Nicotinamide induces apoptosis of F9 mouse teratocarcinoma stem cells by downregulation of SATB1 expression.

Abstract

The aim of this study was to decide whether nicotinamide (NA) could induce apoptosis of F9 mouse teratocarcinoma stem cells (MF9) by downregulation of special AT-rich sequence binding protein 1 (SATB1) expression. We used different concentrations of NA (0, 1.5, 2, and 2.5 mmol/L) to treat MF9 cells and analyze SATB1 expression by RT-qPCR and Western blotting; in addition, the cell proliferation was detected in a microplate reader with Cell Counting Kit-8 (CCK-8), and the cell cycle and apoptosis were analyzed using flow cytometry. We found that the expression of SATB1 was decreased significantly in NA-treated groups than in the control group, and its expression level was inversely related to the NA concentration. In addition, CCK-8 analysis showed that NA significantly inhibited the proliferation of MF9 cells, and flow cytometry showed that NA blocked MF9 cells to G1 phase and significantly promoted apoptosis in any treated groups. To confirm the results, we constructed small interference RNA (siRNA) targeting at mouse SATB1 and transferred into MF9 cells. The results indicated that the expression of SATB1 in both mRNA and protein levels was significantly decreased after cells transferred with siRNA sequence for 48 h, the proliferation of MF9 cells was significantly inhibited, and most of MF9 cells were blocked at G1 phase, and the apoptosis rate was increased obviously. The results showed that NA could inhibit the proliferation and induce apoptosis of MF9 cells. These findings might be used as an efficient candidate for teratocarcinoma therapy.