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백화사설초 메탄올 추출물에 의한 HL-60 세포고사과정에 있어서의 transcriptional factors 활성변화 연구
박상구,이지현,문구,문석재,원진희,박래길,Park, Sang-Goo,Lee, Ji-Hyun,Moon, Gu,Moon, Suk-Jae,Won, Jin-Hee,Park, Lae-Gil 대한암한의학회 2000 大韓癌韓醫學會誌 Vol.6 No.1
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.
백화사설초 메탄올 추출물에 의한 HL-60 세포(細胞) 고사과정(枯死過程)에서의 cell cycle 관련인자(關聯因子)의 활성변화(活性變化) 연구(硏究)
한세희,이종범,문구,문석재,원진희,박래길,이종덕,Han, Se-Hee,Lee, Jong-Bum,Moon, Gu,Moon, Suk-Jae,Won, Jin-Hee,Park, Lae-Gil,Lee, Jong-Deok 대한암한의학회 2000 大韓癌韓醫學會誌 Vol.6 No.1
Objectives: Hedyotis diffusa is used to treat cancer in traditional Korea Medicine. So this study was carried out to examine the expression of cell cycle related genes in HL-60 cells undergoing apoptosis by the methanol extract of Hedyotis diffusa. Methods: 1. HL-60 cells were treated with various concentrations (from 200 to $50{\mu}g/ml$)of methanol extract and H20 extract ($200{\mu}g/ml$) of hedyotis diffusa. After 48 h 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. The whole cell lysates were prepared and analyzed by western blotting using anti-p53 antibody. 3. The nuclear extract were prepared and analyzed by western blotting using anti-p21 antibody, anti-p27 antibody, anti-cyclin A antibody, anti-cyclin E antibody and anti-CDK2 antibody. Results: 1. The methanol extract of Hedyotis diffusa induced the death of HL-60 cells in a dose dependent manner. 2. The methanol extract of Hedyotis diffusa makedly decreased the level of p21/Cipl and cyclin A in a time dependent manner. 3. The methanol extract of Hedyotis diffusa markedly increased the level of p27/Kipl and cyclin E in a time dependent manner. 4. The methanol extract of Hedyotis diffusa markedly did not affect the level of CDK2. Conclusions: These results provide evidence that expression of cell cycle related genes in HL-60 cells undergoing apoptosis by the methanol extract of Hedyotis diffusa mainly results from decreased level of p21/Cipl and increased level of p27/Kipl of the cell cycle related genes.
항암제 Cisplatin의 HeLa 세포에 대한 독성기전 연구
최정호 ( Choe Jeong Ho ),조현 ( Jo Hyeon ),이선영 ( Lee Seon Yeong ),오성환 ( O Seong Hwan ),김흥곤 ( Kim Heung Gon ),박래길 ( Park Lae Gil ) 대한산부인과학회 2003 Obstetrics & Gynecology Science Vol.46 No.10
항암제 cisplatin의 항암기전은 DNA cross-linking agent로 인한 genomic DNA의 손상 및 세포 내 활성산소종 생성에 의한 세포손상 등 많은 연구가 이루어지고 있다. 그러나 이러한 노력에도 불구하고 cisplatin에 의해 유도되는 세포 내 신호전달과정을 통한 세포고사는 많은 부분이 알려져 있지 않다. 따라서 본 연구에서는 HeLa 세포에서 cisplatin에 의해 유도되는 세포고사의 신호전달기전, 특히 caspase계 cy Cis-diamminedichloroplatinum Ⅱ (cisplatin) has been reported to induce cell death. However, the mechanism by which cisplatin is induced the apoptosis of cancer cells is still unclear. To evaluate the mechanistic insights of apoptosis by cisplatin, we test
자궁경부암의 Chemotherapeutic Response를 증가시키는 Baicalin Hydrate의 세포 내 신호전달 기전
김병륜 ( Kim Byeong Lyun ),김인숙 ( Kim In Sug ),고경희 ( Go Gyeong Hui ),이제중 ( Lee Je Jung ),김흥곤 ( Kim Heung Gon ),박래길 ( Park Lae Gil ) 대한산부인과학회 2003 Obstetrics & Gynecology Science Vol.46 No.10
Baicalin hydrate는 baicalin이 수화된 것으로 flavonoid계 물질로 PC-SPES의 주요성분이다. PC-SPES는 전립선암 환자와 전립선암을 가진 쥐들에 투여한 결과 환자들의 전립선특이성항원 수치를 낮추고 쥐의 전립선암 종양 크기를 줄인다고 알려져 있다. Baicalin은 항산화작용, 세포독성효과, 역전사효소의 억제에 의한 항바이러스효과 및 혈소판이나 백혈구 등의 골수유래세포, 상피세포, 신경세포 등에 함유되는 12-Lipoxyg Baicalin is flavonoid and major component of PC-SPES. Flavonoids including baicalin have been reported to not only function as anti-oxidant but also cause cytotoxic effect. Baicalin hydrate has been reported to induce cell death, however the mechanism by
Cisplatin에 의한 HeLa세포의 세포고사에 미치는 활성산소종의 영향
고경희 ( Go Gyeong Hui ),김신호 ( Kim Sin Ho ),조해중 ( Jo Hae Jung ),오성환 ( O Seong Hwan ),김흥곤 ( Kim Heung Gon ),박래길 ( Park Lae Gil ) 대한산부인과학회 2003 Obstetrics & Gynecology Science Vol.46 No.12
목적 : 본 연구는 cisplatin이 자궁경부암세포주 HeLa세포에서 세포 고사를 유발하는 신호전달 과정에 세포내 산화제의 축적을 포함하는지의 여부를 알아보기 위하여 수행하였다. 연구 방법 : 자궁경부암 세포주 HeLa세포를 이용하여 체외 실험을 통해 항암치료중 활성산소종의 생성 여부와 항산화 물질을 세포내 투입하여 cisplatin 유래 활성 산소종에 의한 세포사멸 보호효과를 flow cytometry에 의해 측정하였다. 결과 : cisplatin에 Objective : To determine whether oxidants are formed as part of the cisplatin-induced apoptotic process, intracellular markers of oxidative stress were examined. Methods : Apoptotic death of HeLa cells by cisplatin was confirmed by flow cytometry. Results