• Molecules and Cells
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• 제39권3호
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• pp.217-228
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• 2016

To generate a biobetter that has improved therapeutic activity, we constructed scFv libraries via random mutagenesis of several residues of CDR-H3 and -L3 of hu4D5. The scFv clones were isolated from the phage display libraries by stringent panning, and their antiproliferative activity against HER2-positive cancer cells was evaluated as a primary selection criterion. Consequently, we selected AH06 as a biobetter antibody that had a 7.2-fold increase in anti-proliferative activity ($IC_{50}$: 0.81 nM) against the gastric cancer cell line NCI-N87 and a 7.4-fold increase in binding affinity ($K_D$: 60 pM) to HER2 compared to hu4D5. The binding energy calculation and molecular modeling suggest that the substitution of residues of CDR-H3 to W98, F100c, A101 and L102 could stabilize binding of the antibody to HER2 and there could be direct hydrophobic interactions between the aromatic ring of W98 and the aliphatic group of I613 within HER2 domain IV as well as the heavy and light chain hydrophobic interactions by residues F100c, A101 and L102 of CDR-H3. Therefore, we speculate that two such interactions were exerted by the residues W98 and F100c. A101 and L102 may have a synergistic effect on the increase in the binding affinity to HER2. AH06 specifically binds to domain IV of HER2, and it decreased the phosphorylation level of HER2 and AKT. Above all, it highly increased the overall level of p27 compared to hu4D5 in the gastric cancer cell line NCIN82, suggesting that AH06 could potentially be a more efficient therapeutic agent than hu4D5.

• 산업식품공학
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• 제15권1호
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• pp.1-5
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• 2011

Codonopsis lanceolata L. (Campanulaceae) has long been used in traditional Korean medicine to treat bronchitis, cough, and inflammatory diseases, however, the efficacy of anti-tumor activities remains to be defined. In this study the effects of Codonopsis lanceolata (C. lanceolata) on proliferation, migration and adhesion in lung (A549, H1299) and ovarian cancer (SKOV-3) cells were investigated. To assess and compare the pharmacological effects and production places of C. lanceolata, the ethanolic extracts of C. lanceolata from different places in Korea (Hongseong, Yecheon, Yeongwol, Yanggu, Gangjin, and Hoengseong) were prepared. The extract from Hoengseong county did have only marginal anti-proliferative activity in all the cell lines tested, however, other extracts had little or no effect on cell proliferation. The extracts from Hongseong, Gangjin or Hoengseong county had partial anti-migratory activity in lung cancer cells, but not in ovarian cancer cells. In addition, the extract from Hoengseong county had partial anti-adhesive activity in ovarian cancer cells, however, other extracts did not affect cell adhesion in both lung and ovarian cancer cells. Taken together, these findings provide the first description of anti-tumor efficacy of C. lanceolata from different production places in Korea, and suggest that C. lanceolata from Hoengseong county may have therapeutic potential in lung and ovarian cancers.

• 한방안이비인후피부과학회지
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• 제22권3호
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• pp.47-62
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• 2009

Glenditsia spina (GS) can resolve carbuncle, relive swelling, dispel wind and destroy parasites. For these reasons, GS has been widely used as dermatologic agent clinically. In this study, the specific pathways of anti-proliferative effect of GS on human derived melanoma cells were identified. The molecular profile was measured using microarray technique to identify up- or down-regulated genes in SK-MEL-2 cell line. Pathway analysis was done by listing percentage of pathway involvement, and the represented pathways were obtained from KEGG. The transcription factor binding sequences were obtained by Transfac database. By the promoter analysis, up-regulated genes by GS were mainly associated with MAPK, Regulation of actin cytoskeleton, Wnt, Focal adhesion and Long term potentiation pathway. Down-regulated genes by GS were mainly associated with MAPK and Antigen processing and presentation pathway. And some of the transcription factors like Sp1 and NF-Y in up-regulated genes and Oct-1 in down-regulated genes by GS also identified.

