• Journal of Life Science
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• v.26 no.9
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• pp.1097-1104
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• 2016

With the development of next generation sequencing (NGS), large numbers of transcriptional molecules have been discovered. Most transcripts are non -coding RNAs (ncRNAs). Among them, long non-coding RNAs (lncRNAs) with more than 200 nucleotides represent functional RNA molecule that will not be translated into protein. In plants, lncRNAs are transcribed by RNA polymerase II (Pol II) or Pol III, Pol VI and Pol V. After transcription of these lncRNAs, more RNA processing mechanisms such as splicing and polyadenylation occurs. The expression of plant lncRNAs is very low and is tissue specific. However, these lncRNAs are strongly induced by specific external stimuli. Because different external stimuli including environmental stresses induce a large number of plant lncRNAs, these lncRNAs have been gradually considered as new regulatory factors of various biological and development processes such as epigenetic repression, chromatin modification, target mimicry, photomorphogenesis, protein relocalization, environmental stress response, pathogen infection in plants. Moreover, some lncRNAs act as precursor of short RNAs. Although a large number of lncRNAs have been predicted and identified in plants, our current understanding of the biological function of these lncRNAs is still limited and their detailed regulatory mechanisms should be elucidated continuously. Here, we reviewed the biogenesis and regulation mechanisms of lncRNAs and summarized the molecular functions unraveled in plants.

• Journal of Life Science
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• v.21 no.1
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• pp.119-126
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• 2011

There is a growing recognition of the significance of $H_2S$ as a biological signaling molecule involved in vascular and nervous system functions. In mammals, two enzymes in the transsulfuration pathway, cystathionine ${\beta}$-synthase (CBS) and cystathionine ${\gamma}$-lyase (CGL), are believed to be chiefly responsible for $H_2S$ biogenesis. Genetic inborn error of CGL leads to human genetic disease, cystathioninuria, by accumulating cystathionine in the body. This disease is secondarily associated with a wide range of diseases including diabetes insipidus and Down's syndrome. Although the human CGL (hCGL) overexpression is essential for the investigation of its function, structure, reaction specificity, substrate specificity, and protein-protein interactions, there is no clear report concerning optimum overexpression conditions. In this study, we report a detailed analysis of the overexpression conditions of the hCGL using a bacterial system. Maximum overexpression was obtained in conditions of low culture temperature after inducer addition, performing low aeration during overexpression, and using a low concentration inducer (0.1 mM, IPTG) for induction. Expressed hCGL was purified by His-tag affinity column chromatography and confirmed by Western blot using hCGL antibody and enzyme activity analysis. We also report that the His tag with TEV site attached protein exhibits 76% activity for ${\alpha}-{\gamma}$ elimination reaction with L-cystathionine and 88% for ${\alpha}-{\beta}$ elimination reaction with L-cysteine compared to those of wild type hCGL, respectively. His tag with TEV site attached protein also exhibits a 420 nm absorption maximum, which is attributed to the binding cofactor, pyridoxal 5'-phosphate (PLP).

• 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.

• Tuberculosis and Respiratory Diseases
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• v.55 no.5
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• pp.488-498
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• 2003

Background : Activation of the transcription factor NF-${\kappa}B$ has been shown to protect cells from tumor necrosis factor-alpha, chemotherapy, and radiation-induced apoptosis. NF-${\kappa}B$-dependent cIAP expression is a major antiapoptotic mechanism for that. NF-${\kappa}B$ activation and cIAP expression in A549 lung cancer cells which is relatively resistant to radiation-induced cell death were investigated for the mechanism of radioresistance. Materials and methods : We used A549 lung cancer cells and Clinac 1800C linear accelerator for radiation. Cell viability test was done by MTT assay. NF-${\kappa}B$ activation was tested by luciferase reporter gene assay, Western blot for $I{\kappa}B{\alpha}$ degradation, and electromobility shift assay. For blocking ${\kappa}B$, MG132 and transfection of $I{\kappa}B{\alpha}$-superrepressor plasmid construct were used. cIAP expression was analyzed by RT-PCR and cIAP2 promoter activity was performed using luciferase assay system. Results : MTT assay showed that cytotoxicity even 48 hr after radiation in A549 cells were less than 20%. Luciferas assay demonstrated weak NF-${\kappa}B$ activation of $1.6{\pm}0.2$ fold compared to PMA-induced $3.4{\pm}0.9$ fold. Radiation-induced $I{\kappa}B{\alpha}$ degradation was observed in Western blot and NF-${\kappa}B$ DNA binding was confirmed by EMSA. However, blocking NF-${\kappa}B$ using MG132 and $I{\kappa}B{\alpha}$-superrepressor transfection did not show any sensitizing effect for radiation-induced cell death. The result of RT-PCR for cIAP1 & 2 expression was negative induction while TNF-${\alpha}$ showed strong expression for cIAP1 & 2. The cIAP2 promoter activity also did not show any change compared to positive control with TNF-${\alpha}$. Conclusion : We conclude that activation of NF-${\kappa}B$ does not determine the intrinsic radiosensitivity of cancer cells, at least for the cell lines tested in this study.

