대한한방종양학회지 (THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY)

대한암한의학회 (Korean Association of Traditional Oncology)
권호 표지

백화사설초 메탄올 추출물에 의한 HL-60 세포고사과정에 있어서의 transcriptional factors 활성변화 연구

Study on Transcriptional Factors Activation Change of HL-60 cell Apoptosis by Hedyotis Diffusa's Methanol Extract

  • Park, Sang-Goo (Dept. of Internal Medicine, College of Oriental Medicine Wonkwang University) ;
  • Lee, Ji-Hyun (Dept. of Internal Medicine, College of Oriental Medicine Wonkwang University) ;
  • Moon, Gu (Dept. of Internal Medicine, College of Oriental Medicine Wonkwang University) ;
  • Moon, Suk-Jae (Dept. of Internal Medicine, College of Oriental Medicine Wonkwang University) ;
  • Won, Jin-Hee ;
  • Park, Lae-Gil
  • 발행 : 2000.12.30
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초록

Objective : Hedyotis diffusa has been used as an anticancer agent for several decades in oriental medicine. We test whether the methanol extract of the herb affects transcriptional activation factors including $NF-{\kappa}B$ and AP-1. Methods : 1. HL-60 cells were treated with various concentrations(from 200 to $50{\mu}g/ml$) of methanol extract and $H_2O$ extract($200{\mu}g/ml$)of hedyotis diffusa, After 48h later, the cells were tested for viability by MTT assay. 2. The HL-60 cells were treated with $200{\mu}g/ml$ of methanol extract for the indicated periods. First. Nuclear extracts were isolated and incubated with oligonucleotide probe of $NF-{\kappa}B$ and AP-1. Second. Nuclear extracts were isolated and reacted with p50, p65. c-rel pan-Jun, c-Jun, JunB. JunD antibody on ice for 30min. Finally The cell lysates were prepared and analyzed by western blotting using anti-Fas, anti-FasL and anti-p53 antibody. Results : 1. The methanol extract decreases the viability of human lymphoid origin leukemia HL-60 cells in a dose-dependent manner. 2. $NF-{\kappa}B$ is rapidly activated by the addition of the methanol extract, reaches a peak at 30min and gradually returns to resting level. We confirm that $NF-{\kappa}B$ is a heterodimer mainly composed of p65 subunit with c-Rel. 3. Transcriptional activation of AP-1 is detected at 30min and reaches a maximum at 1hr after stimulation of the cells with the methanol extract. AP-1 is mainly composed with Jur-D and partially Jug-B proteins. 4. the methanol extract of Hedyotis diffusa induces the expression of Fas, Fas ligand and p53 proteins of HL-60 cells in a time dependent fashion. Conclusions : These results suggest that the methanol extract of Hedyotis diffusa exerts anticancer effects to induce the death of human leukomic HL-60 cells via activation of trascriptional factors such as $NF-{\kappa}B$ and AP-1, increase in expression of Fas mediated signalling proteins, and induction of tumor suppressor gene. p53.

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