• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.33 no.6
• /
• pp.583-590
• /
• 2007

Anticancer effect of methanol extract of Caesalpinia sappan L. on oral carcinoma(KB) and osteosarcoma(HOS) cells were investigated in this study. In order to elucidate the anticancer mechanism of Caesalpinia sappan L, we analyzed telomerase inhibitory effect of the methanol extract of Caesalpinia sappan L. In addition, we prepared 5 fraction samples according to its polarity differences and analyzed anticancer effects on oral carcinoma and osteosarcoma cells. Following results are obtained in this study. 1. 50% cell proliferation inhibitory value($IC_{50}$) of the methanol extract of Caesalpinia sappan L. against oral carcinoma(KB) cells and osteosarco ma(HOS) cells were $9.0{\mu}g/ml\;and\;10.9{\mu}g/ml$, respectively. 2. The methanol extract of Caesalpinia sappan L. showed inhibitory effect of telomerase which is required for cancer cell immortality. Therefore, it seems that the anticancer effect of methanol extract of Caesalpinia sappan is at least partially due to telomerase inhibitory effect. 3. Five fraction samples were prepared according to its polarity and 88.7% of ingredient of total methanol extract was transferred to ethylacetate fraction. Thin layer chromatography analysis showed that dichloromethane fraction contained ingredient with relatively high polarity and ethylacetate fraction contained similar ingredient found in total methanol extract. 4. Anticancer effect was observed in n-hexane, dichloromethane, and ethylacetate fractions. The highest anticancer effect was found in dichloromethane fraction which had $IC_{50}$ value of $4.4{\mu}g/ml$ and > $4.0{\mu}g/ml$ against oral carcinoma(KB) cells and osteosarcoma(HOS) cells, respectively.

• Toxicological Research
• /
• v.13 no.4
• /
• pp.311-316
• /
• 1997

This study was carried out to develop antitumor effect of the n-butanol soluble fraction of Perilla frutescens on (KB cells) human oral epitheloid carcinoma cells. The cytotoxictty of methanollc extract of Perilla frutescens on KB cells was evaluated by 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide(MTT) assay. The antitumor activity of various fractions obtained from n-butanol soluble fraction of Perilla frutescens was evaluated in human oral epithelold carcinoma cells. The antitumor acavity of the n-butanol soluble fraction on human oral epitheloid carcinoma cells was evaluated by MTT assay of colorimetric method. The light microscopic study was carried out to observe morphological changes of cultured human oral epitheloid carcinoma cells. These results were obtained as follows; 1. The fractions 1,2 and 3 of the n-butanol soluble fraction of Perilla frutescens were shown significant antitumor activities. 2. The number of human oral epitheloid carcinoma cells were decreased and tend to form cell cluster by treatment with fractions 1,2,3 and 4 of the n-butanol soluble fraction of Perilla frutescens. 3. The fraction 1 of the n-butanol soluble fraction of Perllla frutescens showed the highest antitumor activity on Perilla frutescens. It has been selected as a lead fraction for further examinations.

• Journal of Korean Academy of Oral and Maxillofacial Radiology
• /
• v.29 no.1
• /
• pp.105-117
• /
• 1999

Purpose: Ionizing radiations have been reported as an apoptosis initiating stimulus in various cells and it has established that sustained elevations in [Ca/sup 2+/] can lead to DNA fragmentation by Ca/sup 2+/-dependent endonucleases, ultimately resulting in apoptotic cell death. The previous experiments have been reported by using primarily thymocytes and lymphocytes and the change of [Ca/sup 2+/] was measured only by minutes or hours respectively. We need to evaluate [Ca/sup 2+/] in both several minutes and hours after irradiation of radiation of radiation therapy and verify the apoptotic cells. Materials and Methods: We have measured [Ca/sup 2+/] in human gingival epitheloid cancer cell with 10Gy irradiation, at minutely intervals and hourly intervals using digitized video-intensified fluorescence microscopy and the fluorescent Ca/sup 2+/ indicator dye, fura-2. In order to find out that the transient rise in [Ca/sup 2+/] could induced apoptosis, cells were incubated for 1 hour at 37℃ with TdT enzyme, rinsed and resuspended containing fluorescence and observed under a confocal fluorescence microscope. MTT assay was done to determine cell activity and LDH assay was done to determine the amount of necrotic cells. Results: After irradiation, the transient and temporal increasing of [Ca/sup 2+/] in the KB cells was founded. Though, there was no change in the intracellular [Ca/sup 2+/] at 30 minutes and 2 hours after irradiation. We could detect of DNA fragmented cells at 4 hours after 10Gy irradiated cells. There were no significant differences between 4 hour, 1 day, 3 day cells. There were no significant differences in MTT and LDH assay between the irradiated group and the control group after 4 hours and 1 day. Though after 3 days there were differences in MTT and LDH assay between the irradiated group was significantly decreased than the control group, in LDH assay the number of necrotic cell death of the irradiated was higher than the control group. Conclusion: In KB cells there were incipient and temporal increasing of the [Ca/sup 2+/] with 10Gy irradiation and the apoptosis was founded from 4 hours later which was earlier than seeing of the change of the amount of the cellular ability and necrosis.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.35 no.5
• /
• pp.287-293
• /
• 2009

