• BMB Reports
• /
• v.57 no.5
• /
• pp.244-249
• /
• 2024

Many types of cancer are associated with excessive angiogenesis. Anti-angiogenic treatment is an effective strategy for treating solid cancers. This study aimed to demonstrate the inhibitory effects of (E)-2-methoxy-4-(3-(4-methoxyphenyl) prop-1-en-1-yl) phenol (MMPP) in VEGFA-induced angiogenesis. The results indicated that MMPP effectively suppressed various angiogenic processes, such as cell migration, invasion, tube formation, and sprouting of new vessels in human umbilical vein endothelial cells (HUVECs) and mouse aortic ring. The inhibitory mechanism of MMPP on angiogenesis involves targeting VEGFR2. MMPP showed high binding affinity for the VEGFR2 ATP-binding domain. Additionally, MMPP improved VEGFR2 thermal stability and inhibited VEGFR2 kinase activity, suppressing the downstream VEGFR2/AKT/ERK pathway. MMPP attenuated the activation and nuclear translocation of NF-κB, and it downregulated NF-κB target genes such as VEGFA, VEGFR2, MMP2, and MMP9. Furthermore, conditioned medium from MMPP-treated breast cancer cells effectively inhibited angiogenesis in endothelial cells. These results suggested that MMPP had great promise as a novel VEGFR2 inhibitor with potent anti-angiogenic properties for cancer treatment via VEGFR2/AKT/ERK/NF-κB signaling pathway.

• Tuberculosis and Respiratory Diseases
• /
• v.41 no.3
• /
• pp.215-221
• /
• 1994

Background: It is well known that oxygen free radicals(OFR) play a vital role in the various type of acute lung injury. Among various antioxidant defense mechanisms, the superoxide dismutases(SOD) are thought to be the first line of antioxidant defense by catalyzing the dismutation of two superoxide radicals to yield hydrogen peroxide and oxygen. Eukaryotic cells contain two types of intracellular SOD : cytosolic, dimeric copper/zinc- containing enzyme(CuZnSOD) and mitochondrial, tetrameric manganese-containing enzyme(MnSOD). The purpose of this study is to evaluate the time-dependent gene expression of MnSOD and CuZnSOD in the endotoxin-treated rats, and to compare with the manifestations of LPS-induced acute lung injury in rats. Methods: Total RNA from rat lung was isolated using single step phenol extraction 0, 1, 2, 4, 6, 12, 18, 24 hours after E. coli endotoxin injection(n=3, respectively). RNA was separated by formaldehyde-containing 1.2% agarose gels elctrophoresis, transblotted, baked, prehybridized, and hybridized with $^{32}P$-labeled cDNA probes for rat MnSOD and CuZnSOD, which were kindly donated by Dr. Ho(Duke University, Durham, NC, USA). The probes were labeled by nick translation. Blots were washed and autoradiography were quantitated using laser densitometry. Equivalent amounts of total RNA/gel were assessed by monitoring 28S and 18S rRNA. Results: Endotoxin caused a rise in steady-state MnSOD mRNA levels by 4h with peak mRNA accumulation by 6h. Continued MnSOD mRNA expression was observed at 12h. CuZnSOD mRNA expression was observed from 1h to 24h with peak levels by 18h. Conclusion: These results suggest that SOD palys an important defensive role in the endotoxin-induced acute lung injury in rats.

