J Korean Acad Oral Health.  2019 Jun;43(2):72-77. 10.11149/jkaoh.2019.43.2.72.

Effect of apoptosis on G361 cells by Cimicifuga rhizoma extract

Affiliations
  • 1Feagle Co., Ltd., Yangsan, Korea.
  • 2Department of Oral Anatomy, School of Dentistry, Pusan National University, Yangsan, Korea.
  • 3Department of Korean Internal Medicine, Pusan National University, Yangsan, Korea. jwhong@pusan.ac.kr
  • 4Department of Dental Hygiene, Kyungnam College of Information & Technology, Busan, Korea. sanglye5@naver.com

Abstract


OBJECTIVES
To investigate whether the cytotoxic effect of Cimicifuga rhizoma extract is associated with cell death in the human keratinocyte (HaCaT) and human melanoma cell lines (G361).
METHODS
Apoptosis induced by Cimicifuga rhizoma extract was confirmed by water-soluble tetrazolium salts-1 (WST-1) assay, immunocytochemistry, and western blot. Additionally, the release of cytochrome c and apoptosis-inducing factor (AIF) was visualized by confocal laser scanning microscopy.
RESULTS
The results showed that Cimicifuga rhizoma extract significantly reduced the viability of G361 cells with half-maximal inhibitory concentration (IC 50) of 200 µg/ml, and the apoptotic process was found to occur via the activation of caspase-3 and caspase-9 pathways. Besides, the release of cytochrome c and AIF was also detected.
CONCLUSIONS
This study suggests that Cimicifuga rhizoma extract causes apoptosis of human melanoma cells through the intrinsic apoptotic pathway.

Keyword

Apoptosis; Cimicifuga rhizoma; Melanoma

MeSH Terms

Apoptosis Inducing Factor
Apoptosis*
Blotting, Western
Caspase 3
Caspase 9
Cell Death
Cell Line
Cimicifuga*
Cytochromes c
Humans
Immunohistochemistry
Keratinocytes
Melanoma
Microscopy, Confocal
Apoptosis Inducing Factor
Caspase 3
Caspase 9
Cytochromes c

Figure

  • Fig. 1 Effects of Cimicifuga rhizoma in HaCaT (A) and G361 cells (B). Cell viability was measuerd by WST-1 assay (P<0.05). a,b,cThe same character indication shows that there is no statistical significance.

  • Fig. 2 Cytochrome c (A) and AIF (B) release visualized by confocal immunofluroescence microscopy.

  • Fig. 3 The total protein lysates were used for SDS-PAGE followed by western blot analysis. Antibodies against PARP-1, DFF 45, caspase-9, and caspase-3 were used.


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