The effect of PRMT 2 gene-targeted microRNA on estrogen-mediated proliferation of breast cancer MCF7 cells

Objective: To construct human protein arginine methyltransferase 2 (PRMT 2) gene-targeted microRNA expressing eukaryotic recombinant, and to investigate its effect on cell proliferation of breast cancer MCF7 cells transfected with PRMT 2 gene-targeted microRNA. Methods: According to the sequence of PRMT2 mRNA, the PRMT2 pre-microRNA was designed and synthesized, and then it was cloned into the GFP tagged pcDNA™6.2-GW/EmGFP-miR vector and transfected into breast cancer MCF7 cells. The integrity of inset fragment was detected by sequencing analysis. The biological activity of recombinant was identified through detecting interference efficiency of PRMT2 microRNA recombinant by indirect immunofluorescence and Western blotting. The proliferation and colony formation of MCF7 cells were measured by crystal violet assay and colony formation assay, and the cell cycle was determined by flow cytometry (FCM). Results: The sequence of inset fragment in microRNA expressing recombinat was correct, and the expression of PRMT2 protein was decreased after transfection. The knockdown of PRMT 2 expression induced by microRNA did not change the growth property of MCF7 cells with no treatment. The microRNA recombinant-transfected MCF7 cells treated with 4-OHT proliferated at the same rate as the control cells, whereas a stronger promotion of the proliferation of the microRNA recombinant-transfected MCF7 cells cultivated with estrogen was observed. The colony formation assay showed that PRMT 2 knockdown induced by microRNA enhanced the estrogen-mediated colony formation ability of MCF7 cells. FCM assay indicated that the percentages of MCF7 cells at G2 phase were increased after PRMT 2 knockdown induced by microRNA (P < 0.05). Conclusion: The PRMT 2 gene-targeted pcDNA™6.2-GW/EmGFP-miR expressing eukaryotic recombinant plasmid can be successfully constructed, and has biological activity in MCF7 cells after stable transfection. PRMT 2 knockdown induced by microRNA can enhance the estrogen-mediated proliferation and colony formation of MCF7 cells, up-regulate the estrogen-stimulated expression of estrogen receptor target genes cyclin D1 and c -myc, and promote the process of cell cycle. Copyright© 2011 by TUMOR.