• Imaging Science in Dentistry
• /
• v.35 no.3
• /
• pp.127-131
• /
• 2005

Purpose : We performed the present study to investigate whether osteotropic hormomes play roles on the nitric oxide (NO) production in culture of ROS 17/12.8 osteoblastic cells. Materials and Methods : The osteoblastic cell line ROS17/2.8 cells were cultured In F12 medium supplemented with $5\%$ fetal bovine serum (FBS) at $37^{\circ}C$ in a humidified atmosphere of $5\%\;CO_2$ in air. ROS17/2.8 cells were plated in 96-well plates at a density of $2-3\times10^3cells/well$ and grown to confluence. Then the cells were pretreated with osteotropic hormones (parathyroid hormone (PTH) 20-500 ng/mL, 1, 25-dihydroxycholecalciferol $(1,\;25[OH]_2D_3)$ 1-100 nM; prostaglandin $E_2 (PGE_2)$ 20-500 ng/mL in the medium supplemented with $0.4\%$ FBS for 72 hours and the cells were treated with cytokines $(TNF{\alpha}\;and\;IFN{\gamma})$ in phenol red-free F12 medium for an additional 48 hours. NO synthesis was assessed by measuring the nitrite anion concentration, the reaction product of NO, in the cell culture medium using Griess reagent. Results : PTH and $1,\;25[OH]_2D_3$ pretreatment induced a significant increase in NO production in the presence of $TNF{\alpha}\;and\;IFN{\gamma}.\;PGE_2$ slightly induced NO production compared to the control group. But, $PGE_2$ pretreatment did not affect in NO production in the presence of $TNF{\alpha}\;and\;IFN{\gamma}$. Conclusions : These results suggest that the actions of osteotropic hormones In bone metabolism may be partially mediated by NO in the presence of cytokines.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.26 no.6
• /
• pp.581-590
• /
• 2000

Alterations in the cellular genome affecting the expression or function of genes controlling cell growth and differentiation are considered to be the main cause of cancer. Over 30 oncogenes can be activated by insertional mutagenesis, single point mutations, chromosomal translocations and gene amplification. The ras oncogenes have been detected in $15{\sim}20%$ of human tumors that include some of the most common forms of human neoplasia and are known to acquire their transforming properties by single point mutations in two domains of their coding sequences, most commonly in codons 12 and 61. The ras gene family consists of three functional genes, N-ras, K-ras and H-ras which encode highly similar proteins of 188 or 189 amino acid residues generically known as P21. ras proteins have been shown to bind GTP and GTP, and possess intrinsic GTPase activity. Experimental study was performed to observe the mutational change of the ras gene family and apply the results to the clinical activity. 36 Golden Syrian Hamster each weighing $60{\sim}80g$ were used and painted with 0.5% DMBA by 3 times weekly on the right buccal cheek(experimental side) for 6, 8, 10, 12, 14 and 16 weeks. Left buccal cheek (control side) was treated with mineral oil as the same manner of the right side. The hamsters were sacrificed on the 6, 8, 10, 12, 14 & 16 weeks. Normal and tumor tissues from paraffin block were completely dissected by microdissection and DNA from both tissue were isolated by proteinase K/phenol/chloroform extraction. Segments of the K-ras and H-ras gene were amplified by PCR using the oligonucleotide primers corresponding to the homologous region (codon 12 and 61) of the hamster gene, and then confirmational change of ras genes was observed by SSCP and autosequencing analysis. The results were as follows : 1. Malignant lesion could be found in the experimental side from the experimental six weeks. 2. One hamster among six showed point mutation of the H-ras codon 12($G{\rightarrow}A$ transition) at the experimental 10 and 14 weeks. 3. One of six at 6 weeks, two of six at 8 weeks and one of six at 12 weeks revealed the confirmational change of the H-ras codon 61($A{\rightarrow}T$ transversion). 4. The incidence of point mutation of H-ras codon 12 and 61 were 5.5%(2 of 36) and 11%(4 of 36) respectively. 5. Point mutation of the K-ras could not be seen during the whole experimental period. Form the above results, these findings strongly support the concept that H-ras oncogenes may have the influence of the DMBA induced carcinoma of hamster buccal pouch.

• Journal of Life Science
• /
• v.18 no.6
• /
• pp.814-825
• /
• 2008

Ascochlorin (ASC) is prenyl-phenol compound that was isolated from the fungus Ascochyta viciae. ASC reduces serum cholesterol and triglyceride levels, and suppresses hypertension, tumor development, ameliorates type I and II diabetes. Here, to better understand the mechanisms by which ASC regulates physiological or pathological events and induces responses in the pharmacological treatment of inflammation, we performed differential analysis of the proteome of the mouse macrophage RAW264.7 cells in response to ASC. In this study, we used a proteomic analysis of LPS-induced RAW264.7 cells treated by ASC, to identify proteins potentially involved in inflammatory processes. The RAW264.7 cell proteomes with and without treatment with ASC were compared using two-dimensional electrophoresis (2-D SDS-PAGE), matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF-MS) and bioinformatics. The largest differences in expression were observed for the calreticulin (4-fold decrease), ${\beta}-actin$ (4-fold decrease) and vimentin (1.5-fold decrease). In addition, rabaptin was increased 3-fold in RAW264.7 cells treated with ASC. The expression of some selected proteins was confirmed by RT-PCR analysis.

