• BMB Reports
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• 제30권4호
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• pp.269-273
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• 1997

Heptocvte-specific expression induced by Hepatitis B virus (HBV) enhancer 2-core gene promoter was examined in various hepatocyte and non-hepatocyte cell lines. using non-viral and retroviral vector systems in which chloramphenicol acetyltransferase (CAT) is used as a reporter. The non-viral plasmid containing the HBV enhancer 2-core promoter exhibited 22 and 66% of CAT activities in hepatoma cell lines. HepG2 and Hep3B, respectively when compared with CAT activity expressed by CMV promoter. The CAT activities, however. were found to be marginal in other tested hepatoma cell lines as well as mouse primary hepatocytes and non-hepatocytes. The HBV enhancer 2 located upstream the CMV promoter did not affect the CMV promoter activity nor provided hepatocyte-specific expression. Transfection of retroviral plasmid DNA containing the HBV enhancer 2-core promoter as an internal promoter exhibited high and specific CAT expression in HepG2 and Hep3B cell lines but the activity value was 5 to 10 fold lower than the non-viral plasmid with identical promoter. These results suggest that the usage of HBV enhancer 2-core promoter for liver specific expression is limited to certain vectors and hepatocyte cell lines.

• 한방안이비인후피부과학회지
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• 제22권1호
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• pp.108-119
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• 2009

Objectives : This study was carried out to investigate the effects of Samhwangseje(SHSJ) on anti-Inflammation in Raw 264.7 cell. Methods : The effects of SHSJ on anti-Inflammation were measured by the cytotoxicity of Raw 264.7 cell, the inhibition for NO, TNF-$\alpha$, $PGE_{2}$, iNOS and COX-2, the blocking NF-${\kappa}B$ into nucleus. Results : 1. All concentrations of SHSJ had no cytotoxicity in Raw 264.7 cell. 2. All concentrations of SHSJ inhibited the production of NO in the Raw 264.7 cell stimulated with LPS. 3. All concentrations of SHSJ did not inhibit the production of TNF-$\alpha$ in the Raw 264.7 cell stimulated with LPS. 4. All concentrations of SHSJ inhibited the production of $PGE_{2}$ in the Raw 264.7 cell stimulated with LPS. 5. All concentrations of SHSJ did not inhibit the expression of COX-2 but concentrations of 50 ${\mu}g/ml$, 100 ${\mu}g/ml$ SHSJ inhibited iNOS expression in the Raw 264.7 cell stimulated with LPS. 6. Concentrations of 50 ${\mu}g/ml$, 100 ${\mu}g/ml$ SHSJ had the effect of blocking NF-${\kappa}B$ into nucleus in LPS-induced macrophage Raw 264.7 cell.

• Toxicological Research
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• 제22권1호
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• pp.31-37
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• 2006

It was previously found that melittin inhibited $NF-{\kappa}B$ activity by reacting with signal molecules of $NF-{\kappa}B$ which is critical contributor in cancer cell growth by induction of apoptotic cell death. We here investigated whether melittin inhibits cell growth of human prostate cancer cells through induction of apoptotic cell death, and the possible signal pathways. Melittin ($0{\sim}1\;{\mu}g/ml$) inhibited prostate cancer cell growth in a dose dependent manner. Conversely related to the growth inhibitory effect, melittin increased the induction of apoptotic cell death in a dose dependent manner. Melittin also inhibited DNA binding activity of $NF-{\kappa}B$, an anti-apoptotic transcriptional factor. Consistent with the induction of apoptotic cell death and inhibition of $NF-{\kappa}B$, melittin increased the expression of pro-apoptotic proteins caspase-3, and Bax but down-regulated anti-apoptotic protein Bcl-2. These findings suggest that melittin could inhibit prostate cancer cell growth, and this effect may be related with the induction of apoptotic cell death via inactivation of $NF-{\kappa}B$.

• Asian Pacific Journal of Cancer Prevention
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• 제13권3호
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• pp.931-935
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• 2012

Cdc25 phosphatases are important regulators of the cell cycle. Their abnormal expression detected in a number of tumors implies that their dysregulation is involved in malignant transformation. However, the role of Cdc25B in renal cell carcinomas remains unknown. To shed light on influence on renal cell carcinogenesis and subsequent progression, Cdc25B expression was examined by real-time RT-PCR and western blotting in renal cell carcinoma and normal tissues. 65 kDa Cdc25B expression was higher in carcinomas than in the adjacent normal tissues (P<0.05), positive correlations being noted with clinical stage and histopathologic grade (P<0.05). To additionally investigate the role of Cdc25B alteration in the development of renal cell carcinoma, Cdc25B siRNA was used to knockdown the expression of Cdc25B. Down-regulation resulted in slower growth, more G2/M cells, weaker capacity for migration and invasion, and induction of apoptosis in 769-P transfectants. Reduction of 14-3-3 protein expression appeared related to Cdc25B knockdown. These findings suggest an important role of Cdc25B in renal cell carcinoma development and provide a rationale for investigation of Cdc2B-based gene therapy.

