• 혜화의학회지
• /
• 제13권2호
• /
• pp.131-146
• /
• 2004

In the present study, we exarnined anti-allergic effect of SGBHB in cultured B cells. B cells were prepared from isolated murine splenocytes and activated by co-treatment of anti-CD40 monoclonal antibody and recombinant IL-4 allergens. Anti-allergic effects of SGBHB in activated B cells were determined by measuring B cell surface activated molecules (CD23+ and CD11a+), and expression levels of IL-$1{\beta}$, IL-6, IL-10, TNF-$\alpha$, IgE, and HRF. The major findings are summarized as follows. 1. SGBHB treatment did not produce significant cytotoxic effects on mouse lung fibroblast cells. 2. SGBHB produced significant inhibitory effect on the expression of B cell surface activated molecules (CD23+ and CD11a) in activated B cells. 3. SGBHB treatment significantly inhibited expression levels of IL-$1{\beta}$, IL-6, and TNF-$\alpha$ mRNAs in activated B cells.IL-6 protein levels were significantly decreased by $100{\mu}g/m{\ell}$ of SGBHB treatrrient, and TNF-$\alpha$ protein levels were decreased compared to the control group, but statistically insignificant. 4. SGBHB treatment significantly increased IL-10 at both mRNA and protein levels in activated B cells. 5. SGBHB treatment significantly inhibited levels of IgE production. Thus, the present data suggest that SGBHB has an anti-allergic effect on activated B cells by controlling irnmune responses, and further implicates the possibility on clinical application as a therapeutic agent.

• 한국발생생물학회지:발생과생식
• /
• 제15권1호
• /
• pp.25-30
• /
• 2011

Embryonic stem (ES) cells can self-renew maintaining the undifferentiated state. Self renewal requires many factors such as Oct4, Sox2, FoxD3, and Nanog. NF-${\kappa}B$ is a transcription factor involved in many biological activities. Expression and activity of NF-${\kappa}B$ increase upon differentiation of ES cells. Reportedly, Nanog protein directly binds to NF-${\kappa}B$ protein and inhibits its activity in ES cells. Here, we found a potential binding site of NF-${\kappa}B$ in the human Nanog promoter and postulated that NF-${\kappa}B$ protein may regulate expression of the Nanog gene. We used human embryonic carcinoma (EC) cells as a model system of ES cells and confirmed decrease of Nanog and increase of NF-${\kappa}B$ upon differentiation induced by retinoic acid. Although deletion analysis on the DNA fragment including NF-${\kappa}B$ binding site suggested involvement of NF-${\kappa}B$ in the negative regulation of the promoter, site-directed mutation of NF-${\kappa}B$ binding site had no effect on the Nanog promoter activity. Furthermore, no direct association of NF-${\kappa}B$ with the Nanog promoter was detected during differentiation. Therefore, we conclude that NF-${\kappa}B$ protein may not be involved in transcriptional regulation of Nanog gene expression in EC cells and possibly in ES cells.

• BMB Reports
• /
• 제41권12호
• /
• pp.863-867
• /
• 2008

CD320 has been recently discovered and reported as a follicular dendritic cell (FDC) protein. Although CD320 is known to enhance proliferation of germinal center (GC) B cells, little other information is available. In this study, we investigated its cellular distribution in the GC. Confocal microscopy of human tonsil sections revealed co-localization of CD320 with CD19 and CD38 but not with CD3 indicating that GC B cells expressed CD320 in addition to FDC. In purified GC B cells, CD320 expression was inhibited in the nucleus, membrane and cytoplasm. Reverse transcriptase-polymerase chain reaction confirmed CD320 mRNA expression in B cells. These finding indicate that CD320 is expressed in B cells in addition to FDC, and that its GC activity may be more complicated than previously thought.

