• Biomolecules & Therapeutics
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• 제23권2호
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• pp.134-140
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• 2015

Conjugated linoleic acids (CLA) are a family of isomers of linoleic acid. CLA increases growth arrest and apoptosis of human colorectal cancer cells through an isomer-specific manner. ATF3 belongs to the ATF/CREB family of transcription factors and is associated with apoptosis in colorectal cancer. The present study was performed to investigate the molecular mechanism by which t10, c12-CLA stimulates ATF3 expression and apoptosis in human colorectal cancer cells. t10, c12-CLA increased an apoptosis in human colorectal cancer cells in dose dependent manner. t10, c12-CLA induced ATF3 mRNA and luciferase activity of ATF3 promoter in a dose-dependent manner. The responsible region for ATF3 transcriptional activation by t10, c12-CLA is located between -147 and -1850 of ATF3 promoter. mRNA stability of ATF3 was not affected by t10, c12-CLA treatment. t10, c12-CLA increases $GSK3{\beta}$ expression and suppresses IGF-1-stimulated phosphorylation of Akt. The knockdown of ATF3 suppressed expression of $GSK3{\beta}$ and NAG-1 and PARP cleavage. The results suggest that t10, c12-CLA induces apoptosis through ATF3-mediated pathway in human colorectal cancer cells.

• Biomaterials and Biomechanics in Bioengineering
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• 제2권1호
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• pp.33-45
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• 2015

To engineer tissue-like structures, cells are required to organize themselves into three-dimensional networks that mimic the native tissue micro-architecture. Here, we present agarose-based multi-well platform incorporated with electrical stimulation to build skeletal muscle-like tissues in a facile and highly reproducible fashion. Electrical stimulation of C2C12 cells encapsulated in collagen/matrigel hydrogels facilitated the formation 3D muscle tissues. Consequently, we confirmed the transcriptional upregulations of myogenic related genes in the electrical stimulation group compared to non-stimulated control group in our multi-well 3D culture platform. Given the robust fabrication, engineered muscle tissues in multi-well platform may find their use in high-throughput biological studies drug screenings.

• Asian Pacific Journal of Cancer Prevention
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• 제15권8호
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• pp.3471-3476
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• 2014

Background: Aberrant expression of the microRNA-29 family is associated with tumorigenesis and cancer progression. As transport carriers, tumor-derived exosomes are released into the extracellular space and regulate multiple functions of target cells. Thus, we assessed the possibility that exosomes could transport microRNA-29c as a carrier and correlations between microRNA-29c and apoptosis of bladder cancer cells. Materials and Methods: A total of 28 cancer and adjacent tissues were examined by immunohistochemistry to detect BCL-2 and MCL-1 expression. Disease was Ta-T1 in 12 patients, T2-T4 in 16, grade 1 in 8, 2 in 8 and 3 in 12. The expression of microRNA-29c in cancer tissues was detected by quantitative reverse transcriptase PCR (QRT-PCR). An adenovirus containing microRNA-29c was used to infect the BIU-87 human bladder cancer cell line. MicroRNA-29c in exosomes was measured by QRT-PCR. After BIU-87 cells were induced by exosomes-derived microRNA-29c, QRT-PCR was used to detect the level of microRNA-29c. Apoptosis was examined by flow cytometry and BCL-2 and MCL-1 mRNA expressions were assessed by reverse transcription-polymerase chain reaction. Western blotting was used to determine the protein expression of BCL-2 and MCL-1. Results: The expressions of BCL-2 and MCL-1 protein were remarkably increased in bladder carcinoma (p<0.05), but was found mainly in the basal and suprabasal layers in adjacent tissues. The expression of microRNA-29c in cancer tissues was negatively correlated with the BCL-2 and MCL-1. The expression level of microRNA-29c in exosomes and BIU-87 cells from the experiment group was higher than that in control groups (p<0.05). Exosome-derived microRNA-29c induced apoptosis (p<0.01). Although only BCL-2 was reduced at the mRNA level, both BCL-2 and MCL-1 were reduced at the protein level. Conclusions: Human bladder cancer cells infected by microRNA-29c adenovirus can transport microRNA-29c via exosomes. Moreover, exosome-derived microRNA29c induces apoptosis in bladder cancer cells by down-regulating BCL-2 and MCL-1.

