• The Korean Journal of Pesticide Science
• /
• v.8 no.2
• /
• pp.95-102
• /
• 2004

The present study was conducted to test and evaluate estrogenic effect of 17 pesticides including benomy1 and carbaryl, being suspected as endocrine disrupting chemicals. For estrogenic effect examination, luciferase assay were achieved with human ovarian cancer cell, BG1Luc4E2. Estrogenic effects of cypermethrin, dicofol, endosulfan, esfenvalerate, and fenvalerate were observed at the concentration of $10^{-5}$ M by estrogen receptor binding assay. Relative luciferase potency and relative luciferase effects compared with $10^{-10}$ M 17 $\beta$-estradiol were $10^{-5}$, 56% for dicofol, and $10^{-5}$, 72% for endosulfan, respectively. Estimated maximum daily intake for pesticides was calculated from maximum residue limit of agricultural commodity and food consumption was 1.2298 mg/person. Theoretically calculated blood estrogen level from dietary intake for pesticides based on MRL in Korea, 3.075 ng/L was equivalent to 15% of estrogen concentration in normal blood, but practical monitoring data, 0.01938 ng/L was equal to 0.09693% of estrogen concentration in normal blood.

• Journal of the korean academy of Pediatric Dentistry
• /
• v.29 no.3
• /
• pp.337-344
• /
• 2002

Runx2, a Runt-related osteoblast-specific transcription factor, is essential for osteoblast differentiation and function. Runx2 was identified as a key regulator of osteoblast-specific gene expression through its binding to the OSE2 element present in these genes. However, little is known about the signaling mechanism regulating Runx2 activity. This study examines the role of protein kinase C (PKC) pathway and mitogen-activated protein kinase (MAPK) pathway in regulating Runx2 and bone marker genes (osteopontin; OP, osteocalcin; OC). Luciferase assay and Northern blot analysis suggested that the stimulation of PKC by PMA increased transcription activity of Runx2 and bone marker genes (OP and OC) and also increased expression of Runx2. The stimulation of MAPK by okadaic acid increased transcription activity of Runx2 and bone marker genes (OP and OC). Pretreatment with PD98059 (Erk pathway inhibitor) and SB203580 (P38 pathway inhibitor) prior to PMA treatment decreased PMA stimulated Runx2 activity. Together these results indicate that both PKC and MAPKs are involved in the regulation of Runx2 activity and also the stimulation of Runx2 transcriptional activity by the PKC pathway is through activation of MAPK pathway.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.39 no.1
• /
• pp.22-28
• /
• 2019

We previously isolated 9 clones that show stronger signal compared to Bombyx mori cytoplasmic actin gene (BmA3) by using a dot blot hybridization. In this study, we focused on one clone among these clones which has high amino acid similarity with ${\beta}$-tubulin gene of B. mori. This clone was ubiquitously expressed in all tissues and developmental stage of B. mori. As result of promoter assay using dual luciferase assay system, we found the highest transcription activity region (-750/-1) in the 5'-flanking region of ${\beta}$-tubulin gene, which has about 47 fold more intensive promoter activity than BmA3 promoter. Moreover, the ${\beta}$-tubulin promoter was normally regulated in Bm5, Sf9, and S2 cells. Therefore, we suggest that ${\beta}$-tubulin promoter may be used more powerful and effectively for transgene expression in various insects containing B. mori as a universal promoter.

