• Journal of Periodontal and Implant Science
• /
• v.30 no.2
• /
• pp.389-405
• /
• 2000

The purpose of this study is to evaluate the effect of IGF-I for DNA synthetic activity and the mRNA expression of bone matrix protein, type I collagen and osteopontin in prolifetation and differentiation of MC3T3-E1 cells. To evaluate DNA synthetic activity, cells were seeded at $2{\times}10^4cells/ml$ in 24 well plates and to evaluate mRNA of type I collagen and osteopontin cells were seeded at $5{\times}10^5cells/ml$ in 100mm culture dishes. These cells were cultured in alpha-minimum essential medium(${\alpha}-MEM$) containing 10% fetal bovine serum at $37^{\circ}C$, 5% $CO_2$ incubator. For DNA synthetic activity test 1, 10, 100ng/ml IGF-I were added to the cells which had been cultured for 3 days before 24 hours. For type I collagen mRNA expression 1, 10ng/ml IGF-I were added to the cells which had been cultured for 5, 10 days and for osteopontin mRNA expression 0.1, 1, 10ng/ml IGF-I were added to the cells which had been cultured for 5, 15, 20 days. Cell proliferaton was measured by the incorporation of [$^3H$]-thymidine into DNA and expression for type I collagen and osteopontin were measured by northern blot analysis. The results were as follows : DNA synthetic activity were generally higher in experimental group than control group. Expressions of type I collagen mRNA were higher at 5 day group and much lower at 10 day group in the control groups. In the experimental groups, mRNA expressions were slightly increased when 1 ng/ml IGF-I were added to 5 day group and decreased in all experimental 10 day groups. Expressions of osteopontin mRNA were higher at 20 day groups and lower at 15 day groups than the control groups. In the experimental groups, mRNA expressions were incereased when 0.1, 1 ng/ml IGF-I were added to 5 day group and in all the 15 day groups, but decreased when 0.1, 1, 10 ng/ml IGF-I were added to 20 day groups. IGF-I stimulated DNA synthetic activity of MC3T3-E1 cells during proliferation stage significantly, did not greatly changed effects on type I collagen mRNA expression and stimulated osteopontin mRNA expression at 15 day especially. In conclusion, we suggests that IGF-I have a tendency of stimulation effect of DNA synthetic activity but do not stimulate type I collagen mRNA in proliferation stage of MC3T3-E1 cell cultures, and stimulate osteopontin mRNA in differentiation stage of MC3T3-E1 cell cultures.

• Korean Journal of Microbiology
• /
• v.53 no.1
• /
• pp.49-54
• /
• 2017

Sus1 / ENY2 is a tiny conserved protein that is involved in chromatin remodeling and mRNA biogenesis. Sus1 is associated to two nuclear complexes, the transcriptional coactivator SAGA and the nuclear pore associated TREX2. In fission yeast, Schizosaccharomyces pombe, ortholog of Sus1 / ENY2 was identified from the genome database. Tetrad analysis showed that the S. pombe sus1 is not essential for growth. However, deletion of the sus1 gene caused cold-sensitive growth retardation with slight accumulation of $poly(A)^+$ RNA in the nucleus. And the Sus1-GFP protein is localized mainly in the nucleus. Yeast two-hybrid analysis and co-immunoprecipitation experiment showed that Sus1 interacts with Sac3, another subunit of TREX2 complex. These results suggest that S. pombe Sus1 is also involved in mRNA export from the nucleus as a component of TREX-2 complex.

• Genomics & Informatics
• /
• v.10 no.1
• /
• pp.40-43
• /
• 2012

Recent genomewide association studies of large samples have identified genes that are associated with blood pressure. The Global Blood Pressure Genetics (Global BPgen) and Cohorts for Heart and Aging Research in Genome Epidemiology (CHARGE) consortiums identified 14 loci that govern blood pressure on a genomewide significance level, one of which is $CASZ1$ confirmed in both Europeans and Asians. $CASZ1$ is a zinc finger transcription factor that controls apoptosis and cell fate and suppresses neuroblastoma tumor growth by reprogramming gene expression, like a tumor suppressor. To validate the function of $CASZ1$ in blood pressure, we decreased $Casz1$ mRNA levels in mice by siRNA. $Casz1$ siRNA reduced mRNA levels by 59% in a mouse cell line. A polyethylenimine-mixed siRNA complex was injected into mouse tail veins, reducing $Casz1$ mRNA expression to 45% in the kidney. However, blood pressure in the treated mice was unaffected, despite a 55% reduction in $Casz1$ mRNA levels in the kidney on multiple siRNA injections daily. Even though $Casz1$ siRNA-treated mice did not experience any significant change in blood pressure, our study demonstrates the value of $in$ $vivo$ siRNA injection in analyzing the function of candidate genes identified by genomewide association studies.