• The Korean Journal of Physiology and Pharmacology
• /
• v.2 no.4
• /
• pp.493-501
• /
• 1998

The most significant direct role of estrogen in vivo is its ability to elicit receptor-mediated cellular proliferation in mammalian target tissues. However, the mechanism by which exogenously added estrogen causes the neoplastic transformation of renal cortical cells is yet to be uncovered. The present study was designed to evaluate interaction of $17{\beta}-estradiol\;(E_2)$ with epidermal growth factor (EGF) and insulin-like growth factor-I (IGF-I) on proliferation and $P_i$ uptake in primary cultured rabbit renal proximal tubular cells in phenol red-free, hormonally defined-medium. $[^3H]-thymidine$ incorporation increased markedly by about 133% and 141% more in the presence of $10^{-9}\;and\;10^{-6}\;M\;E_2$, respectively, than that of control. Cell count was 162% and 143% greater in the presence of $10^{-9}\;and\;10^{-6}\;M\;E_2$ , respectively, compared with control. Among all time points examined, there was an increase in $[^3H]-thymidine$ incorporation in the presence of $10^{-9}\;M\;E_2$ at day 9 or 13, respectively. However, $E_2$ ($10^{-9}\;M$) significantly drove up cell count to 160% of that of control at day 13, while it had a slight but statistically insignificant effect at day 9. $E_2-induced$ stimulation of $[^3H]-thymidine$ incorporation was completely reversed by $E_2$ antagonists (progesterone or tamoxifen). $E_2$ ($10^{-9}\;M$) or EGF ($10^{-8}\;M$) significantly stimulated $[^3H]-thymidine$ incorporation by 144% and 154% of control. $E_2$ plus EGF was synergistic on $[^3H]-thymidine$ incorporation (204% of control), while $E_2$ plus IGF-I showed a slight but no significant synergistic effect. Cell number also displayed similar pattern. $E_2$ ($10^{-9}\;M$) significantly stimulated $P_i$ uptake to 134% of control. $E_2$-induced stimulation of $P_i$ uptake was partially reversed by $E_2$ antagonists. EGF or IGF-I ($10^{-8}\;M$) significantly also increased $P_i$ uptake to 132% or 129% of control. $E_2$ plus EGF had synergistic effect on $P_i$ uptake, while $E_2$ plus IGF-I did not. In conclusion, $E_2$ may act not only directly interaction with its receptors but also indirectly as a modulator of EGF in proliferation and $P_i$ uptake of primary cultured rabbit renal proximal tubular cells.

• Journal of Life Science
• /
• v.28 no.5
• /
• pp.569-576
• /
• 2018

This study was carried out to investigate the antioxidative, nitrite-scavenging, alcohol-metabolizing, and anti-inflammatory effects of black-cherry tomato juice (BCTJ) on LPS-induced RAW 264.7 cells. The total phenol content of the BCTJ was $156.83{\mu}g\;tannic-acid-equivalent/ml$. The antioxidative effects of BCTJ were measured using DPPH radical-scavenging activity and SOD-like assay. DPPH radical-scavenging activity of BCTJ was increased in a dose-dependent manner (p<0.05) and was 83.39% at 40%. SOD-like activity of BCTJ was 22.01% at 100%. The effects of BCTJ on alcohol-metabolism were determined by measuring generations of reduced nicotinamide adenine dinucleotides (NADH) by alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH). ADH and ALDH activities were 198.87% and 181.89% at 40%, respectively. Nitric scavenging activities of BCTJ were 85.06%, 58.25%, and 43.68% at pH values 1.2, 3.0, and 6.0, respectively, at 50%. Anti-inflammatory effects were examined in LPS-stimulated RAW 264.7 cells. Nitric-oxide production was reduced to 83.55% by the addition of BCTJ at 10%. These results suggest that black-cherry tomato juice has great potential as a resource for natural health products.