• The Korean Journal of Physiology and Pharmacology
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• 제13권4호
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• pp.309-313
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• 2009

Spontaneous hypertensive rats (SHR) are an established model of genetic hypertension. Vascular smooth muscle cells (VSMC) from SHR proliferate faster than those of control rats (Wistar-Kyoto rats; WKY). We tested the hypothesis that induction of heme oxygenase (HO)-1 induced by aprotinin inhibits VSMC proliferation through cell cycle arrest in hypertensive rats. Aprotinin treatment inhibited VSMC proliferation in SHR more than in normotensive rats. These inhibitory effects were associated with cell cycle arrest in the G1 phase. Tin protoporphyrin IX (SnPPIX) reversed the anti-proliferative effect of aprotinin in VSMC from SHR. The level of cyclin D was higher in VSMC of SHR than those of WKY. Aprotinin treatment downregulated the cell cycle regulator, cyclin D, but upregulated the cyclin-dependent kinase inhibitor, p21, in VSMC of SHR. Aprotinin induced HO-1 in VSMC of SHR, but not in those of control rats. Furthermore, aprotinin-induced HO-1 inhibited VSMC proliferation of SHR. Consistently, VSMC proliferation in SHR was significantly inhibited by transfection with the HO-1 gene. These results indicate that induction of HO-1 by aprotinin inhibits VSMC proliferation through cell cycle arrest in hypertensive rats.

• KSBB Journal
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• 제28권6호
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• pp.415-422
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• 2013

Gamma irradiation (${\gamma}IR$) is widely used for radiotherapy as a treatment of cancer cells although it has a risk to damage normal cells. Inflammation is regarded as one of side effects of ${\gamma}IR$ while the effect of low dose of ${\gamma}IR$ on inflammation has not been researched well. Here, we investigated the inflammatory responses of low dose of ${\gamma}IR$ on murine spleen cells. It was evaluated if ${\gamma}IR$ affected the mitogen-induced lymphocyte proliferation, the regulation of various inflammatory cytokines (IFN-${\gamma}$, IL-2, IL-17, IL-4, IL-10), and the involvement of Ikaros and MAPK/NF-${\kappa}B$ medicated mechanism. Exposure of $^{137}Cs-{\gamma}IR$ below 2 Gy decreased the lymphocytes proliferative response to mitogens (LPS, ConA) except at the lowest dose, 0.05 Gy. IL-17, IL-2 and IL-4 mRNA increased at 0.5 and 2 Gy, but not altered at 0.05 Gy. IL-10, anti-inflammatory cytokine, increased only at 0.05 Gy. In regard to intracellular signaling, p-JNK, p-p38 and p-$I{\kappa}B{\alpha}$ were not changed, whereas the activation of ERK and Ikaros increased at the lowest dose. These results suggest that exposure of ${\gamma}IR$ less than 0.5 Gy (or below 0.05 Gy) has beneficial effects as a radiation hormesis on immune function.

• 동아시아식생활학회지
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• 제24권1호
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• pp.20-27
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• 2014

Our study is investigated the effects of Nelumbo nucifera root extract on HT-29 colon cancer cells. The anti-proliferative effect of 70% ethanol extract from Nelumbo nucifera root on HT-29 colon cancer cells was identified based on cell viability, Hoechst 33342 nuclear staining, apoptosis analysis, Western blotting and RT-PCR analyses. In our study, Nelumbo nucifera root extract inhibited the growth of HT-29 colon cancer cells in a dose-dependent manner. Concomitant activation of the mitochondria-dependent apoptotic pathway of HT-29 colon cancer cells by Nelumbo nucifera root extract occurred via modulation of Bax and Bcl-2 expressions, which activated cleavage of caspases-3 and -9. The findings of this study indicate that Nelumbo nucifera root extract induces apoptosis in HT-29 colon cancer cells, and this phenomenon is occurs via the death receptor-mediated and mitochondria-mediated apoptotic pathways.

• 대한한방부인과학회지
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• 제19권1호
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• pp.155-165
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• 2006

