Obstet Gynecol Sci.  2014 Nov;57(6):501-506. 10.5468/ogs.2014.57.6.501.

Salinomycin inhibited cell proliferation and induced apoptosis in human uterine leiomyoma cells

• Lee HG
• Lee JM
• Shin SJ
• Kwon SH
• Lee GS
• Song CH
• Choi ES
• Cha SD
• Cho CH

• KMID: 2314017
• DOI: http://doi.org/10.5468/ogs.2014.57.6.501

Abstract

OBJECTIVE
The aim of this study was to investigate the anti-proliferative effect of the salinomycin in cell proliferation and apoptosis in primary cultured human uterine leiomyoma cells.
METHODS
Cell viability was measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Caspase-3 activity assay and DNA fragmentation assay were performed to determine the effect of apoptosis. The expression of apoptosis regulatory-related proteins was evaluated by western blot.
RESULTS
The cell viability and proliferation of uterine leiomyoma cells were significantly reduced by salinomycin treatment in a dose-dependent manner. DNA fragmentation assay results showed apoptotic cell death after salinomycin incubation. Salinomycin activated caspase-3, -8, and -9, causing apoptosis in uterine leiomyoma cells. Down-regulation of Bcl-2, XIAP, and FLIP with a concomitant increase in Bax, Fas, and DR5 were observed.
CONCLUSION
These results provided the first evidence that salinomycin induce both intrinsic and extrinsic apoptosis. Therefore, salinomycin may be a promising chemopreventive and therapeutic agent against human uterine leiomyoma.

Keyword

Apoptosis; Extrinsic; Intrinsic; Leiomyoma; Salinomycin

MeSH Terms

Apoptosis*
Blotting, Western
Caspase 3
Cell Death
Cell Proliferation*
Cell Survival
DNA Fragmentation
Down-Regulation
Humans
Leiomyoma*
Caspase 3

Figure

• 

  Fig. 1 Anti-proliferative effect of salinomycin in uterine leiomyoma cells. (A) Cell morphologic changes were observed by phase contrast microscopy. Cells were treated with DMSO (control) or salinomycin (1, 5, 10, and 20 µM) for 24 hours. (B) Cell viability was measured using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Values are ±SD of three measurements *P<0.05.

• 

  Fig. 2 Effect of salinomycin on intrinsic apoptotic pathway related protein expression in uterine leiomyoma cells. Twenty-four hours treated cell extracts were prepared and subjected for immunoblotting analysis. Beta-actin was used as an internal loading control.

• 

  Fig. 3 Effect of salinomycin on extrinsic apoptotic pathway related protein expression in uterine leiomyoma cells. Twenty-four hours treated cell extracts were prepared and subjected for immunoblotting analysis. Beta-actin was used as an internal loading control.

• 

  Fig. 4 Induction of apoptosis after salinomycin treatment in uterine leiomyoma cells. As describe in methodology section cells were exposed to DMSO (control) or salinomycin (5 and 10 µM) for 24 hours. Apoptosis was quantified by (A) caspase-3 activity and (B) enzyme-linked immunosorbent assay. Values are ±SD of three measurements *P<0.05.

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