• 대한암한의학회지
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• 제20권2호
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• pp.13-21
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• 2015

Objective : The objective of this study was to investigate the experimental efficacy of anti-tumor activity of the complexed herbal formula, Onbaekwon (OBW), which was derived from the literature of Traditional Korean Medicine, Dongeuibogam. Methods : Nine Cancer cell lines, LoVo, MCF-7, HepG2, AGS, A549, NCI-H69, HL-60, Sarcoma 180, LL/2, were prepared and the cytotoxicity was assessed by 3-(4,5-dimethylthiazol-2yl)-2,5-dephenyl tetrazolium bromide (MTT) assay. Four of them, NCI-H69, HL-60, Sarcoma 180, and LL/2, showed strong cytotoxic activities and they were additionally undergone flow cytometry to find out their effects on apoptosis. ICR male mice were implanted with Sarcoma 180 intraperitoneally and divided into 8 species for each group. Control group was treated with normal saline, positive control group was treated with cyclophosphamide 8mg/kg, and experimental group was treated with OBW 1 g/kg. Results : Among 9 cancer cell lines, NCI-H69, HL-60, Sarcoma 180, and LL/2, expressed less than 0.10 mg/ml of $IC_{50}$ under 0.1~1mg/ml of OBW. NCI-H69, HL-60, Sarcoma 180, and LL/2, showed dose-dependent efficacy of apoptosis. When Sarcoma 180 cancer cell was implanted in ICR male mice and treated with the OBW, they prolonged the median overall survival for 0.8 days, from 17.5 to 18.3. Conclusion : OBW showed strong cytotoxicity to some cancer cells, which are NCI-H69, HL-60, Sarcoma 180, and LL/2, and its apoptotic activity was dose-dependent. OBW prolonged the median survival of mice implanted with Sarcoma 180. Further researches would be expected to support the efficacy of OBW.

• 한국식품영양과학회지
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• 제30권5호
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• pp.921-927
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• 2001

각종 암세포에 대한 번데기동충하초(Cordyceps militaris) 추출물 및 분획물의 세포독성을 규명하기 위하여 암세포로 A549, MCF-7 HeLA, HT1080, Hep3B, KATOIII 및 K562를 이용하였고 그 결과, SRB assay에 의한 1 mg/mL의 에탄올 추출물 농도에서 HT1080, HeLa, Jep3B 그리고 A549는 각각 89.4%, 85.7%, 72.9%그리고 65.5%의 억제효과를 나타내었다. MCF7, HeLa 그리고 HT1080 세포의 경우는 핵산 분획물 1 mg/mL 농도에서 각각 92.9%, 90.3%그리고 97.0%로 다른 분획물보다 현전히 높은 억제효과를 나타냈다. KATOIII세포에 대한 억제효과는 1mg/mL 투여시 에탄올 추출물의 경우 61.5%를 보였고 부탄올과 물 분획물에서 각각 83.7%와 80.4%로 다소 높은 억제효과를 보였다. K562 세포에 대한 시료의 억제효과는 에탄올 추출물(1mg/mL)이 60.5%의 억제율을 보인데 반해 같은 농도에 서 에틸 아세테이트, 부탄올 및 핵산 분획물에서는 각각 88.6%, 806% 및 75.6%로 높은 억제 효과는 양성대조군의 11.2$\pm$0.8에 비하여 10, 20, 40, 80 mg/kg 의 농도에 10.5$\pm$0.8, 8.3$\pm$0.5, 7.8$\pm$0.3, 3.7$\pm$0.6로 8.7, 25.9, 30.4그리고 67.0%의 유의적인 억제효과를 나타내었다. 또한 각각의 분획물에 대한 소핵생성 억제효과는 유전손상물질(MNNG;150 mg/kg, I.P)만 투여한 양성대조군에 비해서 시료 농도를 10, 20, 40 80 mg/kg 처리하였을 경우 동이라한 시료 농도 80 mg/kg 에서 부탄올, 에틸 아세테이트, 물, 그리고 클로로포름 분획물이 각각 3.1$\pm$0.3, 3.7$\pm$0.6, 4.2$\pm$0.3, 4.8$\pm$0.3으로 72.3%, 67.0%, 63.3%그리고 57.1%의 순으로 소핵생성 억제효과를 나타내었으며, 그중 핵산 분획물이 2.8$\pm$0.5의 75.0%로 가장 높은 소핵생성 억제효과를 나타내었다.

