• 동의생리병리학회지
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• 제17권2호
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• pp.293-296
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• 2003

Prunellae Spica is a flower petal of Prunella vulgaris var. lilacina used for treatment of lymphoma, breast cancer, hepatitis and pathological fluid related diseases in oriental medicine. We tried to evaluate the mechanism of Prunellae Spica in the treatment of cancer. The ethyl acetate layer of Prunellae Spica showed a good cytotoxicity on U937 cells with IC50 of 8 ug/ml. It induced apoptosis in U937 dose-dependently by cell cycle analysis following PI staining. We also confirmed it induced DNA fragmentation in U937 cells from the concentration of 10 ug/ml. From western blot assay we observed the ethyl acetate layer of Prunellae Spica downregulated procaspase-3 and cleaved PARP in a dose dependent manner, whereas it didn't affect bax and bcl-2. Taken together, these results indicate the ethyl acetate layer of Prunellae Spica can induce apoptosis in U937 cells suggesting it can be potently applied to cancer.

• 대한한방부인과학회지
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• 제19권1호
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• pp.1-13
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• 2006

Purpose : Polygonum cuspidatum extract is an oriental herb which has been used for uterine diseases. In this study, the effects of Polygonum cuspidatum extract were investigated on inducing growth inhibition and apoptosis of human uterine cervical carcinoma cells. Methods : Viability of Polygonum cuspidatum extract-induced ME-180 cells was measured by MTT assay. Apoptotic cells were visualized by EtBr/AcOr staining under fluorescent microscope. Nucleosomal DNA fragmentation was assayed by agarose gel electrophoresis. Cell cycle distribution and changes in mitochondrial membrane potential were observed by flow cytometry. Results : Polygonum cuspidatum extract induced ME-180 cell death in a dose- and time-dependent manner. In the cells treated with Pc, the population of cells at sub-G1 phase significantly increased, and the condensed nuclei, apoptotic bodies and nucleosome-sized DNA were detected. Moreover, reduction in mitochondrial membrane potential was detected. Conclusion : Polygonum cuspidatum extract inhibits the growth and proliferation of ME-180 cells by apoptotic induction and facilitates its activity initiated by depolarization of mitochondria.

• 대한한의학방제학회지
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• 제22권2호
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• pp.35-43
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• 2014

Objectives : The purpose of this study was to identify antiproliferative effects of Salviae miltiorrhizae Radix(SM) extracts against cancer cell lines. Methods : We used 2 kinds of cancer cell lines such as colon cancer cells(HT-29), human oral epitheloid carcinoma cells(KB). MTT assay was performed to examine the efficacy of SM extracts on the cytostaticity of cancer cells in proportion to time and doses. Apoptosis was evaluated by DNA laddering and DAPI nuclei staining. Results : The MTT absorbances against HT-29 and KB of SM extracts were significantly decresed. DNA ladders could be identified in KB of SM extracts. The morphological change were observed and number of cells were decreased by SM extracts. Conclusions : SM extracts is considered to be effective to induce apoptosis and inhibit cancer cell proliferation.

• Natural Product Sciences
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• 제16권2호
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• pp.88-92
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• 2010

We verified that the apoptosis activities were examined by DNA fragmentation, flow cytometric analysis with annexin V staining, activation of caspase-3, and cytochrome c release. In the result, $\alpha$- and $\beta$-eudesmol induced DNA fragmentation in HL-60 cells at a concentration of $80\;{\mu}M$, respectively. Additionally, pro-apoptotic cells sorted by flow cytometry analysis were detected in HL-60 cells to 31.77 and 29.67% with $\acute{a}$- and $\beta$-eudesmol of $80\;{\mu}M$. Thus, both $\alpha$- and $\beta$-eudesmol exerted caspase-3 activation and cytochrome c release at $80\;{\mu}M$ in HL-60 cells. These results are firstly reported that the sesquiterpenes, $\alpha$- and $\beta$-eudesmol are apoptosis inducers through mitochondria-dependent caspase cascade in HL-60 cells.

• 생명과학회지
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• 제17권3호통권83호
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• pp.420-426
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• 2007

본 연구에서는 유전자 발현에 중요한 조절 역할을 하는 DNA 메틸화를 식이성 폴리페놀의 하나인 녹차의 대표적인 추출물 EGCG로 억제하였을 때 C2C12 myoblast 세포에 일어나는 현상을 살펴보았다. 그 결과 smooth muscle의 지표인 transgelin, smooth muscle ${\alpha}-actin$ mRNA와 단백질이 현저히 증가함을 보였고, 형태학적으로도 smooth muscle의 전형적인 모습을 보였다. 식이에 포함된 DNA 메틸화 억제제인 EGCG가 C2C12 myoblast를 smooth muscle로 분화를 유도하였으며, 암 예방 차원에서의 EGCG의 역할 외에 혈관질환과 같은 smooth muscle에 관련된 예방과 치료차원에서 EGCG의 역할이 있을 것으로 사료된다. 본 연구는 C2C12 myoblast를 smooth muscle로 유도하는 결정적인 신호전달 역할을 하는 유전자에 대한 연구는 수행하지 못하였다. 따라서 EGCG에 의해 변화되는 유전자에 대한 기전연구가 필요하다고 하겠다.