• The Journal of Korean Medicine
• /
• v.29 no.3
• /
• pp.88-99
• /
• 2008

Objectives: In order to identify the anti-inflammatory and analgesic properties of Salvia miltiorrhiza (Dan-Sam), widely used in Korean traditional medicine, an in vitro screening system was designed using pGL3, a luciferase reporter vector, and the tumor necrosis factor (TNF)-${\alpha}$ and cyclooxygenase (COX)-2 as target genes. Methods: The promoter regions of each gene were generated by PCR using the human chromosome as template DNA, and inserted into pGL3 vector with Kpn I and Hind III. The final construct was transfected into human myelomonocytic leukemia cells (U-937) that could be differentiated and activated by phorbol 12-myristate 13-acetate (PMA) or lipopolysaccharide (LPS). Using this system, the anti-inflammatory and analgesic effects of several herbal extracts regarded to have the medicinal effects of diminishing body heat and complementing Qi were tested. The chemicals PD98059 and berberine chloride were used as controls of the transcriptional inhibitors of TNF-${\alpha}$ and COX-2, respectively. Results: Salvia miltiorrhiza (Dan-Sam) demonstrated significant decrease of TNF-${\alpha}$ and COX-2 mRNA in the in vitro assay system. In MTT assay, Salvia miltiorrhiza (Dan-Sam) did not significantly inhibit the survival and proliferation of human myelomonocytic leukemia cells (U-937). Conclusions: Salvia miltiorrhiza (Dan-Sam) was found to exhibit the significant medicinal properties of anti-inflammatory and analgesic effects.

• Journal of Aquaculture
• /
• v.11 no.4
• /
• pp.457-463
• /
• 1998

An expression vector, pUC19N6-luc, containing nuclear matrix attachment region(MAR) isolated from Misgurnus mizolepis liver and control expressino vector, pUC19-luc, were constructed. After these vectors were transferred into CHSE-214 cell line by electroporation, the expression rate of luckferase gens, copy number of vectors and chromosome integration of vectors were analyzed by using assay of luciferase activity, PCR and Southern blotting. While the expression pattern of luciferase gene of pUC19-luc was shown in typicla transient ecpression pattern, that of pUC19N6-luc was highly increased at the 5 days after transfectrion. Although the cope number of pUC19N6-luc vector was higher than that of pUC19-luc vector, these vectors were integrated into chromosome at the same time point in the transfected CHSE-214 cells. In conclusion, the increase of luciferase gene expression of pUC19N6-luc was resulted from not the maintaining of the high copy number but the formation of transcription-favorable structure by MAR effect after chromosomal integration.

• Proceedings of the Korean Society of Toxicology Conference
• /
• 2002.11b
• /
• pp.164-164
• /
• 2002

The estrogenic activity of 47 plant extracts was assessed by reporter gene assay using MCF-7 breast cancer cell lines stably transfected with luciferase reporter gene. The estrogenic activity of food extracts was expressed as 17${\beta}$-estradiol(E2) equivalent concentration(EEQ), the concentration of E2 that resulted in the same relative luciferase unit(RLU) of the food extract of 0.2mg/$m\ell$.(omitted)

• Journal of the Korean Chemical Society
• /
• v.55 no.3
• /
• pp.400-404
• /
• 2011

Safflower is called as the 'beneficial flower' because 'it helps human health', and it was introduced as red flower in Tonguibogam due to the red color of floral leaf. From old times, it has been used for the material of cloth and rouge. Recently, polyphenol compound, the main ingredient of safflower, known as anti-aging and anti-oxidizing material in the healthy food industry becomes the emerging hot topic. This study aims to confirm by DDT (Disk Diffusion Test) assay, MTT assay, and NF-${\kappa}$B Luciferase activity inhibition assay in vitro that polyphenol compound, which is the main ingredient of safflower, has the anti-microbial efficacy to inhibit the growth of acne germs that make troubles for the teenagers or middle aged. Also it aims to evaluate its clinical efficacy on the acne skin, utilizing the facial cleansing cosmetic form of soap sample. This study can contribute to take a major step forward to the development of cosmetic soap for acne in the cosmeceutical industry.