• Journal of Life Science
• /
• v.20 no.3
• /
• pp.326-335
• /
• 2010

The concentration of phenolics in Phellinus pini (CY001) extracts, expressed as mg of GAEs per g of P. pini fractions, and the EtOAc fraction (436.5 mg GAEs/g) of P. pini had a higher phenolic content than other fractions. Several biochemical assays were used to screen antioxidant properties such as reducing power, 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity, NBT/XO superoxide system and inhibition of DCF/AAPH peroxyl radicals. Among the six mushroom extracts, the EtOAc fraction from P. pini (CY001) showed the most potent DPPH radical, superoxide radical, and peroxyl radical scavenging activities, with $IC_{50}$ values of $11.49\;{\mu}g/ml$, $8.32\;{\mu}g/ml$, and $1.91\;{\mu}g/ml$, respectively. The EtOAc fraction of P. pini (CY001) significantly inhibited enzymatic lipid peroxidation and effectively attenuated LPS-induced NO production of RAW 264.7 cells without cytotoxicity. We also found that the EtOAc fraction had a significant hepato-protectant effect on tacrine-induced cytotoxicity in HepG2 cells. These findings suggest that P. pini (CY001) may have potential as a natural antioxidant, which contains compound(s) with radical scavenging activity.

• Journal of Life Science
• /
• v.27 no.9
• /
• pp.986-993
• /
• 2017

Lemon myrtle (Backhousia citriodora) has been identified as one of the plants that are likely to undergo important commercial exploitation. This study was carried out to investigate the anti-inflammatory activities and nitric oxide synthase (iNOS) expression of hot water (LMW) and 80% ethanol (LME) extracts from lemon myrtle leaf in lipopolysaccharide-induced (LPS) RAW 264.7 cells. The total phenol content of LMW and LME was 207.44 and $331.54{\mu}g$ tannic acid equivalents (TAE)/mg, respectively (p<0.01). DPPH radical scavenging activities of LMW and LME were remarkably increased in a dose-dependent manner, and were about 90.69% and 92.50% at 0.5 mg/ml, respectively. Superoxide dismutase (SOD) activities of LMW and LME were 106.22% and 103.58% at 1 mg/ml, respectively. The highest activity (91.03%) of nitrite-scavenging was observed for LME at 1 mg/ml at pH 1.2, while the activity for LMW was about 81.03% under the same conditions (p<0.05). Anti-inflammatory effect was examined in LPS stimulated RAW 264.7 cells. Nitric oxide (NO) production were reduced to 35.41% and 78.39% by addition of LMW and LME at 0.5 mg/ml, respectively (p<0.05). LMW and LME reduced protein expression of inducible nitric oxide synthase (iNOS) in a dose-dependent manner (p<0.05). These results, we conclude that lemon myrtle may be a highly valuable natural product owing to its high-quality functional components as well as its anti-oxidant and anti-inflammatory activities.

• Korean Journal of Food Science and Technology
• /
• v.46 no.5
• /
• pp.601-608
• /
• 2014

In this study, we investigated the antioxidant activity of green kohlrabi juice (GKJ) and purple kohlrabi juice (PKJ) using various in vitro methods. The results of 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging activities, ferric ion reducing antioxidant power (FRAP), reducing power, and nitrite scavenging activities showed that GKJ possessed higher antioxidant activity than PKJ. Green kohlrabi powder (GKJP) and purple kohlrabi powder (PKJP) inhibited hydrogen peroxide-induced cell death in human dermal fibroblasts. In addition, GKJP and PKJP suppressed intracellular reactive oxygen species (ROS) production induced by hydrogen peroxide in human dermal fibroblasts. These results suggest that green and purple kohlrabi juices are potential natural sources of antioxidants.