Effects of silencing APN gene on proliferation, invasion and apoptosis of human glioma U251 cells

Abstract

Objective To determine the effect of aminopeptidase N (APN) on the proliferation, invasion, and apoptosis in human glioma U251 cells and to investigate the possible mechanism. Methods After APN gene expression was silenced in glioma cells U251 by RNAi, its expression at mRNA and protein levels was detected by RT-qPCR and Western blotting, respectively. Then, CCK-8 assay, flow cytometry, wound healing test and transwell assay were used to detect the proliferation ability, cell cycle and apoptosis, migration and invasion activity in the U251 cells. The protein expression of STAT3 signal pathway proteins t-STAT3, p-STAT3, MMP-2 and Bcl-2 was measured by Western blotting. Results Compared with blank control and NC-siRNA cells, the expression of APN was decreased at mRNA and protein levels in APN-siRNA cells (P < 0.01). Silencing of APN gene also resulted in inhibited proliferation ability (P < 0.05), cell cycle arrested at G0/G1 phase, enhanced apoptotic rate (P < 0.01), and decreased migration and invasion abilities (P < 0.01) in U251 cells. What's more, the inhibited expression of p-STAT3 caused the decreased expression of Bcl-2 and MMP-2 (all P < 0.05). Conclusion Silencing of APN could inhibit the proliferation, migration and invasion, and induce apoptosis in glioma U251 cells via STAT3 signaling pathway.

Keywords

• glioma
• aminopeptidase n
• rnai
• stat3
• apoptosis
• invasion