• Journal of Environmental Health Sciences
• /
• v.32 no.6
• /
• pp.566-570
• /
• 2006

Exposure to ultraviolet B(UVB) radiation causes skin inflammation such as pigmentation and the induction of cyclooxygenase-2(COX-2) gene expression. In this study, we investigated the effect of natural extracts from Tea, EGb 761 and Korean red ginseng(KRG), on the pigmentation and expression of COX-1 and COX-2 mRNA in UVB-irradiated C57BL/6 mice. Before UVB irradiation, the skin color was significantly showed the lightening effect by topical application of natural compounds (p<.05). In the case of UVB irradiated mice, we observed a decrease in pigmentation by compounds (p<.05). In irradiated skin, COX-1 mRNA expression is not changed following UVB irradiation, but COX-2 gene increases. Also, natural compounds lowered mRNA levels of COX-2. Therefore, these results suggest that COX-2 mRNA increases by UVB irradiation. Also, Tea, EGb 761 and KRG as a topical application may inhibit skin pigmentation and modulate COX-2 mRNA level.

• Journal of Embryo Transfer
• /
• v.17 no.3
• /
• pp.171-177
• /
• 2002

Glucocorticoid is activating mammary gland cells for lactating animals, resulting in increasing abilities of the milk synthesis. Expression of the prolactin receptor(PRL-R) in mammary gland cells was closely associated with milk production. To increase lactation ability for the Korean Native Goats at mid-lactation period. 0.05, 0.1. and 0.2 g of hydrocortisone was administrated with 5 $m\ell$ of saline. and injected into vein. For the control, 5 $m\ell$ of saline was administrated in to vein. After 24 H, the mammary gland tissue was collected, and mRNA expression rates were investigated for the alpha-casein and PRL-R using competitive PCR(polymerase chain reaction). There was no significant differences between treatment and control groups for the mRNA expression rate of PRL-R in mammary gland cells after 24 h of administration of hydrocortisone. The rate of mRNA expression for the alpha-casein was increased 37%, 630%, and 380% at 0.05, 0.1, and 0.2 g of hydrocortisone administration groups, respectively, comparing with control group. The results suggested that PR L-R mRNA expression of mammary gland cell by administration of hydrocortison was not significant, but increase of the alpha-casein mRNA expression my be differences of expression of functional proteins in the cell and expression patterns of protein secretion time to out of the cell. This study showed increase of alpha-casein mRNA expression by administration of hydrocortisone at mid-lactation period of Korean native goat.

• Tuberculosis and Respiratory Diseases
• /
• v.50 no.5
• /
• pp.579-590
• /
• 2001

Background : The aging process may be induced, at least in part, by reactive oxygen species(ROS). It has been thought that the lung could be a good source of ROS because it has a high oxygen tension. In the present study, we invetigated the inducibility of the first and last lines against oxidative stress, superoxide dismutases(CujZn-SOD and Mn-SOD) as a scavenger of ${O_2}^-\;{\cdot}$ and metallothionein(MT) as a scavenger of $OH{\cdot}$, respectively, in mouse lungs with age. Methods : Oxidative stress was induced by paraquat, an intracellular superoxide generator, at 1, 4, 8, and 12 months of age and then SODs and MT mRNAs were determined by RT-PCR method. Results : The steady-state level of Mn-SOD mRNA increased from 1 to 8 months but decreased thereafter. However, Mn-SOD mRNA was not induced by paraquat after 1 month. On the other hand, there was no change in the steady-state level of Cu/Zn-SOD mRNA, which decreased abruptly at 12 months of age. Additionally, Cu/Zn-SOD mRNA was not induced by paraquat at any age. There was no change in the steady-state level of MT mRNA with age whereas its inducibility by paraquat was intact at all ages. Conclusion : These results indicate that lack of induction of SODs with age may be one of the causative factors in the aging process while induction of MT may play an important role in the defense against oxidative stress. It is therefore implicated that the tissue antioxidant/prooxidant balance could be one of determinants of mean life span.

• 한국체육학회지인문사회과학편
• /
• v.55 no.4
• /
• pp.507-516
• /
• 2016

The purpose of this study is to identify the effects of PPARβ/δ over-expression on PGC-1α mRNA and protein stability after single bout of swimming exercise in mice skeletal muscle. Empty vector (EV) or PPARβ/δ was over-expressed in tibialis anterior(TA) using electroporation(EPO) technique to compare with non-treatment muscle(control; Con). TA muscles were dissected at 0h, 24h or 54h after termination of exercise. PGC-1α mRNA in Con, EV and PPARβ/δ over-expressed muscles were increased 6.8 fold (p<.001), 6.2 fold(p<.001) and 7.1 fold(p<.001), respectively, than sedentary(Sed) group at 0h after exercise and then reverted to Sed group levels at 24h and 54h after termination of exercise. PGC-1α and PGC-1α ubiquitination in EV treated muscles were increased 2.2 fold and 1.74 fold, respectively, than Sed group at 24h after termination of exercise, and then reverted to Sed group levels at 54h after termination of exercise. PGC-1α in PPARβ/δ over-expressed muscles at 24h and 54h after termination of exercise were increased 2.5 fold and 2.2 fold, respectively, than Sed group, but PGC-1α ubiquitination was not increased at 24h and 54h after termination of exercise. Our results indicate that PPARβ/δ over-expression does not increase PGC-1α mRNA stability, but increase PGC-1α protein stability through post-translation mechanism after termination of exercise.