• 한국식품조리과학회지
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• 제28권2호
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• pp.175-182
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• 2012

In this study, antibacterial activity and cytotoxicity of the extracts from pericarp and seed of $Vitis$ $coignetiea$, which were extracted with 0.1% HCl-60% ethanol, were analyzed. The antibacterial activity of the extracts was determined by paper disc diffusion method against food spoilage and food-borne pathogens. The pericarp extract showed high antibacterial activity against $Bacillus$ $cereus$, $Escherichia$ $coli$ O157:H7, and $Pseudomonas$ $aeruginosa$, and the seed extract represented the high antibacterial activity against $B.$ $cereus$, $E.$ $coli$ O157:H7, and $Staphylococcus$ $aureus$. The cytotoxicity of the $Vitis$ $coignetiea$ extract against human cancer cells was determined using the MTT assay and SRB assay. The pericarp extract represented strong growth-inhibition activity against G361 and Hep3B cells and the seed extract greatly inhibited the growth of HeLa and G361 cells in the MTT assay. In addition, the pericarp extract displayed a high inhibition activity against the growth of AGS cells and the seed extract greatly inhibited the growth of HeLa, Hep3B, and MCF7 cells in the SRB assay. Especially, the cytotoxicities of the seed extract against HeLa were significantly higher than those of the extract against other cancer cells at all test concentrations. This study demonstrates that the extract from pericarp and seed of $Vitis$ $coignetiea$ possess high antibacterial activity and cytotoxicity.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 춘계학술발표회
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• pp.94-94
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• 2018

The gold (LC-AuNPs) and silver (LC-AgNPs) nanoparticles were rapidly synthesized by fruit extract of Lycium chinense within 1.15 and 25 min respectively in an eco-friendly way. The synthesized nanoparticles confirmed by relevant surface plasmon resonance peaks for gold and silver nanoparticles at 536 and 480 nm, respectively. FE-TEM results revealed that LC-AuNPs were 20-50 nm and LC-AgNPs were 50-100 nm. The maximum distribution of gold, silver elements and the crystallographic nature of synthesized were confirmed using EDX, elemental mapping and XRD. LC-AgNPs showed inhibitory activity against pathogenic microorganisms such as E. coli and S. aureus, whereas LC-AuNPs did not show inhibitory activity. The LC-AgNps nanoparticles exhibited significant cytotoxicity to human breast cancer MCF7 cell line and less cytotoxicity to non-diseased RAW264.7 (murine macrophage) cells whereas LC-AuNps showed minimal toxicity to both cell lines. In-depth research on this rapid, facile and greenery nanoparticles may play a potential role in biomedical applications.

• Journal of Microbiology and Biotechnology
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• 제32권5호
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• pp.551-563
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• 2022

L-asparaginase (E.C. 3.5.1.1) purified from bacterial cells is widely used in the food industry, as well as in the treatment of childhood acute lymphoblastic leukemia. In the present study, the Burkholderia pseudomallei L-asparaginase gene was cloned into the pGEX-2T DNA plasmid, expressed in E. coli BL21 (DE3) pLysS, and purified to homogeneity using Glutathione Sepharose chromatography with 7.26 purification fold and 16.01% recovery. The purified enzyme exhibited a molecular weight of ~33.6 kDa with SDS-PAGE and showed maximal activity at 50℃ and pH 8.0. It retained 95.1, 89.6%, and 70.2% initial activity after 60 min at 30℃, 40℃, and 50℃, respectively. The enzyme reserved its activity at 30℃ and 37℃ up to 24 h. The enzyme had optimum pH of 8 and reserved 50% activity up to 24 h. The recombinant enzyme showed the highest substrate specificity towards L-asparaginase substrate, while no detectable specificity was observed for L-glutamine, urea, and acrylamide at 10 mM concentration. THP-1, a human leukemia cell line, displayed significant morphological alterations after being treated with recombinant L-asparaginase and the IC₅₀ of the purified enzyme was recorded as 0.8 IU. Furthermore, the purified recombinant Lasparaginase improved cytotoxicity in liver cancer HepG2 and breast cancer MCF-7 cell lines, with IC₅₀ values of 1.53 and 18 IU, respectively.

