Identification Of PRMT2-targeting Genes And Its Mechanisms Controlling CCND1Expression In Breast Cancer

Objective: To indentify PRMT2-targeting genes in breast cancer cells, inspect theeffect of PRMT2downregulated on related protein expression, and to investigate themolecular mechanism of PRMT2regulating the expression of CCND1.Methods: To indentify PRMT2-targeting genes by Agilent oligo microarray. Tovalidate the microarray data, nine upregulated genes and nine downregulated genes(Table2) were chosen for real-time PCR. Inspect the effect of PRMT2downregulatedon related protein expression and investigate the molecular mechanism of PRMT2regulating the expression of CCND1by western blot.Results:1. Iidentification of target genes by cDNA microarray showed that a total of2173PRMT2-targeting genes. Most of these differentially genes are involved intranscription regulation, cell adhesion, immune responses, cell growth, proliferationand carcinogenesis. CCND1is one of important target genes of PRMT2.2. The results of real-time PCR showed that five of nine are upregulated genes andeight of nine was downregulated genes, consistent with the results of the microarrayanalysis. Interestingly, CCND1, a key modulator in cell cycle and proliferation, wasfound to be one of the downstream targets of PRMT2.3. Western blotting assay showed that PRMT2gene expression knockdown caused toa statistically significant enhancement of E2-induced CCND1protein expressionlevels, consistent with the results from the microarray and real-time PCR analysis.4. Western blotting assay results suggest that PRMT2inhibits CCND1expressionthrough the suppression of Akt/GSK-3β signaling in breast cancer cells Conclusions:1. CCND1is one of the downstream targets gene of PRMT2, which was suppressedby PRMT2in gene and protein level.2. PRMT2downregulates CCND1expression via the suppression of Akt/GSK-3βsignaling in breast cancer cells.