• Development and Reproduction
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• v.15 no.2
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• pp.179-185
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• 2011

Some phytoestrogens in soy and red wine, for example, might have beneficiary rather than adverse effects. In particular, dietary soy intake seems to be highly correlated with protection of breast cancer, osteoporosis and cardiovascular disorders. However, questions persist on the potential adverse effects of the main soy constituent genistein (GS) on female reproductive physiology. Previously we found that prepubertal exposure to GS could activate the reproductive system of immature female rats leading to precocious puberty onset, and intracerebroventricularly (ICV) injected GS could directly activate hypothalamic kisspeptin-GnRH neuronal circuits in adult female rats. The present study was performed to examine the hypothalamus-specific GS effects in prepubertal female rats and which subtype of estrogen receptor is mediated in this GS effect. Prepubertal female rats (PND 30) were anaesthetized, treated with single dose of GS (3.4 ${\mu}g$/animal), and sacrificed at 2 hrs post-injection. To determine the transcriptional changes of reproductive hormone-related genes in hypothalamus, total RNAs were extracted and applied to the semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). ICV infusion of GS significantly lowered the transcriptional activities of mTOR (1:$0.361{\pm}0.058$ AU, p<0.001) but increased that of GAD67 (1:$1.285{\pm}0.099$ AU, p<0.05), which are known to act as an upstream modulator of kisspeptin and GnRH neuronal activities in the hypothalamus, respectively. GS administration enhanced significantly the mRNA levels of KiSS-1(1:$1.458{\pm}0.078$ AU, p<0.001), and exerted no effect on the mRNA level of kisspeptin receptor GPR-54 (1:$1.29{\pm}0.08$ AU). GnRH gene expression was significantly decreased in GS-treated group compared to control group (1:$0.379{\pm}0.196$ AU, p<0.05). There was no difference in the mRNA level of $ER{\alpha}$ in the GS-treated group compare to control group (1:$1.180{\pm}0.390$ AU, Fig. 3A). However, icv infusion of GS significantly increased the transcriptional activities of $ER{\beta}$ (1:$4.209{\pm}0.796$ AU, p<0.01, Fig. 3B). Taken together, the present study indicated that the acute exposure to GS could directly alter the hypothalamic GnRH modulating system in prepubertal female rats. Our study strongly suggested the involvement of $ER{\beta}$ pathway in GS's hypothalamus-specific action, and this idea is consistent with the GS's well-known $ER{\beta}$-mediated protective action in breast cancer.

• Horticultural Science & Technology
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• v.31 no.6
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• pp.748-755
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• 2013

This study was conducted to find a salt tolerance gene in Brassica rapa. In order to meet this objective, we analyzed data from a KBGP-24K oligo chip [BrEMD (Brassica rapa EST and microarray database)] of the B. rapa ssp. pekinensis 'Chiifu' under salt stress (250 mM NaCl). From the B. rapa KBGP-24K microarray chip analysis, 202 salt-responsive unigenes were primarily selected under salt stress. Of these, a gene with unknown function but known full-length sequence was chosen to closely investigate the gene function. The selected gene was named BrSSR (B. rapa salt stress resistance). BrSSR contains a 285 bp open reading frame encoding a putative 94-amino acid protein, and a DUF581 domain. The pSL94 vector was designed to over-express BrSSR, and was used to transform tobacco plants for salt tolerance analysis. T1 transgenic tobacco plants that over-expressed BrSSR were selected by PCR and DNA blot analyses. Quantitative real-time RT PCR revealed that the expression of BrSSR in transgenic tobacco plants increased by approximately 3.8-fold. Similar results were obtained by RNA blot analysis. Phenotypic characteristics analysis showed that transgenic tobacco plants with over-expressed BrSSR were more salt-tolerant than the wild type control under 250 mM NaCl for 5 days. Based on these results, we hypothesized that the over-expression of BrSSR may be closely related to the enhancement of salt tolerance.

• Journal of Plant Biology
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• v.32 no.1
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• pp.33-40
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• 1989

Polysomal polyadenylated mRNAs which were purified from pea leaves were fractionated by sucrose grandient sedimentation. Fractions corresponding to the peak at 11.5S were found to contain mostly mRNA encoding the precursor polypeptide to the small subunit of ribulose bisphosphate carboxylase (rbcS) by in vitro translation in wheat germ extract. Double-stranded cDNA which was synthesized from the 11.5S mRNA was cloned into Hind III site of plasmid pBR 325. A cDNA clone, H24, was identified to code for rbcS. In vitro translation product of the hybridization-selected mRNA was molecular weight 20,000, presumably the precursor of rbcS. The nucleotide sequences of the H24 showed almost complete homology with the sequences encoding the transit peptide of the rbcS-3A gene which was reported by Fluhr et al.(1986).