• Journal of Life Science
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• v.21 no.5
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• pp.631-646
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• 2011

Mesenchymal stem cells (MSC) are multipotent and can be isolated from diverse human tissues including bone marrow, fat, placenta, dental pulp, synovium, tonsil, and the thymus. They function as regulators of tissue homeostasis. Because of their various advantages such as plasticity, easy isolation and manipulation, chemotaxis to cancer, and immune regulatory function, MSCs have been considered to be a potent cell source for regenerative medicine, cancer treatment and other cell based therapy such as GVHD. However, relating to its supportive feature for surrounding cell and tissue, it has been frequently reported that MSCs accelerate tumor growth by modulating cancer microenvironment through promoting angiogenesis, secreting growth factors, and suppressing anti-tumorigenic immune reaction. Thus, clinical application of MSCs has been limited. To understand the underlying mechanism which modulates MSCs to function as tumor supportive cells, we co-cultured human adipose tissue derived mesenchymal stem cells (ASC) with cancer cell lines H460 and U87MG. Then, expression data of ASCs co-cultured with cancer cells and cultured alone were obtained via microarray. Comparative expression analysis was carried out using DAVID (Database for Annotation, Visualization and Integrated Discovery) and PANTHER (Protein ANalysis THrough Evolutionary Relationships) in divers aspects including biological process, molecular function, cellular component, protein class, disease, tissue expression, and signal pathway. We found that cancer cells alter the expression profile of MSCs to cancer associated fibroblast like cells by modulating its energy metabolism, stemness, cell structure components, and paracrine effect in a variety of levels. These findings will improve the clinical efficacy and safety of MSCs based cell therapy.

• Journal of Life Science
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• v.25 no.11
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• pp.1214-1222
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• 2015

The purpose of this study was to determine the effect of Pueraria lobate-root based combination supplementation containing Rehmannia glutinosa and exercise on histone modification in ovariectomized rat hindlimb skeletal muscle. Sixty rats were fed with high fat diet and randomly assigned into the following groups for 8 weeks: 1)HSV; High fat+Sedentary+Vehicle, 2)HSP; High fat+Sedentary+PR, 3)HSH; High fat+Sedentary+Estradiol, 4)HEV; High fat+Ex+Vehicle, 5)HEP; High fat+Ex+PR, 6)HEH; High fat+Ex+Estradiol. Exercise consisted of low intensity treadmill exercise(1-4th wk:15 m/min for 30 min, 5-8th wk: 18 m/min for 40 min, 5 times/week). The result of this study showed that exercise and Pueraria and Rehmannia glutinosa intake suppressed weight gain. Furthermore, exercise and Pueraria and Rehmannia glutinosa intake increased muscle mass. This study observed H3K9 acetylation and demethylation in plantaris muscle in exercised group, but no difference in soleus muscle. To test whether the decrease in HDAC4, HDAC5 and G9a mRNA levels after exercise and Pueraria/Rehmannia glutinosa intake, HDAC4, HDAC5 and G9a mRNA levels were determined by real-time PCR. Only exercise induced HDAC5 and G9a mRNA reduction in plantaris muscle, but not in soleus muscle. In conclusion, these data demonstrates that exercise and Pueraria/Rehmannia glutinosa intake effect on body compositions. These changes are regulated by epigenetic modifications, such as histone acetylation and methylation. Future studies should focus on gene-specific epigenetics and other epigenetic mechanism for Pueraria/Rehmannia glutinosa intake.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.4
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• pp.483-490
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• 2015

The purpose of this study was to investigate the effects of 50% ethanol extracts of ripe black raspberry (Rubus occidentalis, RBR) on hypertension in spontaneously hypertensive rats (SHR). The final systolic blood pressure of the group treated with RBR for 12 weeks was significantly lower than that of the SHR group. The mRNA expression level of endothelial nitric oxide synthase (eNOS) was significantly decreased in SHR. However, treatment with RBR and captopril increased the level of eNOS mRNA in SHR. Moreover, plasma levels of homocysteine and plasminogen activator inhibitor-1 were significantly reduced by RBR. Plasma total cholesterol, high-density lipoprotein, and low-density lipoprotein cholesterol levels were lower in SHR than Wistar Kyoto rats (WKY). However, there was no significant difference in plasma triglyceride level between WKY and SHR. The number of eosinophilic cardiac muscle cells was reduced in heart muscles after treatment with captopril and RBR. Therefore, this study suggests that RBR extracts may be useful for improvement of hypertension.