• 생명과학회지
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• 제22권8호
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• pp.1099-1106
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• 2012

체지방감소 효과가 있는 시판 conjugated linoleic acid (CLA)의 색깔을 제거하고 이 색깔의 세포독성에 관한 연구를 하였다. 황갈색의 시판 CLA 제품을 구입하여 crude CLA (c-CLA) 시료로 하였다. c-CLA 시료를 감압증류(10 mmHg-$220^{\circ}C$, 10 mmHg-$235^{\circ}C$, 10 mmHg-$240^{\circ}C$, 20 mmHg-$260^{\circ}C$; 30분)하여 증류된 CLA (distilled CLA; d-CLA) 시료와 증류되지 않고 남아있는 황갈색 CLA (residual CLA; r-CLA) 시료로 분리하였다. 10 mmHg-$220^{\circ}C$에서 증류하여 얻은 d-CLA 시료의 색깔은 L (brightness), a (red/blue), b (yellow/green)로 분석한 결과 무색에 가까웠고 r-CLA 시료는 황갈색이었고, 이들 두 CLA 시료의 CLA 이성체 조성은 변하지 않았다. 따라서 10 mmHg-$220^{\circ}C$에서 얻은 r-CLA 시료의 인체암세포(유방암 MCF-7. 폐암 A-549, 직장암 HT-29, 전립선암 PC-3)와 인체 정상세포(신경모세포 SK-N-SH)에 대한 세포독성을 d-CLA 시료와 비교하였다. 이들 암세포와 정상세포에 r-CLA 시료와 d-CLA 시료 처리 2일 후의 세포독성에는 차이가 없었다. 따라서 본 연구에서 c-CLA 시료에 함유된 색소는 10 mmHg-$220^{\circ}C$로 감압증류 하여 제거할 수 있었고, r-CLA 시료의 세포독성은 d-CLA 시료의 세포독성과 차이가 없었다. 이와 같은 결과는 c-CLA 시료에 함유된 색소는 세포생육에 아무런 영향을 미치지 않고 인체에 아무런 영향을 미치지 않음을 의미한다.

• 동아시아식생활학회지
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• 제15권5
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• pp.542-548
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• 2005

In recent years, propolis has attracted much attention as an useful substance in medicine and functional food, even if it is known as a natural remedy in folk medicine since ancient times. propolis was registered as natural food since 1995 on Korean Food Act by Korean Food and Drug Administration(KFDA). The present study demonstrated the optimization of isolation of crude propolis by ethanol, and tumoricidal effect of pro polis. The optimal concentration of ethanol to separate a high quantity of propolis was $60\%$. The cytotoxic effect of ethanol extracted propolis against various cancer cell lines including murine lymphoma (Sarcoma-180), murine T-lymphoma (YAC-1), human breast carcinoma (MCF-7), human gastric carcinoma (KATOIII) and human hepatocellular carcinoma (Hep3B) and human lung adenocarcinoma (A-549) was observed using SRB and MIT assay. In order to investigate the curative activity by oral administration of propolis on tumor, ICR mice was subcutaneously implanted Sarcoma 180. In 300mg/kg and 600mg/kg propolis administered group, development of implanted tumors was inhibited by $40.9\%\;and\;67.9\%$ at 16th day, respectively. In the same dose of propolis administered group, development of implanted tumors was inhibited more strongly with dose dependent manner. Therefore, these data suggested propolis may show tumoricidal effects. In conclusion, these results indicate that propolis, one of the few natural remedies, can be used as functional food with tumoricidal effects.

• 한국식품과학회지
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• 제32권6호
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• pp.1371-1378
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• 2000

