The Effect Of PRMT2Gene-targeted Interference On Estrogen-Mediated Proliferation Of Breast Cancer Cells

Objective:To interfere the expression of PRMT2gene by using microRNA (miRNA) and toinvestigate its effect on cell proliferation of breast cancer MCF7cells transfected withPRMT2gene-targeted microRNA.Methods:PRMT2gene-targeted miRNA plasmid was constructed and transfected into breastcancer MCF7cells via lipofectamineTM2000. The biological activity of recombinant wasidentified through detecting interference efficiency of PRMT2microRNA recombinantby indirect immunofluorescence and Western blotting. The effect of proliferation ofMCF7cells were measured by crystal violet assay, colony formation of MCF7cells weremeasured by colony formation assay, and the cell cycle was determined by flowcytometry (FCM).Results:1. The sequence of inset fragment in microRNA expressing recombinant wascorrected. The results of indirect immunofluorescence method and western blottingshowed that the expression of PRMT2protein was decreased of recombinant-transfectedMCF7cells.2. Crystal violet showed that the knockdown of PRMT2expression induced bymicroRNA did not change the growth property of MCF7cells with no treatment.Compared with the group transfected control vector, the microRNArecombinant-transfected MCF7cells treated with100nmol/L4-OHT proliferated at thesame rate as the control cells, whereas, a stronger promotion of the proliferation of themicroRNA recombinant-transfected MCF7cells cultivated with10nmol/L E2treatment was observed (P＜0.05).3. Western blotting assay showed that the estrogen-stimulated expression of estrogenreceptor target genes cyclinD1and c-Myc were up-regulated by the microRNArecombinant-transfected MCF7cells.4. The colony formation assay showed that the estrogen-mediated colony formationability of MCF7cells were enhanced when PRMT2knockdown induced by microRNA.5. FCM assay indicated that the percentages of MCF7cells at G2phase wereincreased after PRMT2knockdown induced by microRNA (P＜0.05).Conclusion:1. The PRMT2gene-targeted microRNA expressing eukaryotic recombinant plasmidwas constructed successfully, and then the stable MCF7cell lines of PRMT2knockdownwas constructed successfully.2. The esrogen-mediated proliferation and colony formation of MCF7cells wereenhanced when PRMT2knockdown induced by microRNA.3. The estrogen-stimulated expression of estrogen receptor target genes cyclinD1and c-Myc were up-regulated, and the process of cell cycle was promoted when PRMT2knockdown induced by microRNA.