• The Korean Journal of Pesticide Science
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• v.19 no.3
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• pp.279-286
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• 2015

Cotesia plutellae bracovirus (CpBV) is a polydnavirus symbiotic to C. plutellae parasitizing young larvae of the diamondback moth, Plutella xylostella. Several CpBV genes play important roles in suppressing immune responses of the parasitized larvae. This study tested a hypothesis that the CpBV genes inducing host immunosuppression could be applied to develop a potent recombinant baculovirus. Based on a previous study, a recombinant baculovirus expressing CpBV-ELP1 (AcMNPV-ELP1) was selected and multiplied using larvae of the beet armyworm, Spodoptera exigua. The recombinant viruses were produced in a yield of $5{\times}10^{10}$ polyhedral inclusion body (PIB)/larva. The cultured AcMNPV-ELP1 exhibited a much higher pathogenicity against S. exigua larvae. However, its insecticidal activity was varied among larval instars of S. exigua, in which first and late instars were high susceptible. Spray of the recombinant baculovirus ($5{\times}10^6PIB/mL$) exhibited higher control efficacy (${\approx}$ 88%) against S. exigua larvae infesting cabbage than a chemical insecticide, tebufenozide, at 7 days after treatment. These results indicate that AcMNPV-ELP1 mass-cultured using host insect system is highly pathogenic and can be applied to develop a novel microbial control agent.

• Journal of Life Science
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• v.20 no.12
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• pp.1838-1843
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• 2010

Skin aging is related to genetic and environmental factors (e.g., gene mutation and UV radiation respectively). To develop a new anti-wrinkle cosmetic or functional food by using Korean rice wine cake, we examined the effects of Korean rice wine cake, a brewery byproduct, on antioxidant effect, collagen synthesis and expression of MMP-1. Interestingly, we found that Korean rice wine cake has the ability to promote scavenging activity of DPPH radical. We also found that the cell proliferation and synthesis of collagen in HS27 cells was increased by Korean rice wine cake in a concentration-dependent manner. However, elastase inhibitory activity was not changed. In addition, the expression of MMP-1 was inhibited by Korean rice wine cake in a concentration-dependent manner. All these results suggest that Korean rice wine cake can be effectively used for the prevention of wrinkles in human skin.

• Journal of Plant Biotechnology
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• v.43 no.4
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• pp.492-499
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• 2016

This study assessed the whitening and anti-aging effects of the Cistanche deserticola extract, to develop a cosmetic substance. The cell viability of the Cistanche deserticola extract was evaluated in B16F10 melanoma cells by the MTT (3-(4,5-dimethylthaiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay. The cell viability of the extract was determined to be 90% at 4mg/ml concentration. Furthermore, the tyrosinase, collagenase, and elastase mRNA expression level were measured by RT-PCR, using the Cistanche deserticola extract treated B16F10 melanoma cells. At 4 mg/ml concentration, mRNA expression level of tyrosinase, collagenase, and elastase was dramatically decreased to 80.9%, 37.6%, and 70.9%, respectively. The antioxidant activity of the Cistanche deserticola extract was determined by DPPH free radical scavenging. The DPPH free radical scavenging capacities ranged from 70.6% to 82.6%, when evaluated from 2 mg/ml to 10mg/ml concentrations. The effects of whitening and anti-aging of the Cistanche deserticola extracts were examined at 2, 4, 6, 8, and 10 mg/ml concentration. Tyrosinase activities were inhibited from 66.8% to 78.5%, elastase activities were inhibited from 67.6% to 79.3%, collagenase activities were inhibited from 72.3% to 83.6%, and hyaluronidase activities were inhibited from 65.8% to 69.2%, respectively. These data suggest that the Cistanche deserticola extract is effective in whitening and anti-aging; therefore, it is considered to be a functional cosmetic material in cosmetic products.