S. typhimurium TA98과 TA100을 이용한 Ames test에서는 아가리쿠스버섯 메탄올 추출물 모두 시료자체의 돌연변이원성은 없는 것으로 나타났다. 아가리쿠스버섯 메탄올 추출물$(200\;{\mu}g/plate)$ 중 TA98 균주를 이용한 항돌연변이 효과를 확인한 결과 직접변이원인 4NQO에 대해 92.4% 그리고 간접변이원인 Trp-P-1과 $B({\alpha})P$에 대해 각각 81.9%와 83.4%의 높은 억제효과를 나타내었다. 또한 TA100 균주에서 4NQO는 94.7%의 가장 높은 억제효과를 나타내었다. MNNG, Trp-P-1 및 $B({\alpha})P$은 각각 87.3%, 89.9% 그리고 92.3%의 억제율을 보였다. 각각의 변이원 물질에 대한 아가리쿠스버섯 분획물 $(200\;{\mu}g/plate)$의 항돌연변이 효과에서는 MNNG와 $B({\alpha})P$에 대해서는 분획물 모두가 80% 이상의 높은 억제율을 보였고, 4NQO는 TA98, TA100 두 균주 모두 에틸 아세테이트 분획물에서 95% 이상의 가장 높은 억제율을 나타내었다. 그리고 Trp-P-1에서는 TA98 균주에서 물층을 제외한 분획물이 높은 억제율을 나타내었고, TA100 균주에 대해서도 분획물 모두가 80% 이상의 높은 억제율을 나타내었다. 각종 암세포에 대한 아가리쿠스버섯 메탄올 추출물의 저해효과는 시료 1 mg/mL 투여시 MCF7 82.9%, A549 86.5%, HT1080 65.5%, Hep3B 84.3%, HeLa 91.9%, KAROIII 88.7% 그리고 K562세포에서는 82.0%의 억제효과를 나타내었다. 인간 정상 간세포 WRL68에 대한 시료 농도에 따른 세포독성효과는 1 mg/mL의 시료를 첨가시 50% 이하의 생육억제율을 나타냄으로써 정상세포에 대해서는 낮은 독성효과를 나타낸다는 사실을 알 수 있었다. 아가리쿠스버섯 분획물들에 대한 억제효과에서는 유방암 세포인 MCF7이 시료를 1 mg/mL 첨가하였을때 물층과 부탄올층을 제외한 분획물에서 90% 이상의 높은 억제효과를 보였다. 또한 에틸 아세테이트 분획물이 다른 분획물에 비해 각각의 암세포에 대해서도 1 mg/mL 첨가시 80% 이상의 높은 억제효과를 보였다. 그러나 물 분획물은 암세포에 대해 상대적으로 낮은 억제효과를 나타내었다.

• Toxicological Research
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• 제14권2호
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• pp.123-127
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• 1998

BRCA1 is a tumor suppresser gene in familial cases of breast cancer. It has been controversial whether the subcellular localization of BRCA1 is located in nuclei or cytoplasm in normal human breast cells. We found that a p220 protein was expressed in Type II Normal human breast epithelial cells (NHBEC) but not in Type I NHBEC in Western blot analysis using the 17F8 (3A2) antibody. Immunostaining using the same antibody revealed positive staining in nuclei, cytoplasm and perinuclei of Type II cells and negative staining in Type I NHBEC. The p220 protein, however, was expressed in SV40 immortalized Type I NHBEC and tumorigenic cells derived from them after x-ray and neu oncogene treatment. The subcelluar localization was mostly cytoplasmic and punctate in the nuclei. The breast carcinoma cell lines, MCF-7 and T47D, also expressed the p220 protein. Using RT-PCR, we observed the expression of BRCA1 mRNA in both Type I and Type II NHBEC. This result indicated that there might be mechanisms involved in post-translational or translational regulation of BRCA1 gene. It is speculated that the absence of BRCA1 protein expression in Type I NHBEC might playa role in their susceptibility to neoplastic transformation.

• Molecules and Cells
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• 제45권6호
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• pp.425-434
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• 2022

The post-translational modification (e.g., phosphorylation) of estrogen receptor α (ERα) plays a role in controlling the expression and subcellular localization of ERα as well as its sensitivity to hormone response. Here, we show that ERα is also modified by UFM1 and this modification (ufmylation) plays a crucial role in promoting the stability and transactivity of ERα, which in turn promotes breast cancer development. The elevation of ufmylation via the knockdown of UFSP2 (the UFM1-deconjugating enzyme in humans) dramatically increases ERα stability by inhibiting ubiquitination. In contrast, ERα stability is decreased by the prevention of ufmylation via the silencing of UBA5 (the UFM1-activating E1 enzyme). Lys171 and Lys180 of ERα were identified as the major UFM1 acceptor sites, and the replacement of both Lys residues by Arg (2KR mutation) markedly reduced ERα stability. Moreover, the 2KR mutation abrogated the 17β-estradiol-induced transactivity of ERα and the expression of its downstream target genes, including pS2, cyclin D1, and c-Myc; this indicates that ERα ufmylation is required for its transactivation function. In addition, the 2KR mutation prevented anchorage-independent colony formation by MCF7 cells. Most notably, the expression of UFM1 and its conjugating machinery (i.e., UBA5, UFC1, UFL1, and UFBP1) were dramatically upregulated in ERα-positive breast cancer cell lines and tissues. Collectively, these findings implicate a critical role attributed to ERα ufmylation in breast cancer development by ameliorating its stability and transactivity.