Construction of eukaryotic expression vectors of short hairpin RNA for MMP1

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Medical Journal of the Chinese People Armed Police Forces ›› 2012, Vol. 23 ›› Issue (2) : 6-6.

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Construction of eukaryotic expression vectors of short hairpin RNA for MMP1

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Abstract

Objective To construct eukaryotic expression vectors of RNA interference specific to matrix metalloproteinase 1（MMP1）. Methods Genome sequences of MMP1 gene were retrieved from Genbank and cDNA that was capable of coding expression of shRNA (small hairpin RNAs) was designed for MMP1 gene. The cDNA was synthesized and inserted into plasmid pWH1, and the recombinant pWH1-MMP1 expression vector was identified using enzyme cutting. Then, pWH1-MMP1 expression vector was transfected into salivary adenoid cystic carcinoma ACC-M cells. Finally, the expression of MMP1 in ACC-M cells transfected with pWH1-MMP1 expression vector was evaluated. Results Compared with ACC-M cells and ACC-M cells transfected with plasmid pWH1, the ACC-M cells transfected with pWH1-MMP1 expression vector showed a low mRNA and protein expression of MMP1. Conclusions The constructed MMP1 shRNA expression vector can block the expression of MMP1 in salivary adenoid cystic carcinoma cells, potentially facilitating further studies on the significance of MMP1 in the proliferation, metastasis and experimental treatment of salivary adenoid cystic carcinoma.

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Construction of eukaryotic expression vectors of short hairpin RNA for MMP1[J]. Medical Journal of the Chinese People Armed Police Forces. 2012, 23(2): 6-6

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