• Journal of Oral Medicine and Pain
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• 제34권3호
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• pp.261-276
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• 2009

Sreptomyces라는 세균에서 추출한 lactacystin은 선택적인 proteasome 억제제로서 많은 연구에서 사용되어져 왔다. Proteasome 억제제는 최근의 많은 연구를 통해서 암세포증식의 억제에 대한 효과가 증명되었으며, 특히 다른 항암제와 병용처리 시, 상호작용에 의한 상승효과가 있다고 알려져 있다. 현재 proteasome 억제제는 새로운 강력한 항암제로서 분류되어 있다. 본 연구는 사람혀편평세포암종세포(SCC25 cells)에서 lactacystin의 세포독성과 성장억제 효과, 그리고 세포자멸사의 유도에 대한 분자생물학적 기전을 밝히기 위해 실험을 시행하였다. SCC25 세포, 사람정상각화세포 (HaCaT cells) 그리고 사람치은섬유모세포(HGF-1 cells)의 생존율 측정은 MTT법을 시행하였고, SCC25 세포의 성장억제를 확인하기 위해서는 clonogenic assay를 사용하였다. lactcystin이 SCC25 세포에서 세포자멸사가 유도되는지를 확인하기 위해서 hoechst 염색법, hemacolor 염색법 그리고 TUNEL법을 시행하였다. 그리고 SCC25 세포에 lactacystin을 적용한 후, Western blot 분석, 세포면역화학염색, 공초점레이저주사현미경 검경, FACScan flow cytometry, 사립체막 전위변화, proteasome 활성도 측정 등을 시행하였다. Lactacystin으로 처리된 SCC25 세포는 시간 및 용량 의존적인 세포생존율의 감소, 용량의존적인 세포성장억제 그리고 세포자멸사에 의한 세포죽음을 보였다. 흥미롭게도 lactacytin은 정상세포인 HaCat 세포와 HGF-1 세포에서는 세포독성을 전혀 보이지 않았다. 그리고 lactacystin이 적용된 SCC25세포에서 핵 응축, DNA의 조각남, 사립체막전위와 proteasome 활성도의 감소, DNA 양의 감소, cytochrome c의 사립체에서의 세포질로의 유리, AIF와 DFF40 (CAD)의 핵으로의 이동, Bax의 증가, caspase-7, caspase-3, PARP, lamin A/C 그리고 DFF45 (ICAD)의 활성화 혹은 파괴와 같은 아주 다양한 세포자멸사 증거를 보였다. Flow cytometry 분석에서는 CDK 억제제인 $p21^{WAF1/CIP1}$와 $p27^{KIP1}$의 발현 증가와 관계있는 것으로 추정되어 지는 G1 세포주기 정지를 보였다. 이러한 결과는 lactacytin이 SCC25 세포에서 G1 세포주기정지와 proteasome, 사립체 및 caspase 경로의 연속반응을 통한 세포자멸사를 유도함을 명확하게 증명하고 있다. 이와 같은 세포주기 정지와 세포자멸사 유도능은 lactacytin이 사람혀편평상피세포암종의 새로운 치료전략으로서의 가능성을 제공한다고 생각한다.

• 한국산림과학회지
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• 제80권2호
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• pp.232-236
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• 1991

형광색소(螢光色素) berberine sulphate의 DNA 및 RNA-결합성질(結合性質)을 이용(利用)한 형광현미경기법(螢光顯微鏡技法)의 마이코플라스마(mycoplasma-like organism ; MLO) 감염진단(感染診斷)에 대(對)한 효용가치(效用價値)를 조사(調査)하였다. 이병(罹病) 대추나무, 오동나무, 뽕나무 및 일일초(日日草)의 조직절편(組織切片)을 berberine sulphate로 염색(染色)하여 형광현미경(螢光顯微鏡)으로 관찰(觀察)하였던 바, 사부조직(篩部組織)에서 MLO-특이형광반응(特異螢光反應)이 나타났으나, 건전조직(健全組織)의 사부(篩部)에서는 특이형광(特異螢光)이 관찰(觀察)되지 않았다. 이와 같은 berberine sulphate의 조직염색법(組織染色法)은 목본(木本) 및 초본식물(草本植物)의 MLO감염(感染)을 정확(正確)하게 진단(診斷)하는데 매우 유용(有用)한 방법(方法)임을 보여 주었으며, 사용(使用)이 간단(簡單)하고 비용(費用)이 저렴(低廉)하므로 대량(大量)의 시료(試料)에 대한 신속(迅速)한 검정(檢定)에 이용(利用)될 가능성(可能性)을 보여 주었다.

