Purpose: We’ve previously reported that PRDX2 plays an oncogenic role in colon cancer. Cell apoptosis was investigated with Annexin V/PI staining and flow cytometry and visualized with Hoechst/PI staining assay. All statistical analysis was performed with SPSS 17.0 software. Results: PRDX2 knockdown led to 210 up-regulated genes and 16 down-regulated genes in HCT116 cells. We also found that DNM3 expression was up-regulated following PRDX2 silencing in HCT116 and HT29 K03861 cells. In colon cancer patients, DNM3 was down-regulated and showed a significant association with pathologic grading. DNM3 overexpression inhibited cell proliferation and induced apoptosis in HCT116 and HT29 cells. Cell migration and invasion were also down-regulated in DNM3 overexpressing colon cancer cells, which might be due to the inhibition of MMP9 proteolytic activities. After thorough investigation of the potential mechanism involved, we hypothesized that DNM3 overexpression induced activation of the mitochondrial apoptosis pathway and inhibition of the AKT pathway. Conclusion: These data suggest that DNM3 is down-regulated in colon cancer, serving as a tumor suppressor. Our study provides new sights into the prognostic value and therapeutic application of DNM3 in colon cancer. test. em P /em 0.05 was considered significant statistically. Results DNM3 can be negatively controlled by PRDX2 in human being cancer of the colon cells To research the mRNA manifestation profile of PRDX2 in human being cancer of the colon, RNA-Seq by PossionDis was performed to detect considerably differentially indicated genes (|log2 collapse modification| 1 & Q worth 0.001) in PRDX2 silenced HCT116 cells. The outcomes indicated that PRDX2 knockdown resulted in 210 up-regulated genes and 16 down-regulated genes (Shape 1ACC). Ten differentially indicated genes were chosen (Desk S1) to become Cdx1 additional validated by K03861 qRT-PCR evaluation. We discovered that in the chosen applicant genes, the manifestation of DNM3 was highly controlled by silencing of PRDX2 (Shape S1). Consequently, DNM3 was chosen for further tests. Then, we looked into the proteins manifestation of DNM3 and PRDX2 in cancer of the colon K03861 lines HT29, SW480, HCT116 and SW1116 (Shape 1D). Protein manifestation of PRDX2 was highest in HT29 cells, second highest in HCT116 cells and most affordable in SW1116 cells (Shape 1D). Oddly enough, DNM3 manifestation was most affordable in HT29 cells and highest in SW1116 cells (Shape 1D). To help expand validate the relationship between PRDX2 and DNM3 proteins manifestation, PRDX2 was knocked down in HCT116 and HT29 cells (with higher PRDX2 manifestation) and K03861 up-regulated in SW480 and SW1116 cells (with lower PRDX2 manifestation). As demonstrated in Shape 1E, PRDX2 knockdown resulted right into a considerably increased manifestation of DNM3 in both HCT116 and HT29 cells ( em P /em =0.001, em P /em =0.0002) weighed against NC cells, while PRDX2 overexpression significantly reduced the DNM3 manifestation in both SW480 and SW1116 cells weighed against the NC group ( em P /em =0.0002, em P /em =0.0002). These suggested that DNM3 could be a downstream effector of PRDX2. Open in another window Shape S1 Manifestation of selected genes. Notes: After PRDX2 was silenced, the mRNA expression of selected genes was examined using RT-PCR. * em P /em 0.05. All experiments were performed in triplicate. Abbreviation: NC, unfavorable control. Open in a separate window Physique 1 The mRNA expression profile of PRDX2 knockdown in HCT116 cells and DNM3 is usually a downstream protein of PRDX2. (ACC) PRDX2 siRNA was conducted and introduced into HCT116 cells. The mRNA expression profile was determined by RNA-Sequencing. The volcano plot (A) and heatmap (B) of different RNA expression profile, and the number of differently expressed genes (C). (D) The protein expression of PRDX2 and DNM3 was detected by Western blot in HT29, SW480, HCT116 and SW1116 cells. (E) The effects of PRDX2 knockdown or PRDX2 overexpression on DNM3 expression were tested by Western blot. The intensities of protein bands were quantified by Image J software. GAPDH was used as an internal control. * em P /em 0.05. All experiments were performed in triplicate. Abbreviations: DNM, Dynamin; DEGs, differently expressed genes;.