Basic & Clinical Medicine ›› 2013, Vol. 33 ›› Issue (8): 981-985.

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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

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

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