• Radiation Oncology Journal
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• v.19 no.4
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• pp.381-388
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• 2001

Purpose : This study was conducted to assess the effects of x-irradiation on the expression of the novel glutathione S-transferase K1 gene. Materials and methods : Human glutathione S-transferase K1 (hGSTK1) DNA was purified and ligated to a pcDNA3.1/Myc-His(+) vector for the overexpression of hGSTK1 gene. MCF-7 cells were transfected with or without the recombinant hGSTK1 gene, and irradiated with 6 MV x-ray. After incubation of 14 days, cell survival was measured and compared. The expression of hGSTK1 and the effect of x-irradiation on hGSTK1 expression were also estimated in MCF-7 cells transfected with or without the hGSTK1 gene by RT-PCR. Results : Following 2 to 12 Gy of x-irradiation, the cell survivals were higher in the MCF-7 cells transfected with the hGSTK1 gene than in those without transfection. Despite the higher cell survival in the hGSTK1-transfected cells, RT-PCR for hGSTK1 mRNA revealed no significant differences according to radiation dose, fractionation, and time after irradiation. Conclusion : The MCF-7 cells transfected with the hGSTK1 gene showed higher cell survival than those without transfection of the gene. The hGSTK1 gene might be associated with the radiosensitivity of MCF-7 cell line and further analysis should be needed.

• KSBB Journal
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• v.20 no.1 s.90
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• pp.40-45
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• 2005

The production of matrix metalloproteinases (MMPs) by the UV irradiated skin fibroblast and the degradation of exracellular matrix (ECM) by these enzymes is known as one of the main reasons of photoaging. In this study, to investigate the relationship between aging and Spatholobi caulis extract, we examined the effects of antioxidant, in vitro MMP inhibition and expression of UVA-induced MMP-1 in human dermal fibroblasts. Spathoiobi caulis extract was found to show scavenging activities of radicals and reactive oxygen species (ROS) with the $IC_{50}$ values of $45.81{\mu}g/ml$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and $3.11{\mu}g/ml$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. Spatholobi caulis extract inhibited the activities of MMP-1 in a does-dependent manner and the IC50 value calculated from semi-log plots was $31.96{\mu}g/ml$. Also, UVA induced MMP expression was reduced $74.66\%$ by treatment with Spatholobi caulis extract, and MMP-1 mRNA expression was reduced in a dose-dependent manner. Therefore Spatholobi caulis extract was able to significantly inhibit MMP expression in protein and mRNA level. All these results suggested that Spatholobi caulis extract may act as an anti-aging agent by antioxidation and reducing UVA-induced MMP-1 production.

• Journal of Marine Life Science
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• v.7 no.2
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• pp.145-153
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• 2022

Mercury (Hg) is a major concern in marine environment because of their bioaccumulation and biomagnification properties, and adverse effects to aquatic organisms at even a trace amount. However, little information on the effects of Hg, compared to other heavy metals, is available in marine small crustaceans. Here, we investigated the transcriptional modulation of metabolism-related genes in the brackish water flea, Diaphanosoma celebensis after exposure to sublethal concentration (0.2, 0.4, 0.8 ㎍/l) of HgCl₂ for 48 h. Relative mRNA expression levels of five detoxification enzyme-coding genes (cytochrome P450; cyp360A1, cyp361A1, cyp4AP3, cyp4C122, and cyp370C5) and six digestive enzyme-coding genes [alpha amylase (AMY), alpha amylase related protein (AMY-like), trypsin (TRYP), chymotrypsin-like protein (CHY), lipase (LIP), pancreatic lipase-related protein (PLRP)] were analyzed using quantitative real time reverse transcription polymerase chain reaction (qRT-PCR). As results, Hg increased the mRNA level of cyp370C5 (clan2) and cyp4AP3 (clan4) in a concentration dependent manner. A significant increase in TRYP mRNA was also concentration-dependently observed after exposure to Hg. These findings suggest that cyp370C5 and cyp4AP3 play a key role in Hg detoxification in D. celebensis, and Hg can affect energy metabolism by modulating the transcription of digestive enzyme. This study will provide better understanding the molecular effects of Hg in marine small crustacean.

• Proceedings of the Korea Information Processing Society Conference
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• 2008.05a
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• pp.433-434
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• 2008

기존의 실험을 통한 전통적인 생물학의 연구와는 달리, 미세 RNA (microRNA)의 연구에 있어 컴퓨터를 통한 프로그램 개발과 정보기술의 이용은 필수 불가결한 요소가 되었다. 컴퓨터를 바탕으로 한 대부분의 연구는 미세 RNA를 발현하는 유전자와 미세 RNA의 타겟 (target)을 예측하는 두가지 분야로 나누어 이루어지고 있다. 본 연구에서는 미세 RNA의 타겟을 예측하는 프로그램 개발시 이용되는 몇 가지 원칙들과 그 원칙들의 문제점을 서술하며, 현재 인터넷상에서 이용 가능한 프로그램들을 소개한다. 또한 컴퓨터를 통해 예측된 미세 RNA 의 타겟을 실험을 통해 검증하는 방법에 대해 논한다.