• Journal of Microbiology and Biotechnology
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• 제29권12호
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• pp.1969-1974
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• 2019

A Gram-staining-negative, aerobic, light brown pigment bacterium, designated strain CE80^T was isolated from marine sponge Callyspongia elegans in Jeju Island, Republic of Korea. Strain CE80^T grew optimally at 25℃, in the range of pH 5.0-11.0 (optimum 7.0-8.0), and with 1.0-5.0% NaCl (optimum 1-3% (w/v)). Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain CE80^T belonged to the genus Labrenzia and was closely related to L. suaedae YC6927^T (98.3%), L. alexandrii DFL-11^T (96.6%), L. aggregata IAM 12614^T (96.6%) L. marina mano18^T (96.5%) and L. alba CECT 5094^T (96.2%). The major fatty acids of strain CE80^T were C_18:1 ω7c, and summed feature. The polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylmonomethylethanolamin, one unidentified aminolipid, one phospholipid and four unidentified lipids. The DNA G+C content of strain CE80^T was 55.9 mol%. The major respiratory quinone was Q-10. DNA-DNA relatedness between strain CE80^T and L. suaedae YC6927^T was 56.1±2.8%. On the basis of physiological and biochemical characterization and phylogenetic and chemotaxonomic analysis, strain CE80^T represents a novel species of the Labrenzia, for which the name Labrenzia callyspongiae sp. nov., is proposed. The type strain is CE80^T (=KCTC 42849^T =JCM 31309^T).

• International Journal of Oral Biology
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• 제40권1호
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• pp.51-61
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• 2015

Shikonin, a major ingredient in the traditional Chinese herb Lithospermumerythrorhizon, exhibits multiple biological functions including antimicrobial, anti-inflammatory, and antitumor effects. It has recently been reported that shikonin displays antitumor properties in many cancers. This study was aimed to investigate whether shikonin could inhibit oral squamous carcinoma cell (OSCC) growth via mechanisms of apoptosis and cell cycle arrest. The effects of shikonin on the viability and growth of OSCC cell line, SCC25 cells were assessed by MTT assay and clonogenic assays, respectively. Hoechst staining and DNA electrophoresis indicated that the shikonin-treated SCC25 cells were undergoing apoptosis. Western blotting, immunocytochemistry, confocal microscopy, flow cytometry, MMP activity, and proteasome activity also supported the finding that shikonin induces apoptosis. Shikonin treatment of SCC25 cells resulted in a time- and dose-dependent decrease in cell viability, inhibition of cell growth, and increase in apoptotic cell death. The treated SCC25 cells showed several lines of apoptotic manifestation as follows: nuclear condensation; DNA fragmentation; reduced MMP and proteasome activity; decrease in DNA contents; release of cytochrome c into cytosol; translocation of AIF and DFF40 (CAD) onto the nuclei; a significant shift in Bax/Bcl-2 ratio; and activation of caspase-9, -7, -6, and -3, as well as PARP, lamin A/C, and DFF45 (ICAD). Shikonin treatment also resulted in down-regulation of the G1 cell cycle-related proteins and up-regulation of $p27^{KIP1}$. Taken together, our present findings demonstrate that shikonin strongly inhibits cell proliferation by modulating the expression of the G1 cell cycle-related proteins, and that it induces apoptosis via the proteasome, mitochondria, and caspase cascades in SCC25 cells.

• 대한수의학회지
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• 제39권1호
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• pp.148-158
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• 1999

The purpose of this study was to establish a rapid, reliable diagnostic method detecting Encephalomyocarditis virus(EMCV) RNA in formalin-fixed, paraffin-embedded tissues of EMCV naturally infected pigs by cDNA probe of EMC $K_3$, the EMCV strain isolated from Korea. Using a biotin-labelled nick translated probe for the cDNA marker. We made up for some defects of radiolabeled method. In sits hybridization(ISH) technique, differently from the other nucleic acid hybridization methods, is able to detect the virus genome specifically in the state of the intact shapes of cells and/or tissues. We succeeded in performing the experiment to detect the EMCV within 1~2 hours using the $MicroProbe^{TM}$ capaillary action system. In this study, we observed highly specific positive signals of red color by staining the paraffin-embedded tissue sections of naturally EMCV-infected pig organs or tissues, including brain, heart, kidney and lacrimal gland with the Fast Red TR salt/Naphtol phosphate chromogen. The results suggested that this ISH method is considered as a highly sensitive and reliable tool for molecular biologic diagnosis of the EMC viral disease.