• Biomolecules & Therapeutics
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• 제11권2호
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• pp.151-156
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• 2003

Cornis fructus have various biological effects and major chemical components have been tannins, saponins, ursolic acids, gallic acids, linoleic acids, morronisides, cornins and loganins. The main aim of the present study is measurment the effect of chloroform extract from Cornis fructus on proliferation inhibition and Cell death. Cells were cultured in the presence of chloroform extracts from Cornis fructus for 48 h. after 48h treatment of B16/F10 melanoma cells with chloroform extracts, the cells were observed a dose-dependent inhibitions of cell viability with cell death in their proliferation. the cells were estimated cell viability, cell number, total DNA fragmentation and chromatin condensation in a dose-dependent manner. It also caused cell death as measured by cell morphology, DNA fragmentation and nucleus chromatin condensation. therefore, these results suggest that chloroform extracts from C. fructus is inhibitory proliferation and is related to cell death in this cells.

• 대한임상검사과학회지
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• 제52권3호
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• pp.245-252
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• 2020

대식세포사멸은 죽상판 형성에 영향을 미쳐 죽상동맥경화증 발병에 관여하는 것으로 알려져 있다. 중성지방 역시 죽상동맥경화 발병에 기여한다고 알려져 있는데 최근 본 연구팀에서는 중성지방이 대식세포사멸을 유발한다는 결과를 확인하였다. 본 연구에서는 cathepsin B가 중성지방에 의해 유발되는 대식세포사멸 과정에 관여하는지 확인하고자 연구를 진행하였다. THP-1 대식세포에 중성지방 처리 시 cathepsin B의 발현량에는 변화가 없고 리소좀에 있던 cathepsin B가 세포질로 방출되어 세포질의 cathepsin B가 증가한 것을 확인하였다. 다음으로 cathepsin B 억제제인 CA-074 Me를 처리 시 중성지방에 의해 유도되는 대식세포사멸이 일부 회복되는 것을 확인하였다. 본 연구팀의 이전 연구에서 중성지방에 의한 대식세포사멸이 caspase-1, -2 및 apoptotic caspase 활성화를 매개로 일어남을 확인하였기 때문에 본 연구에서는 이러한 caspase 활성 경로와 cathepsin B와의 연관성에 대해 연구하였다. cathepsin B 억제시 caspase-7, -8 및 -1의 활성은 억제되었으나, caspase-3, -9 및 -2는 활성에는 영향을 미치지 않음을 알 수 있었다. 정리하면, 중성지방에 의해 세포질로 방출된 cathepsin B는 caspase-1 활성화에 기여하고, 활성화된 caspase-1은 외인성 apoptotic caspase 경로를 활성화하여 THP-1 대식세포 사멸을 유발한다는 것을 알 수 있다.

• 한국식품영양과학회지
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• 제32권3호
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• pp.428-436
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• 2003

Carotenoids는 항암 효과가 있는 것으로 알려져 있으나 각각의 carotenoids가 대장암에 미치는 영향에 대해서는 명확하게 밝혀진 바가 없다. 본 연구에서는 4가지 종류의 carotenoids가 인간의 대장에서 유래한 암세포인 HT-29 세포의 증식에 미치는 영향을 조사하였다. $\alpha$-carotene, $\beta$-carotene, lutein lycopene을 농도를 달리 하여 세포 배양액 에 첨가하여 살아있는 세포의 수를 측정한 결과 $\beta$-carotene는 세포의 증식을 다소 증가시키는 반면 $\alpha$-carotene, lutein, lcopene은 세포의 증식을 감소하였다. 세포의 증식을 억제한 carotenoids 중에서 lycopene이 그 효과가 가장 컸다. ErbB receptor family는 세포의 증식을 촉진하고 대장암에서 그 발현이 증가된 것으로 보고되었기 때문에 lycopene이 heregulin-ErbB3 signaling을 억제하는지를 조사하였다. Lycopene는 ErbB2 단백질을 감소하였고 ErbB3 단백질의 변화를 초래하였다. Heregulin을 첨가하여 인산화를 유도한 경우 ErbB3의 인산화, ErbB3와 p85의 결합, Akt 인산화가 lycopene에 의해 억제되었다. 이 결과들은 carotenoids 중 lycopene이 대장암 세포 증식 억제 효과가 가장 크고, 대장암 세포의 DNA 합성을 억제하고 apoptosis를 유도하는 lycopene효과의 일부는 Erb-B3와 Akt의 인산화 감소에 기인하는 것임을 나타낸다.