이 연구는 MCF-7 인간 유방암 세포주에 대한 귀출파징탕(歸朮破癥湯) 추출물의 증식억제효과 세포독성효과 세포사 유발효과를 확인하기 위하여 이루어졌다. MCF-7 인간 유방암 세포주는 Dulbecco's modified Eagle's medium/F12 (DMEM/F12)에 10% fetal bovine serum(FBS;Gibco) 와 항생제를 가하여 만든 배지를 이용하여 배양하였고, MCPF-7 세포를 96- well plate에 접종한 후 다양한 농도(0~2000g/ml)의 귀출파징탕(歸朮破癥湯)이 든 배지로 처리하고 다양한 시간(48, 96, 192)동안 배양하여 현미경으로 관찰하고 각각 MTS assay kit를 이용하여 세포생존율을 측정하였다. 세포독성은 Sulforhodamine B assay 방법을 이용해 측정하였고 세포사 과정에서 MCF-7세포에서의 caspase 활성화를 측정하기 위해 Western blotting을 수행하여 poly ADP ribose polymerase(PARP)의 절단을 확인하였다. 실험결과 귀출파징탕(歸朮破癥湯) 추출물에 의한 세포성장 및 독성효과는 시간 및 농도에 비례하는 것으로 나타났고 세포고사과정에서 작용하는 caspase의 전 기질인 PARP 절단량이 귀출파징탕(歸朮破癥湯) 처리 농도와 시간에 비례하여 증가하였다. 이것은 caspase-3가 MCF-7 세포의 성장을 억제하는데 중요한 역할을 수행함을 의미한다. 따라서 귀출파징탕(歸朮破癥湯)은 다양한 기전에 의해서 유방암 세포에 대한 억제효과를 가진다는 것을 인식할 수 있다.

• Archives of Pharmacal Research
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• 제25권4호
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• pp.480-484
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• 2002

Costunolide has been reported to be a cytotoxic and chemopreventive agent. This work investigated the mechanism of the anti proliferative effect of costunolide and determined that it induced differentiation of the human leukemia cell line HL-60. Costunolide exhibited a potent antiproliferative activity against HL-60 cells. It was also found to be a potent inducer of differentiation in human leukemia derived HL-60 cells through the examination of differentiation markers, as assessed by the reduction of nitroblue tetrazolium, the increase in esterase activities and phagocytic activity, morphology change and the expression of CD14 and CD66b surface antigens. These results, accompanied by a decline in the expression of c-myc protein, suggest that costunolide induces differentiation of human leukemia cells to granulocytes and monocytes/macrophages lineage.

• 한국해양바이오학회지
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• 제14권2호
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• pp.93-101
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• 2022

Hizikia fusiforme, a type of brown algae, is widely used in Asian cuisine. It has been reported to have various pharmacological effects. In this study, the effects of the ethanol extract from H. fusiforme (EAHF) on the proliferation of human colon carcinoma cells were investigated. The effect on the survival of human hepatocarcinoma and colon carcinoma cells was examined, and results revealed that the anti-proliferative effects of EAHF were higher in colon carcinoma cells than in hepatocarcinoma cells. The inhibition of proliferation of HT29 colon carcinoma cells by EAHF treatment was closely related to the induction of apoptosis. EAHF treatment also increased caspase activity and poly(ADP-ribose) polymerase degradation, induced mitochondrial dysfunction, altered Bcl-2 family protein expression, and increased the rate of cytochrome c released from the mitochondria into the cytoplasm. Furthermore, the production of reactive oxygen species (ROS) was markedly stimulated by EAHF treatment, and when ROS production was blocked, EAHF-induced cytotoxicity was significantly attenuated. These results indicate that the anticancer activity of EAHF in HT29 colon carcinoma cells was induced by ROS-dependent mitochondrial impairment. While EAHF exhibited potent anticancer activity in colon carcinoma cells in this study, further studies on the active components of EAHF and their efficacy should be performed.

• Journal of Nutrition and Health
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• 제53권2호
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• pp.111-120
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• 2020

본 연구는 인간 유래 유방 암세포 MDA-MB-231를 대상으로 CA에 의한 세포사멸, 세포 이동 및 침윤 효과를 조사하였다. 암세포의 증식 억제 효과는 CA 농도 의존적으로 증식률이 감소하는 것을 확인하였다. CA에 의한 apoptosis 양성 세포를 확인하기 위해 DAPI stain를 진행한 결과, CA 농도 의존적으로 죽은 세포를 확인하였다. MDA-MB-231 세포에서 CA에 의한apoptosis marker 단백질 발현 증가와 ROS production증가를 확인할 수 있었다. 또한 CA에 의한 MDA-MB-231의 세포 이동률이 유의적으로 감소하는 것을 확인하였다. 마지막으로 세포의 이동과 전이 능력 또한 CA를 처리한 군에서 통계적으로 감소하는 것을 확인하였다. 본 연구 결과를 통해서 CA의 암세포 증식률 억제, 세포사멸 증가, 그리고 세포 이동 및 전이 억제 효과가 나타나는 것을 확인했으며, 이 결과를 통해서 향후 유방암에 대한 항암제로 개발될 수 있는 가능성을 가지고 있는 것으로 사료된다.