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.

Nicotinamide inhibits melanoma in vitro and in vivo

“…Data reported in the present study support a mechanism likely related to an early alteration of cellular energy metabolism. In fact, we observed that NAM treatment increases NAD + , ATP, and ROS levels at as early as 6 h treatment and leads to a significant increase of cells in G1 phase and relevant depletion of cells in S-G2 phase and a strong increase of cells in apoptosis (sub-G1 phase) at 24 h. Of note, NAM has been shown to induce apoptosis also in mouse teratocarcinoma stem cells [ 54 ] and increased intracellular levels of ATP have been related to cytotoxic effects in other cell types [ 55 ]. All such early effects paralleled the relevant cell number reduction and cell death induction observed at 24 and 48 h treatment.…”