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Archive > Volume 20 Issue 3 > 2013,20(3):295-300. DOI:10.3872/j.issn.1007-385X.2013.03.007 Prev Next
Basic Research
Construction of miRNA-126 eukaryotic expression vector and its inhibitory effect on proliferation and migration of breast cancer cells
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Abstract:
Objective:To construct a recombinant eukaryotic expression vector encoding miRNA-126 and to explore its effect on proliferation and migration of mouse breast cancer 4T1 cells. Methods: Sense and antisense oligonucleotides of miRNA-126 were designed and synthesized respectively. The eukaryotic expression vector pcDNA6.2-miR-126 was constructed and transiently transfected into 4T1 cells in vitro. The transfection efficiency was observed under a fluorescent microscope. The expression level of miRNA-126 in 4T1 cells was determined by real-time PCR. The proliferation and colony formation ability of 4T1 cells were detected by MTT assay and colony formation assay. The migration of 4T1 cells in vitro was determined by scratch assay. Results: pcDNA62-miR-126 eukaryotic expression vector was successfully constructed and miRNA-126 was effectively expressed in 4T1 cells. Compared with that in empty plasmid transfected group (pcDNA6.2-Ctrl) , the proliferation capacity of 4T1 cells in vitro was obviously decreased in pcDNA6.2-miR-126 transfected group after transient transfection for 72 hours （\[0.30±0.03\] vs \[0.51±0.04\], P<0.05). The migration capacity of 4T1 cells in pcDNA6.2-miR-126 transfected group was also significantly inhibited after transfection for 48 hours (\[8.17±2.30\] vs \[28.33±2.16\], P<0.05). Conclusion:Overexpression of miRNA-126 may inhibit the proliferation and migration of breast cancer 4T1 cells.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No. 81260398), the International Cooperation Foundation of Guizhou Province (No.10C315), and the Collaborative Foundation of Guizhou Province (No. 2011-51)
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