• Asian-Australasian Journal of Animal Sciences
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• 제29권12호
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• pp.1790-1795
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• 2016

MicroRNAs (miRNAs) are highly conserved, short non-coding RNAs that regulate gene expression at the posttranscriptional level. Although many miRNAs are identified in muscles and muscle cells, their individual roles are still not fully understood. In the present study, we investigated a muscle highly-expressed miRNA, miR-127-3p, in C2C12 myoblasts and tissues of goats with different muscle phenotypes (Boer vs Wushan black goats). Our results demonstrated that i) miR-127-3p was extensively expressed in tissues of goats; ii) miR-127-3p was higher expressed in muscle, spleen, heart, and skin in the muscular goats (Boer goats) than the control (Wushan black goats). Then we further characterized the dynamical expression of miR-127-3p, MyoD, MyoG, Myf5, Mef2c, and Myosin in the proliferating and differentiating C2C12 myoblasts at day of 0, 1, 3, 5, and 7 in culture mediums. Especially, we found that miR-127-3p was significantly higher expressed in the proliferating than differentiating cells. Our findings suggest that miR-127-3p probably plays roles in the proliferation and differentiation of myoblasts, which further underlies regulation of muscle phenotype in goats.

• 한국축산식품학회지
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• 제41권4호
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• pp.623-635
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• 2021

The effect of deer antler extract on muscle differentiation and muscle atrophy were evaluated to minimize muscle loss following aging. Various deer antler extracts (HWE, hot water extract of deer antler; FE, HWE of fermented deer antler; ET, enzyme-assisted extract of deer antler; UE, extract prepared by ultrasonication of deer antler) were evaluated for their effect on muscle differentiation and inhibition of 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR)-induced muscle atrophy in C2C12 cells. Morphological changes according to the effect of antler extracts on muscle differentiation were confirmed by Jenner-Giemsa staining. In addition, the expression levels of genes related to muscle differentiation and atrophy were confirmed through qRT-PCR. In the presence of antler extracts, the length and thickness of myotubes and myogenin differentiation 1 (MyoD1) and myogenic factor 5 (Myf5) gene expression were increased compared to those in the control group (CON). Gene expression of AMP-activated protein kinase (AMPK), MyoD1, and myogenin, along with the muscle atrophy factors muscle RING finger-1 (MuRF-1) and forkhead box O3a (FoxO3a) upon addition of deer antler extracts to muscle-atrophied C2C12 cells was determined by qRT-PCR after treatment with AICAR. The expression of MuRF-1 and FoxO3a decreased in the groups treated with antler extracts compared to that in the group treated with AICAR alone. In addition, gene expression of MyoD1 and myogenin in the muscle atrophy cell model was significantly increased compared that into the CON. Therefore, our findings indicate that antler extract can increase the expression of MyoD1, Myf5 and myogenin, inhibit muscle atrophy, and promote muscle differentiation.

• 생명과학회지
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• 제28권10호
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• pp.1127-1131
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• 2018

운동 후 분비되는 스트레스 호르몬인 cortisol을 통한 근육세포에 미치는 운동 스트레스의 재현과 coritisol 처리 농도에 따른 세포사멸, 세포질세망 스트레스 및 자가포식현상과의 관련성을 검증 하였다. 마우스 근육 세포주 C2C12를 배양하여 다양한 농도의 cortisol을 12시간 처리하여 세포의 형태 변화를 관찰하고, 세포사멸 마커인 IER3의 발현을 세포면역화학법을 이용하여 확인하였다. 또한 ER-stress와 자가포식 현상의 유도 여부를 확인하기 위하여 BiP와 LC3-I/LC3-II 항체를 이용하여 웨스턴 블랏법을 통해 검증 하였다. 그 결과 cortisol의 농도가 $50{\mu}g/ml$와 $100{\mu}g/ml$로 증가함에 따라 IER3와 BiP 및 LC3-II의 발현량도 유의적으로 증가함을 확인 할 수 있었다. 이러한 결과는 운동 스트레스 호르몬인 cortisol이 운동 후 근육세포의 세포사멸, 세포질세망 스트레스 및 자가포식에 영향을 미침을 보여준다. 본 연구결과는 호르몬과 근육세포 간의 관련성 연구에 기여할 것으로 기대된다.