• 생명과학회지
• /
• 제26권12호
• /
• pp.1392-1399
• /
• 2016

Jaspine B는 석회해면류에서 추출된 sphingosine유도체로 인간 암세포에서의 항암활성이 보고되었다. 그러므로 본 연구는 다양한 인간 암세포주에서 항암활성을 비교하고, 암세포주에서의 Jaspine B의 농도를 측정하여 항암 활성과의 연관성을 확인하고자 하였다. 항암활성은 MTT 방법을 이용하여 측정하였고, $EC_{50}$ 값으로 표현하였다. 암세포주내 Jaspine B의 농도는 LC-MS/MS를 이용하여 분석하였다. 항암활성은 세포주마다 다양하게 나타났는데, 유방암과 흑색종 세포주에서 항암활성이 높게 나타났으며($EC_{50}$ 각각 $2.3{\mu}M$과 $2.6{\mu}M$), 신장암세포주에서는 $EC_{50}$ 값이 $29.4{\mu}M$이었다. 암세포주에서의 $EC_{50}$ 값은 동일한 세포에서의 Jaspine B 농도와 높은 상관성을 나타내었으며(r=0.838), 암세포내 약물농도를 조절하는 것으로 잘 알려진 P-glycoprotein과 breast cancer resistance protein 등의 배출수송계와는 관련이 없음을 확인하였다. 이상의 결과는 세포내 약물농도를 높게 유지하는 것이 항암활성에 매우 중요하며, 세포내 약물농도가 암세포주에 따라 다른 약효를 보이는 원인으로 사료된다.

• Toxicological Research
• /
• 제11권1호
• /
• pp.1-8
• /
• 1995

Fumonisins are a family of mycotoxins produced by the fungus Fusarium moniliforme which is a common contaminant in corn. Fumonisins are potent inhibitors of sphingosine and sphinganine N-acyltransferase (ceramide synthase), key enzymes in sphingolipid metabolism. The purpose of this study was to provide the evidence that the elevated levels of free sphingoid bases (primarily sphinganine) and depletion of complex sphingolipids were closely related to the inhibition of cell growth in LLC-$PK_1$ cells exposed to fumonisin $B_1$$(\leq 35 {\mu}M)$. Concentrations of fumonisin $B_1$ between 10 and $35 {\mu}M$ were known to inhibit cell growth without cytotoxicity in $LLC-PK_1$ cells (Yoo et al. Toxicol. Appl. Pharmacol. 114, 9-15, 1992). Cells exposed to 35$\mu M$ fumonisin B$_1$ for 48 and 72 hr developed a fibroblast-like (elongated and spindle-shaped) appearance and were less confluent than normal cells. At between 24 and 48 hr after exposure to fumonisin $B_1$ cells were beginning to show the inhibition of cell growth and at 72 hr the number of viable cells in fumonisin-treated cultures was about 50% of concurrent control cultures. During the 24 hr lag period preceding inhibition of cell growth, the free sphinganine levels in cells exposed to $35 {\mu}M$ fumonisin $B_1$ were highly elevated (approximately 230 fold higher than normal cells). The elevated levels of free sphinganine were $435\pm14$$pmoles/{10^6}$ cells at 48 hr and approximately TEX>$333\pm11$$pmoles/{10^6}$ cells in cells exposed to $35{\mu}M$ fumonisin$B_1$ at 72 hr, while the levels of free sphinganine in normal cells were less than 2$pmoles/{10^6}$ cells. Under the same condition, depletion of intracellular complex sphingolipids as a consequence of fumonisin inhibition of de novo sphingolipid biosynthesis and turnover pathway was appeared. Content of free sphingold bases in dividing cells was more elevated than in confluent cells at 24-48 hr after cells were exposed to $20{\mu}M$ fumonisin $B_1$. The dividing cells were showing the inhibition of cell growth at 48-72 hr and $20{\mu}M$ fumonisin $B_1$. The results of this study support the hypothesis that the inhibition of cell growth is very well related to the disruption of sphingolipid metabolism in $LLC-PK_1$ cells.