• Molecules and Cells
• /
• v.42 no.7
• /
• pp.523-529
• /
• 2019

mRNA quality is controlled by multiple RNA surveillance machineries to reduce errors during gene expression processes in eukaryotic cells. Nonsense-mediated mRNA decay (NMD) is a well-characterized mechanism that degrades error-containing transcripts during translation. The ATP-dependent RNA helicase up-frameshift 1 (UPF1) is a key player in NMD that is mostly prevalent in the cytoplasm. However, recent studies on UPF1-RNA interaction suggest more comprehensive roles of UPF1 on diverse forms of target transcripts. Here we used subcellular fractionation and immunofluorescence to understand such complex functions of UPF1. We demonstrated that UPF1 can be localized to the nucleus and predominantly associated with the chromatin. Moreover, we showed that UPF1 associates more strongly with the chromatin when the transcription elongation and translation inhibitors were used. These findings suggest a novel role of UPF1 in transcription elongation-coupled RNA machinery in the chromatin, as well as in translation-coupled NMD in the cytoplasm. Thus, we propose that cytoplasmic UPF1-centric RNA surveillance mechanism could be extended further up to the chromatin-associated UPF1 and co-transcriptional RNA surveillance. Our findings could provide the mechanistic insights on extensive regulatory roles of UPF1 for many cellular RNAs.

• Korean Journal of Veterinary Research
• /
• v.44 no.2
• /
• pp.163-169
• /
• 2004

To elucidate the molecular mechanisms of autoimmune inflammation in the central nervous system, we examined the expression and localization of STAT1, STAT3, STAT4 and STAT6 molecules during experimental autoimmune encephalomyelitis (EAE) by competitive PCR. In the present study, we quantitated IL-4 and IL-12 p40 mRNA by competitive PCR in the CNS during EAE. IL-4 mRNA was found at early and peak stages. On the other hand, the IL-12 p40 mRNA level reached maximal levels at the peak stage and still found at the recovery stage of the disease. We examined the kinetics of STAT mRNA in the CNS during EAE and demonstrated that STAT1 and STAT4 mRNA reached a maximal level at the peak stage of EAE, whereas STAT3 mRNA level increased gradually to the recovery stage. STAT6 mRNA increased rapidly at the early stage followed by gradual decrease till the recovery stage. Taken together, these findings suggest that STAT4 which was probably activated by IL-12 plays a pro-inflammatory role and that STAT3 which was activated throughout the disease course seems to serve as a transducer of anti-inflammatory signals.

• Toxicological Research
• /
• v.31 no.2
• /
• pp.191-201
• /
• 2015

We evaluated the inhibitory effect of Pueraria lobata root ethanol extract (PLREE) on lipid accumulation during 3T3-L1 differentiation to adipocytes by measuring the intracellular expression of adipogenic, lipogenic, and lipolytic markers and lipid accumulation. The total polyphenol and flavonoid content of PLREE were 47 and 29 mg/g, respectively. The electron donating capacity of PLREE at $1,000{\mu}g/mL$ was 48.8%. Treatment of 3T3-L1 preadipocytes with 100, 250, or $500{\mu}g/mL$ PLREE for 8 days dose-dependently promoted the differentiation of 3T3-L1 cells. In contrast, the lipid content of PLREE-treated cells was significantly reduced by 7.8% (p < 0.05), 35.6% (p < 0.001), and 42.2% (p < 0.001) following treatment with 100, 250, and $500{\mu}g/mL$ PLREE, respectively, as compared to differentiated control cells. PLREE upregulated peroxisome proliferator-activated receptor ${\gamma}$ mRNA and protein, and sterol regulator element-binding protein-1c mRNA levels, but did not affect CCAAT/enhancer binding-protein ${\beta}$ and ${\alpha}$ mRNA levels. PLREE also downregulated acetyl-CoA carboxylase mRNA and protein, fatty acid synthase (FAS) protein, and leptin mRNA levels, but did not affect FAS mRNA expression. PLREE upregulated adipose triglyceride lipase mRNA and protein expression, and hormone-sensitive lipase (HSL) protein expression, but did not affect HSL mRNA expression. In conclusion, we found that PLREE enhanced adipogenesis, but reduced lipogenesis, resulting in decreased lipid accumulation in 3T3-L1 cells.