• Preventive Nutrition and Food Science
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• 제2권3호
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• pp.241-245
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• 1997

Inhibitory effects of kimchi extracts on arcinogen-induced cytotoxicity and transformation in C3H/10T1/2 cells were studied. The methanol extract (500㎍/ml) of fresh (unfermented kimchi), and 3-week-fermented kimchi (properly ripened kimchi at 5℃) inhibited 3-methylcholanthrene (MCA)-induced cytotoxicity in C3H/10T1/2 cells by 84 and 99%, respectively. The inhibitory effect of 3-week-fermented kimchi was higher than that of the fresh kimchi at same test condition. The methanol soluble fraction, and haxane extract of 3-week fermented kimchi also surpressed the cytotoxicity of FC3H/10T1/2 cells mediated by 7,12-dimethylbenz[a]anthracene(DMBA) and N-methyl-N'-nitro-N-nitrosoguanidine(MNNG). Furthermore, MCA-induced transformation of C3H/10T/1/2 cells was significantly inhibited by the methanol soluble fraction of 3-week fermented kimchi. With these results, we suggest that kimchi might have anticarcinogenic effect in part due to inhibition of carcinogen-induced cytotoxicity and transformation of C3H/10T/1/2 cells.

• 대한소아치과학회지
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• 제28권2호
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• pp.238-246
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• 2001

본 실험은 탁월한 골유도능으로 관심의 대상이 되고 있는 BMP의 세포내 신호 전달자로 알려진 Smad 1과 Smad 5가 조골세포 초기 분화표지인자인 ALP 유전자의 발현에 미치는 영향 및 그 조절기전을 알아보고자 하였다. BMP 처리 없이도 Smad에 의해 ALP가 발현되는가를 알아보기 위해 Smad 1과 Smad 5가 각각 stably transfection된 C2C12 세포를 3일간 배양후 histochemical assay를 하였고, Smad 1과 Smad 5의 expression vector와 ALP promoter vector를 transient co-transfection한 후 ALP promoter activity를 측정하였다. Smad에 의한 BMP의 효과를 알아보기 위해서 100ng/ml의 BMP-2를 처리한 군과 처리하지 않은 군으로 나누어 세포를 배양한후 ALP 유전자의 발현을 northern blot analysis로 확인 하였다. Smad가 ALP 유전자의 발현을 직접적으로 조절하는가를 알아보기 위해서는 단백질 합성억제제인 cycloheximide를 전처리하여 ALP 유전자의 발현을 northern blot analysis하였다. 이상의 실험결과 다음과 같은 결론을 얻었다. $\cdot$ Smad 1과 Smad 5가 과발현된 세포에서는 BMP 처리없이도 ALP가 발현된다. $\cdot$ Smad 1과 Smad 5가 과발현된 세포에서 BMP 처리후 ALP 발현 증가율이 대조군 보다 현저히 높게 나타나 Smad가 BMP 효과를 증가시킨다는 것을 알 수 있다. $\cdot$ Smad는 새로운 단백질의 합성을 통해 ALP 유전자를 발현시킨다.

• BMB Reports
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• 제42권10호
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• pp.667-672
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• 2009

Plasma concentrations of adiponectin have been shown to be decreased in patients with obesity, cardiovascular diseases, hypertension and metabolic syndrome. Recent studies have found that adiponectin reduces lipid accumulation in macrophage foam cells which may impact the development of atherosclerosis. However, it remains unclear whether adiponectin is involved in the process of lipid accumulation during myogenesis. Using C2C12 myoblasts, we investigated the effect of adiponectin on intramyocellular lipid accumulation during myogenesis. The results showed that intracellular lipid accumulation is significantly decreased during C2C12 differentiation, apparently due to increased fatty acid oxidation and decreased fatty acid synthesis during this process. C2C12 cells transiently transfected with adiponectin gene showed reduced lipid accumulation as compared to controls. Further experiments demonstrated that adiponectin can suppress lipid accumulation by increasing fatty acid oxidation during C2C12 myogenesis.