Cell survival is the result of a balance between programmed cell death and cellular proliferation. Cell membrane receptors and their associated signal transducing proteins control these processes. Of the numerous receptors and signaling proteins, epidermal growth factor receptor (EGFR) is one of the most important receptors involved in signaling pathways implicated in the proliferation and survival of cancer cells. EGFR is often highly expressed in human tumors including oral squamous cell carcinomas, and there is increasing evidence that high expression of EGFR is correlated with poor clinical outcome of common human cancers. Therefore, we examined the antiproliferative activity of gefitinib, epidermal growth factor receptor tyrosine kinase inhibitor (EGFR TKI), in head and neck cancer cell lines. SCC-9, KB cells were cultured and growth inhibition activity of gefitinib was measured with MTT assay. To study influence of gefitinib in cell cycle, we performed cell cycle analysis with flow cytometry. Western blot was done to elucidate the expression of EGFR in cell lines and phosphorylation of EGFR and downstream kinase protein, Erk and Akt. Significant growth inhibition was observed in SCC-9 cells in contrast with KB cells. Also, flow cytometric analysis showed G1 phase arrest only in SCC-9 cells. In Western blot analysis for investigation of EGFR expression and downstream molecule phosphorylation, gefitinib suppressed phosphorylation of EGFR and downstream protein kinase Erk, Akt in SCC-9. However, in EGFR positive KB cells, weak expression of active form of Erk and Akt and no inhibitory activity of phosphorylation in Erk and Akt was observed. The antiproliferative activity of gefitinib was not correlated with EGFR expression and some possibility of phosphorylation of Erk and Akt as a predictive factor of gefitinib response was emerged. Further investigations on more reliable predictive factor indicating gefitinib response are awaited to be useful gefitinib treatment in head and neck cancer patients.

• Microbiology and Biotechnology Letters
• /
• v.22 no.6
• /
• pp.593-598
• /
• 1994

A bacterium KL-57 which exhibited biosurfactant activity was isolated. This bacterium was identified as Bacillus subtilis. The biosurfactant-producing gene of B. subtilis KL-57 was cloned into R subtilis MI113 by using plasmid pTB523. The plasmid DNA from the clone was found to carry a 18 kb PstI insert. The biosurfactant-producing gene was cleaved into 4 fragments by SmaI, 3 fragments by PvulI or EcoRl, 4 fragments by PvulI and EcoRI double digestion, 5 fragments by AccI, and 2 fragments by KpnI, HindIII or BamHI. By subcloning the 18 kb Pstl insert, a 2.3 kb EcoRl fragment conferred the biosurfactant producing activity on B. subtilis cells. The 2.3 kb had one HindIII cleave site. But Two fragments, which corresponds HindIII/EcoRl termini, exhibited no biosurfactant activity.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.17 no.6
• /
• pp.1427-1432
• /
• 2003

This study was carried out to investigate the antimicrobial and anticancer effects of Galla Rhois (GR) on pathogens isolated from oral and KB human oral epidermoid carcinoma cells. Their antimicrobial activities were tested against Streptococcus mutans (S. mutans), Staphylococcus aureus (S. aureus), Enterobacter aerogenes (E. aerogenes), Escherichia coli (E. coil) and Candida albicans (C. albicans). GR powder has the antimicrobial activity against C. albicans, S. mutans and S. aureus. The extracts of water and ethanol have the antimicrobial activity against S. sureus and C. albicans. The water extract showed inhibitory activity against the growth and pH of above mentioned reference microorganisms. The water extract of GR declined cell viability in a dose dependent manner. DNA flow cytometric analysis showed that population of sub-G/sub 0//G₁, phase of cell cycle was increased by GR extract treatment in a dose dependent manner. Western blot analysis revealed that Caspase-3 was reduced by GR extract treatment. These result suggested that GR has the effect of antimicrobial on pathogens isolated from oral, and also, has anticancer effect that associated with the induction of apoptosis in a dose dependent manner in KB human oral epidermoid carcinoma cells. It may be GR-induced apoptosis was mediated via activation of Caspase-3.

