Mpz gene suppression by shRNA increases Schwann cell apoptosis in vitro

Abstract

We investigated the effects of short hairpin RNA (shRNA) on myelin protein zero (MPZ) gene expression in Schwann cells (SCs) in vitro and determined the effects of the MPZ gene suppression on the survival of SCs. The MPZ-specific shRNA was introduced into a lentiviral vector for expression under the U6 promoter, and the viral vector-based shRNAs were used to infect cultured SCs. The efficiency of MPZ knockdown was analyzed by real time-PCR (RT-PCR) and western blotting. Flow cytometric analysis and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) were used to determine the cell cycle and the amount of apoptosis of SCs. We found that MPZ shRNAs significantly inhibited the expression of the MPZ gene and induced SC apoptosis in vitro. These results provided interesting experimental evidence for understanding the mechanism of demyelinating neuropathies caused by MPZ gene malfunction.