• Journal of Gastric Cancer
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• v.6 no.3
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• pp.132-138
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• 2006

Purpose: Dopamine is a neurotransmitter, but in the GIT, the roles of dopamine are a regulator of epithelial cell proliferation, an endogenous protective factor, and a regulator of stomach cancer cell proliferation. By using two different gastric-cancer cell lines, we assessed the effects of dopamine and dopamine receptors on the proliferation of human gastric-cancer cells. Materials and Methods: To assess the effects of dopamine and dopamine receptors on the proliferation of human gastric-cancer cells, we investigated cell proliferation and the expression of D1, D2L, and D2S receptor in two gastric-cancer cell lines, SNU 601 and KCU-C2. The effects of dopamine and dopamine receptors on the level of the cell proliferation were determined by staining with an A/H/E (acridine orange, hoechst and ethidium bromide) mixture. Results: After dopamine treatment, the cell viability was significantly decreased in SNU 601 cells (P<0.05) where the D2L receptor was absent, but not in KCU-C2 cells. After treatment with raclopride, a D2 receptor antagonist, dopamine-dose-dependent inhibition of cell proliferation was observed in SNU 601 cells (P<0.05). After treatment with SCH 23390, a D1 receptor antagonist, dopamine significantly increased ceil proliferation in KCU-C2 cells (P<0.05), but inhibited ceil proliferation in SNU 601 cells (no D2L receptor). Conclusion: The dopamine signal via the D1 or the D2S receptor inhibited proliferation of gastric-cancer cells, but that via the D2L receptor increased proliferation. These results suggest that the regulatory effects of dopamine in the gastric-cancer cell proliferation may be controlled by using dopamine receptors.

• Journal of Animal Science and Technology
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• v.48 no.3
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• pp.345-352
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• 2006

Korean native cattle (Hanwoo) have a good capacity to produce heavily marbled meat of high value. The intramuscular fat in Hanwoo is known to be deposit from 12 months of age by degree of slightly visible and significantly developed in 28 months of age. Lipogenesis gene expression profiling in longissimus dorsi at early and late fattening stage will be helpful to understand the mechanism of intramuscular fat deposition in skeletal muscle. Therefore, we analysed the gene expression patterns of six genes related lipid metabolism (FABP4, GLUT4, LPL, ACC, ACL and SCD) between early and late fattening stage. The mRNA expression of FABP4 at late fattening stage (27 months old) was higher about 3.0 fold than at early fattening stage (12 months old) in each three individuals of Hanwoo. However, GLUT4 mRNA expression was not different at late fattening stage compared with at early fattening stage. On the other hand, The expression patterns of LPL, ACC, ACL and SCD genes related lipid metabolism were significantly over-expressed about 3.5 fold, 2.7 fold, 3.7 fold and 7.5 fold at late fattening stage, respectively. Thus, these results suggested that lipogenesis in skeletal muscle at late fattening stage is due to increasing uptake of fatty acid by FABP4 and lipogenesis gene expression such as LPL, ACC, ACL and SCD.

• Environmental Mutagens and Carcinogens
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• v.24 no.2
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• pp.73-78
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• 2004

In order to understand the mechanism of the regulation of drug metabolizing enzyme gene expression, we have studied the induction of CYP1A1 and GST$\alpha$, $\mu$, $\pi$ enzymes in Japanese monkey and rhesus monkey after the treatment with 3-methylcholanthrene (3MC) and di-n- butyl phthalate (DBP) and bisphenol A (BPA). The levels of mRNA were measured by RT-PCR in brain, intestine and liver. In the case of adult monkey, treatment with 3MC induced CYP1A1 mRNA in liver by 10-fold. The treatment with DBP induced CYP1A1 mRNA. Effects of 3MC and DBP on GST mRNA expression was not clear. But GST$\mu$ was slightly inhibited by the treatment with 3MC and DBP. GST$\pi$ was not induced by the treatment with 3MC and DBP in liver. GST$\alpha$ was slightly induced by the treatment with 3MC and DBP in liver. In the case of fetus monkey, the basal levels of fetus CYP1A1 mRNA and GSTs mRNA were relatively low compared to adult monkey. As the age of monkey increased, the basal levels of CYP1A1 mRNA were also increased. 3MC induced the expression of CYP1A1 mRNA in liver. The levels of GST$\mu$ and GST$\alpha$ were not changed by the treatment with 3MC and DBP. GST$\pi$ was slightly induced by the treatment with 3MC and DBP.