实验。pshRNA-COL1A1转染组细胞的集落形成率显著低于空白对照组和阴性质粒pshRNA-scramble转染组(P<0.05)；细胞黏附实验中pshRNA-COL1A1转染组细胞的吸光度值较对照组明显降低(P<0.001)；转染pshRNA-COL1A1质粒组细胞的穿膜细胞数也显著低于空白对照组和阴性质粒pshRNA-scramble转染组(P<0.001)。结论 COL1A1基因沉默可在体外抑制人乳腺癌MDA-MB-231细胞的黏附、侵袭和迁移。

关键词: shRNA, COL1A1, 乳腺癌, 侵袭, 转移

Abstract: Objective To investigate the effects of shRNA-mediated human collagen type 1 alpha 1 （COL1A1）gene silence on the migration and invasion of Breast Cancer Cell Line MDA-MB-231. Methods Two shRNA vectors and one negative control vector were designed and stably transfected into MDA-MB-231 cells. Western blot analysis was used to screen for the group which had the highest inhibitory rate. Monolayer colony formation assay was performed to assess a single cell proliferation ability. The cell adhesion, migration and invasion potencies were observed by cell-matrix adhesion assay, and Transwell assay. Results Cells transfected with pshRNA-COL1A1-2 vector had the stronger inhibition of COL1A1 protein, with the inhibition rate being 66.98%±2.08%, and cells in this group were used for the subsequent experiments. Compared with control group and pshRNA-scramble group, the monolayer colony formation rate decreased obviously(P<0.05). The optical absorbance value of adherent cells and the number of invading and migrating cells which moved across the matrix barrier for pshRNA-COL1A1 group were less that than that of control group and pshRNA-scramble group(P<0.001). Conclusion COL1A1 gene silencing of human MDA-MB-231 cells could inhibit cell matrix adhesion, migration and invasion potencies in vitro.

Key words: shRNA, COL1A1, breast cancer, invasion, metastasis

中图分类号:

R737.9

余海浪马文丽周珏宇孟伟石嵘郑文岭. 靶向COL1A1基因的 shRNA对乳腺癌MDA-MB-231细胞体外侵袭与迁移的影响[J]. 基础医学与临床, 2013, 33(8): 981-985.

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