• Proceedings of the Korean Society of Applied Pharmacology
• /
• 2003.11a
• /
• pp.88-88
• /
• 2003

Cytochrome P-450 3A4 (CYP3A4) is major enzyme in human liver, the role of this is detoxification and metabolizing more than 50% clinical drugs in use. Expression of CYP3A4 is transciptionally regulated by the Pregnenolone X receptor (PXR), of which human form is Steroid and Xenobiotics receptor (SXR). SXR is activated by wide range of endogenous and exogenous compounds, and then induces CYP3A4 gene expression. In the previous study, it has been known that proximal promoter (-864 to +64) does not response to chemical inducers such as pregnenolone 16a-carbonitrile (PCN), Rifampicin, Estrogen in terms of transcription of CYP 3A4 in cultured cells. Here, we developed luciferase reporter gene assay system to detect SXR-based CYP 3A4 transcriptional activity. We have used CYP3A4-Luc plasmid that contains proximal promoter of human CYP3A4 gene upstream of the luciferase gene. We did transient transfection of 3A4-luciferase gene and SXR. In the HepG2 cells transfected with CYP3A4-Luc, when rifampicin treatment was combined with histone deacetylase inhibitor (HDAC Inhibitor), such as Trichostatin A, Hc-toxin and IN 2001 of the luciferase activity was induced 10-20 fold over control.

• Proceedings of the Korea Society of Environmental Toocicology Conference
• /
• 2003.10a
• /
• pp.178-178
• /
• 2003

Cytochrome P-450 3A4 (CYP3A4) is major enzyme in human liver, the role of this Is detoxification and metabolizing more than 50% clinical drugs in use. Expression of CYP3A4 is transciptionally regulated by the Pregnenolone X receptor (PXR), of which human form is Steroid and Xenobiotics receptor (SXR). SXR is activated by wide range of endogenous and exogenous compounds, and then induces CYP3A4 gene expression. In the previous study, it has been known that proximal promoter (-864 to ＋64) does not response to chemical inducers such as pregnenolone 16a-carbonitrile (PCN), Rifampicin, Estrogen in terms of transcription of CYP 3A4 in cultured cells. Here, we developed luciferase reporter gene assay system to detect SXR-based CYP 3A4 transcriptional activity. We have used CYP3A4-Luc plasmid that contains proximal promoter of human CYP3A4 gene upstream of the luciferase gene. We did transient transfection of 3A4-luciferase gene and SXR. In the HepG2 cells transfected with CYP3A4-Luc, when rifampicin treatment was combined with histone deacetylase inhibitor (HDAC Inhibitor), such as Trichostatin A, Hc-toxin and IN 2001 of the luciferase activity was induced 10-20 fold over control.

• The Korean Journal of Zoology
• /
• v.39 no.4
• /
• pp.378-386
• /
• 1996

We have developed a couple of new luciferase reporter plasmida with very low background reporter actlvltlea. One can be used to measure the promoter strength, after insertion of some promoter fragment into the reporter plasmid, and the other, with very low basal promoter actlvltlea, allis In studying eukaryotic transcriptional regulators. The latter reporter plasmid contains such cli elements as a 17 nucleotide long inftlator, Spl.blndIng sftes, GAL4 binding sltea, and bInding sitea for a certain Drosophila homeodomain proteins. In an attempt to construct an improved reporter plaimid by fadlltating transcriptional termination and minimizing any interference by cryptic promoters which may be preaent in the reporter pleamld DNA, we have inserted transsrlptional termination-related signals, a three tandem repeat of SV4O polyadenylatlon signal (AAA) and the putative transcrtptional termination signal (UMS) of the mouse c-mos gene, Into just upstream of the initIator, and the promoter actlvitiea were measured by a transIent expression assay employing the Drosophila Schneider line 2 cells. As expected, the basal promoter activitIes decreased maximally when both transcription termination related elements were inserted. Moreover, the reporter plasmld with the two elements allowed more sensitive measurement of transcriptional activation than the reporter piasmid without them. Theae reporter plasmids can be used for studying transcriptional regulators of higher organisms Including mammals as well as Droiophlla melanogaiter.