• BMB Reports
• /
• 제28권4호
• /
• pp.281-289
• /
• 1995

Homogeneous human deoxycytidine kinase was purified in one step from a variety of spontaneous human leukemic cells (T-ALL, B-ALL, B-CLL, AML, CML), and from cultured T-lymphoblast cells (MOLT-4) using the newly developed affinity medium, $dCp_4$-Sepharose. Starting with an ammonium sulfate fraction, purification was achieved in one step with the kinase being eluted from a column by the end product inhibitor, dCTP. The purified deoxycytidine kinase from T-ALL cells phosphorylated deoxyadenosine and deoxyguanosine, as well as deoxycytidine. The enzyme purified from T-ALL and B-CLL cells yielded one major band with a molecular weight of 52 kDa determined by SDS-polyacrylamide gel electrophoresis. AML and CML cells yielded one 52 kDa band and an extra band of 30 kDa molecular weight. On the other hand, B-ALL and MOLT-4 cells showed a low molecular weight band of 30 kDa only. However, the electrophoretic mobilities of enzymatic activity in 12% non-denaturing gels were identical for the dCyd kinase from all different kinds of leukemic cell lines, except that the B-ALL, B-CLL, and MOLT-4 cell preparations had an extra minor peak, all at the same position. dAdo and dCyd phosphorylating activities comigrated indicating that these activities are all associated with the same protein. Two new methods, a disk implantation method and a nitrocellulose powder method were used with a small amount of enzyme protein to raise polyclonal antibodies against dCyd kinase purified from T-ALL cells.

• 약학회지
• /
• 제46권1호
• /
• pp.75-80
• /
• 2002

Quercetin is one of the bioflavonoid compounds and has multiple biological effects such as antioxidant and effective anti-inflammatory agent. Melanin has an important role in protecting human skin from the damaging effects of ultra-violet W) radiation. We studied the effect of quercetin on proliferation of B16/F10 melanoma cells. After 48h treatment of cells with quercetin, the cells exhibited a dose-dependent inhibition in their proliferation without apoptosis. Therefore, the decrease in cell numbers may be due to cell growth arrest, not due to cell death by cytotoxicity. We also investigated the effect of quercetin on melanogenesis of this cells. B16/F10 melanoma cells were grown for 48h in the presence of 0.01~50$\mu\textrm{g}$/ml quercetin and the total melanin contents were measured. Quercetin stimulated melanization of the cells in low concentrations (0.01~20$\mu\textrm{g}$/ml), whereas it inhibited melanization in high concentrations (30~50$\mu\textrm{g}$/ml). It was observed that quercetin differently regulates melanogenesis of B16/F10 melanoma cells dependent on its concentrations.

• BMB Reports
• /
• 제50권12호
• /
• pp.640-646
• /
• 2017

Regulatory B cells, also well-known as IL-10-producing B cells, play a role in the suppression of inflammatory responses. However, the epigenetic modulation of regulatory B cells is largely unknown. Recent studies showed that the bromodomain and extra-terminal domain (BET) protein inhibitor JQ1 controls the expression of various genes involving cell proliferation and cell cycle. However, the role of BET proteins on development of regulatory B cells is not reported. In this study, JQ1 potently suppressed IL-10 expression and secretion in murine splenic and peritoneal B cells. While bromodomain-containing protein 4 (BRD4) was associated with $NF-{\kappa}B$ on IL-10 promoter region by LPS stimulation, JQ1 interfered the interaction of BRD4 with $NF-{\kappa}B$ on IL-10 promoter. In summary, BRD4 is essential for toll like receptor 4 (TLR4)-mediated IL-10 expression, suggesting JQ1 could be a potential candidate in regulating IL-10-producing regulatory B cells in cancer.

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

• IMMUNE NETWORK
• /
• 제20권6호
• /
• pp.50.1-50.11
• /
• 2020

Osteoporosis is prevalent in elderly women and it may cause dental implant failure. In particular, estrogen deficiency in postmenopausal women leads to higher rates of osteoporosis prevalence. Immune cell-mediated effects involving the development of osteoporosis have been studied previously; however, the role of IL-10-producing regulatory B (B10) cells in osteoporosis is largely unclear. Here, we examined the role of B10 cells in osteoporosis. C57BL/6 mice were subjected to ovariectomy (OVX). Fifteen weeks after OVX surgery, the first molar of the right maxillary was extracted, and twenty-four weeks after OVX surgery, serous progression of osteoporosis was observed in the alveolar bone. Moreover, the proportion of CD19⁺CD5⁺CD1d^high regulatory B cells, B10, and CD4⁺CD25⁺FoxP3⁺ regulatory T cells from the spleen of OVX mice decreased during the progression of osteoporosis, compared to controls. In contrast to regulatory cells, IL-17-producing Th (Th17) cell levels were increased in OVX mice. Adoptive transfer of B10 cells to OVX mice led to a decrease in Th17 cell abundance and inhibited the development of osteoporosis in the alveolar bone from OVX mice. Thus, our results suggest that B10 cells may help suppress osteoporosis development.