• 한국해양학회지:바다
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• 제8권2호
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• pp.78-93
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• 2003

경기만 표영 생태계에서 미소형 및 소형 동물플랑크톤의 군집구조와 생태학적 역할을 조사하기 위하여, 1997년 12월부터 1998년 11월까지 한달 간격으로 미소형 및 소형 동물플랑크톤의 생물량 분포와 군집구조를 조사하였다. 본 조사에서 미소형 및 소형 동물플랑크톤은 2~200 $\mu\textrm{m}$ 크기의 종속영양 미세생물로 정의 하였으며, 종속영양 미소 편모류, 빈 섬모충류와 유종 섬모충류를 포함하는 섬모충류, 종속영양 와편모류와 동물플랑크톤 유생으로 구분하였다. 종속영양 미소 편모류의 현존량과 탄소량은 각각 330~4,370 cells m $l^{-1}$(평균: 1,340$\pm$130 cells m $l^{-1}$), 0.63~12.4 $\mu\textrm{g}$C 1$^{-1}$(평균: 4.35$\pm$0.58 $\mu\textrm{g}$C 1$^{-1}$)로 분포하였고, 부유 섬모충류의 현존량과 탄소량은 각각 338~44,571 cells 1$^{-1}$(평균: 3,526$\pm$544 cells 1$^{-1}$), 1.3~119.7 $\mu\textrm{g}$C 1$^{-1}$(평균: 13.7$\pm$3.0 $\mu\textrm{g}$C 1$^{-1}$)로 분포하였고, 종속영양 와편모류의 현존량과 탄소량은 각각 88~47,461 cells 1$^{-1}$(평균: 9,034$\pm$2,347 cells 1$^{-1}$), 0.04~54.05 $\mu\textrm{g}$C 1$^{-1}$(평균: 6.9$\pm$ 2.4 $\mu\textrm{g}$C 1$^{-1}$)로 분포하였고, 동물플랑크톤 유생의 현존량과 탄소량은 각각 5~546 indiv. 1$^{-1}$(평균: 83$\pm$15 indiv. 1$^{-1}$), 0.17~43.2$\mu\textrm{g}$C 1$^{-1}$(평균: 6.3$\pm$1.2$\mu\textrm{g}$C 1$^{-1}$)로 분포하였다. 조사기간 동안 미소형 및 소형 동물플랑크톤 각 군집의 생물량은 유종 섬모충류를 제외하고는 조석에 의한 차이를 보이지 않았다. 표층부터 저층까지 합산한 미소형 및 소형 동물플랑크톤의 탄소량은 124~l,635 mgC $m^{-}$$^2$/(평균: 585$\pm$110 mgC $m^{-2}$ )로 분포하였으며, 3월과 5월에 가장 높게 나타났다. 미소형 및 소형 동물플랑크톤 중에서 부유 섬모충류가 가장 우점하는 그룹으로 나타났으며, 전체 미소형 및 소형 동물플랑크톤의 평균 42.3%를 기여 하였다. 미소형 및 소형 동물플랑크톤에 대한 각각의 그룹들의 상대적인 기여도는 미소형 및 소형 동물플랑크톤의 군집구조와 계절에 따라 다르게 분포하는 것으로 나타났다. 경기만 표영 생태계서 미소형 및 소형 동물플랑크톤의 군집구조는 식물플랑크톤의 크기별 분포 양상과 유사한 경향을 보였으며, 이 결과는 미소형 및 소형 동물플랑크톤과 식물플랑크톤 간에 피식-포식자의 관계가 있음을 암시하며, 피식-포식의 관계가 미세생물 먹이망에서 중요한 조절 요인임을 보여준다.