• Asian Pacific Journal of Cancer Prevention
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• 제17권2호
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• pp.491-495
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• 2016

Background: Small B-cell non-Hodgkins lymphoma (NHL) is difficult to be distinguished from non-neoplastic reactive processes using conventional haematoxylin-eosin (HE) staining due to different interpretations among pathologists with diagnosis based on morphologic features. Ancillary examinations such as immunohistochemical (IHC) staining are essential. However, negative or doubtful results are still sometimes obtained due to unsatisfactory tissue processing or IHC technique. The polymerase chain reaction (PCR) as a molecular diagnostic technique is very sensitive and specific. Clonality detection of heavy chain immunoglobulin (IgH) gene rearrangement has been widely used to establish diagnosis of B-cell NHL. Aims: To elaborate interobserver variation in small B-cell NHL diagnosis based on morphologic features only and to confirm sensitivity and specificity of the PCR technique as an ancillary method. Materials and Methods: A toptal of 28 samples of small B cell NHL and suspicious lymphoma were interpreted by 3 pathologists in Sardjito General Hospital based on their morphology only. The reliability of assessment and the coefficient of interobserver agreement were calculated by Fleiss kappa statistics. Interpretation results were confirmed with IHC staining (CD20, CD3, Bcl2). PCR was performed to analyze the clonality of IgH gene rearrangement. Results: Interobserver agreement in morphologic evalution of small B cell NHL and chronic lymphadenitis revealed kappa coefficient 0.69 included in the substantial agreement category. The cases were divided into 3 groups based on morphology and IHC results; lymphoma, reactive process and undetermined group. PCR analysis showed 90% sensitivity and 60% specificity. Conclusions: The present study revealed a substantial agreement among pathologists in small B-cell NHL diagnosis. For difficult cases, PCR is useful as complementary method to morphologic and IHC examinations to establish definitive diagnosis.

• 치위생과학회지
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• 제23권3호
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• pp.216-224
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• 2023

Background: Smilax glabra has various pharmacological activities and is widely used as a herbal medicine. Although the incidence of oral cancer is low, the recurrence rate is high, and the 5-year survival rate is poor. It is necessary to search for anticancer drugs that increase the effect of cancer chemotherapy on heterogeneous oral tissues and reduce the side effects on normal cells. This study aimed to investigate the effects and mechanism of ethanol extract of Smilax glabra (EESG) as an anticancer drug for oral cancer. Methods: Smilax glabra root components extracted with 70% ethanol were used to analyze their effects on cancer cells. A 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide assay was performed for cytotoxicity analysis. Flow cytometry was performed to determine the cell cycle phase distribution. To observe apoptotic cells, terminal deoxynucleotidyl transferase dUTP nick end labeling and γH2AX were detected by fluorescence microscope. The protein levels of cleaved PARP and caspase were analyzed using western blotting. The activation of procaspase-3 was confirmed by measuring caspase-3 activity. Results: EESG was no cytotoxic to normal gingival fibroblast but was high in YD10B oral squamous cell carcinoma (OSCC) cells. EESG treatment increased the subdiploid DNA content of YD10B cells by assessing DNA content distribution. Chromatin condensation and DNA strand breaks increased in YD10B cells treated with EESG. EESG-treated YD10B cells had high Annexin V and low propidium iodide levels, confirming that early apoptosis was induced. In addition, increased levels of γH2AX foci, a marker of DNA damage, were observed in the nuclei of EESG-treated YD10B cells. The EESG-treated YD10B cells also exhibited decreased procaspase-3 and procaspase-9 levels, increased PARP cleavage and caspase-3 activity. Conclusion: These results indicate that EESG inhibited cancer cell proliferation by inducing apoptosis in YD10B OSCC cells.

• Molecules and Cells
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• 제40권10호
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• pp.752-760
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• 2017

A2B adenosine receptor (A2BAR) is known to be the regulator of bone homeostasis, but its regulatory mechanisms in osteoclast formation are less well-defined. Here, we demonstrate the effect of A2BAR stimulation on osteoclast differentiation and activity by RANKL. A2BAR was expressed in bone marrow-derived monocyte/macrophage (BMM) and RANKL increased A2BAR expression during osteoclastogenesis. A2BAR stimulation with its specific agonist BAY 60-6583 was sufficient to inhibit the activation of ERK1/2, p38 MAP kinases and $NF-{\kappa}B$ by RANKL as well as it abrogated cell-cell fusion in the late stage of osteoclast differentiation. Stimulation of A2BAR suppressed the expression of osteoclast marker genes, such as c-Fos, TRAP, Cathepsin-K and NFATc1, induced by RANKL, and transcriptional activity of NFATc1 was also inhibited by stimulation of A2BAR. A2BAR stimulation caused a notable reduction in the expression of Atp6v0d2 and DC-STAMP related to cell-cell fusion of osteoclasts. Especially, a decrease in bone resorption activity through suppression of actin ring formation by A2BAR stimulation was observed. Taken together, these results suggest that A2BAR stimulation inhibits the activation of ERK1/2, p38 and $NF-{\kappa}B$ by RANKL, which suppresses the induction of osteoclast marker genes, thus contributing to the decrease in osteoclast cell-cell fusion and bone resorption activity.