• Macromolecular Research
• /
• v.17 no.6
• /
• pp.403-410
• /
• 2009

Cytotoxicity is a severe problem of cadmium sulfide nanoparticles(CSNPs) for use in biological systems. In the present study, mercaptoacetic acid-coated CSNPs were conjugated with bovine serum albumin (BSA) to improve biocompatibility. The surface properties of the CSNPs and albumin-conjugated CSNPs (ACSNPs) were characterized by XRD, UV, FTIR, EA, TEM and DLS. Human breast cancer cells (KB cells) were then cultured in the presence of the nanoparticles to evaluate the cytotoxicity of CSNPs and ACSNPs. Finally, the fluorescence intensity of the nanoparticles' aqueous solution was examined using a fluorescence spectrometer. The results showed that the cell compatibility and fluorescence intensity of ACSNPs were higher than those of CSNPs. The strongly luminescent features of the biocompatible ACSNPs are promising for use in biological fields such as cellular labeling, intracellular tracking and molecular imaging.

• BMB Reports
• /
• v.28 no.3
• /
• pp.216-220
• /
• 1995

cDNA for human cholesteryl ester transfer protein (CETP), a potent atherogenic plasma protein that redistributes the neutral lipids among lipoproteins, was expressed in recombinant vaccinia virus-infected cells (CV-1). Two insertion vectors regulated by different promoters were constructed. The vectors were introduced into human thymidine kinase-negative ($TK^-$) 1438 cells infected with wild-type vaccinia virus (WR strain). Recombinant viruses were selected with 5-bromodeoxyuridine (BUdR) and X-gal and identified with DNA dot blot analysis (vSC11-CETP and vTM1-CETP). The CETP cDNA insert in the recombinant vaccinia virus genome was identified by Southern blot analysis. Transcription of CETP cDNA in CV-1 cells infected with recombinant vaccinia virus was monitored by Northern blot analysis using the CETP cDNA as a probe. Positive signals were detected at 1.8 kb in cells infected with vSC11-CETP and at 2.3 kb in cells infected with vTM1-CETP. The recombinant vaccinia virus-infected CV-1 cells were shown to produce functional CETP when the culture medium was subjected to the CETP assay.

• Polymer(Korea)
• /
• v.35 no.3
• /
• pp.210-215
• /
• 2011

Rutin (R) exhibits a wide range of biological activities including anticarcinogenic, antiinflammatory and antiviral actions. The purpose of this study is to investigate the effect of rutin concentrations (1 and 3 wt%) on the antibacterial activity of poly(3-hydroxybutylate-co-hydroxyvalerate) (PHBV) scaffolds. Antibacterial activity was evaluated by using Staphylococcus aureus and Klebsiella pneumoniae. Furthermore, the qualitative ongrowth of human KB endothelial cells was done to study in vitro cytotoxicity of the scaffolds. As the results, PHBV scaffolds containing 3 wt% rutin completely inhibited the proliferation of Staphylococcus aureus and Klebsiella pneumoniae. In addition, the PHBV/R scaffolds used in this study did not show any cytotoxicity when evaluated them with KB endothelial cells.

• Journal of Life Science
• /
• v.13 no.4
• /
• pp.522-528
• /
• 2003

The RAD4 gene of Saccharomyces cerevisiae is essential for the incision step of UV-induced excision repair. A yeast RAD4 gene has been previously isolated by functional complementation. In order to identify the RAD4 homologous gene from fungus Coprinus cinereus, we have constructed cosmid libraries from electrophoretically separated chromosomes of the C. cinereus. The 13 C. cinereus chromosomes were resolved by pulse-field gel electrophoresis, hybridized with S. cerevisiae RAD4 DNA, and then isolated homologous C. cinereus chromosome. The insert DNA of the RAD4 homolog was contained 3.2 kb. Here, we report the characterization of fungus C. cinereus homolog of yeast RAD4 gene. Southern blot analysis confirmed that C. cinereus contains the RAD4 homolog gene and this gene exists as a single copy in C. cinereus genome. When total RNA isolated from C. cinereus cells was hybridized with the 1.2 kb PvuII DNA fragment of the S. cerevisiae RAD4 gene, a 2.5 kb of transcript was detected. In order to investigation whether the increase of transcripts by DNA damaging agent, transcripts levels were examined after treating the cells. The level of transcript did not increase by untraviolet light (UV). This result indicated that the RAD4 homologous gene is not UV inducible gene. Gene deletion experiments indicate that the RAD4 homologous gene is essential for cell viability.