Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Gene Expression Profiling by Ginsenoside Rb1 in Keratinocyte HaCaT Cells

피부각질세포 HaCaT에서 진세노사이드 Rb1에 의한 유전자 발현 양상

  • Lee, Dong Woo (Department of Biological Science and the Research Institute for Basic Sciences, Hoseo University) ;
  • Kim, Jung Min (Genoplan, Inc. & NAR Center, Inc.) ;
  • Bang, In Seok (Department of Biological Science and the Research Institute for Basic Sciences, Hoseo University)
  • 이동우 (호서대학교 생명과학과 및 기초과학연구소) ;
  • 김정민 (제노플랜코리아(주) 유전체분석팀) ;
  • 방인석 (호서대학교 생명과학과 및 기초과학연구소)
  • Received : 2019.02.26
  • Accepted : 2019.04.20
  • Published : 2019.05.30
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Abstract

We investigated the gene expression patterns and the mechanisms of action of the apoptotic response by microarray analysis of human keratinocyte HaCaT cells treated with ginsenoside Rb1, a saponin of Panax ginseng C. A. Meyer. Genes related to apoptosis, the G2/M transition of the mitotic cell cycle, cell division, mitotic nuclear division, and intracellular protein transport were 2-fold up-regulated in HaCaT cells treated with the ginsenoside Rb1, whereas genes related to DNA repair, regeneration fission, and extracellular matrix organization were 2-fold down-regulated. Apoptosis signaling may be mediated by FAS and PLA2G4A, and pathway analysis indicated that STAT3 might be an upstream regulator of these genes. The activity of FAS and PLA2G4A was verified by qPCR, which showed that FAS was increased about 2-fold in HaCaT cells treated with $10{\mu}g/ml$ of ginsenoside Rb1 for 24 hr, PLA2G4A was increased about twice after 6 hours, and gene expression was increased more than 2-fold after 24 hr. Knockdown of STAT3 with siRNA decreased FAS expression and increased PLA2G4A expression but only FAS was passed from the upstream regulator STAT3. These results indicate that STAT3, which is an upstream regulator, induces apoptosis via FAS during treatment with ginsenoside Rb1.

인삼(Panax ginseng C. A. Meyer)의 사포닌 진세노사이드 Rb1이 처리된 인간 피부각질세포 HaCaT에서 microarray 분석 및 발현이 증가된 세포사멸 반응에 대한 작용기전을 연구하였다. HaCaT 세포에 진세노사이드 Rb1의 처리로 세포사멸, 유사분열 세포주기의 G2/M 전이, 세포분열, 핵분열, 그리고 단백질 수송 등의 작용기전에 관여하는 유전자들이 2 배 이상 발현이 증가된 것으로 나타났으며, DNA 수선, 감수 핵분열, 그리고 세포외기질 체계 등의 작용기전에 관여하는 유전자들은 2 배 이상 발현이 감소된 것으로 나타났다. 특히 세포사멸 신호전달은 FAS와 PLA2G4A를 경유하는 것으로 나타났으며, 이들 유전자의 상위 조절자로 STAT3가 예측되었다. 세포사멸 반응 경유 유전자 FAS와 PLA2G4A의 활성을 qPCR로 확인한 결과, FAS 유전자는 $10{\mu}g/ml$의 진세노사이드 Rb1를 24시간 동안 처리하였을 경우 약 2 배의 발현 증가와, PLA2G4A 유전자는 6시간 처리부터 약 2 배로 증가되어 24시간 동안 처리시 2 배 이상의 유전자 발현이 증가되었다. 한편 STAT3-siRNA를 이용한 knock-down 실험에서 FAS의 발현 감소와 PLA2G4A의 발현 증가로 상위 조절자 STAT3로부터 FAS 만을 경유하는 것을 알 수 있었다. 이상의 결과 진세노사이드 Rb1의 처리에 의해 상위 조절자인 STAT3는 FAS를 경유하여 세포사멸을 유도하는 것임을 알 수 있다.

Keywords

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Fig. 1. The chemical structures of ginsenoside Rb1.

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Fig. 2. Effects of ginsenoside Rb1 on cell viability in HaCaT cells.

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Fig. 3. Cytoprotective effects of ginsenoside Rb1 against H2O2-induced oxidative stress in HaCaT cells.

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Fig. 4. Gene ontology of gisenoside Rb1 by microarray analysis.

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Fig. 5. Gene expression profiles of the genes related to the apoptosis signaling by ginsenoside Rb1.

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Fig. 6. Signaling network of the genes related to the apoptosis signaling.

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Fig. 7. Effect of ginsenoside Rb1 on the gene expressions of FAS (A) and PLA2G4A (B) in HaCaT cells.

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Fig. 8. Effects of knock-down of STAT3 on expression of FAS and PLA2G4A.

Table 1. Primer sequences for qPCR

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Table 2. Gene lists in apoptosis signaling response by Rb1 in the HaCaT cells

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