• Title/Summary/Keyword: E3 protein

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Emodin Inhibits Breast Cancer Cell Proliferation through the ERα-MAPK/Akt-Cyclin D1/Bcl-2 Signaling Pathway

  • Sui, Jia-Qi;Xie, Kun-Peng;Zou, Wei;Xie, Ming-Jie
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.15
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    • pp.6247-6251
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    • 2014
  • Background: The aim of the present study was to investigate the involvement of emodin on the growth of human breast cancer MCF-7 and MDA-MB-231 cells and the estrogen (E2) signal pathway in vitro. Materials and Methods: MTT assays were used to detect the effects of emodin on E2 induced proliferation of MCF-7 and MDA-MB-231 cells. Flow cytometry (FCM) was applied to determine the effect of emodin on E2-induced apoptosis of MCF-7 cells. Western blotting allowed detection of the effects of emodin on the expression of estrogen receptor ${\alpha}$, cyclin D1 and B-cell lymphoma-2 (Bcl-2), mitogen-activated protein kinases (MAPK) and phosphatidylinostiol 3-kinases (PI3K). Luciferase assays were emplyed to assess transcriptional activity of $ER{\alpha}$. Results: Emodin could inhibit E2-induced MCF-7 cell proliferation and anti-apoptosis effects, and arrest the cell cycle in G0/G1 phase, further blocking the effect of E2 on expression and transcriptional activity of $ER{\alpha}$. Moreover, Emodin influenced the ER ${\alpha}$ genomic pathway via downregulation of cyclin D1 and Bcl-2 protein expression, and influenced the non-genomic pathway via decreased PI3K/Akt protein expression. Conclusions: These findings indicate that emodin exerts inhibitory effects on MCF-7 cell proliferation via inhibiting both non-genomic and genomic pathways.

Amoebic PI3K and PKC Is Required for Jurkat T Cell Death Induced by Entamoeba histolytica

  • Lee, Young Ah;Kim, Kyeong Ah;Min, Arim;Shin, Myeong Heon
    • Parasites, Hosts and Diseases
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    • v.52 no.4
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    • pp.355-365
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    • 2014
  • The enteric protozoan parasite Entamoeba histolytica is the causative agent of human amebiasis. During infection, adherence of E. histolytica through Gal/GalNAc lectin on the surface of the amoeba can induce caspase-3-dependent or -independent host cell death. Phosphorylinositol 3-kinase (PI3K) and protein kinase C (PKC) in E. histolytica play an important function in the adhesion, killing, or phagocytosis of target cells. In this study, we examined the role of amoebic PI3K and PKC in amoeba-induced apoptotic cell death in Jurkat T cells. When Jurkat T cells were incubated with E. histolytica trophozoites, phosphatidylserine (PS) externalization and DNA fragmentation in Jurkat cells were markedly increased compared to those of cells incubated with medium alone. However, when amoebae were pretreated with a PI3K inhibitor, wortmannin before being incubated with E. histolytica, E. histolytica-induced PS externalization and DNA fragmentation in Jurkat cells were significantly reduced compared to results for amoebae pretreated with DMSO. In addition, pretreatment of amoebae with a PKC inhibitor, staurosporine strongly inhibited Jurkat T cell death. However, E. histolytica-induced cleavage of caspase-3, -6, and -7 were not inhibited by pretreatment of amoebae with wortmannin or staurosporin. In addition, we found that amoebic PI3K and PKC have an important role on amoeba adhesion to host compartment. These results suggest that amebic PI3K and PKC activation may play an important role in caspase-independent cell death in Entamoeba-induced apoptosis.

Study on Biochemical Pollutant Markers for Diagnosis of Marine Pollution VI. Changes in Cholinesterase Activity of Flounder (Pleuronichthys cornutus) in the Yellow Sea (해양오염의 진단을 위한 생화학적 오염지표에 관한 연구 VI. 황해산 도다리 (Pleuronichthys cornutus)의 콜린에스테라아제 활성의 변화)

  • CHOI Jin-Ho;KIM Dong-Woo;PARK Chung-Kil;YANG Dong Beom
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.4
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    • pp.614-619
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    • 1997
  • This study was designed to investigate the biochemical pollutant markers for diagnosis of marine pollutions by changes in cholinesterase activity of the flounder (Pleuronichthys cornutus) in the Yellow Sea of Korea. Acetylchotinesterase (AChE) activities in brain and muscle of wild flounders in the Yellow Sea were significantly lower $(20\~30\%\;and\;10\~40\%,\;respectively)$ than those of wild flounder in Pohang (control) of the last Sea. Butyrylcholinesterase (BChE) activities in brain and muscle of wild flounders in the Yellow Sea were significantly lower $(10\~30\%\;and\;35\~45\%,\;respectively)$ than those of wild flounder in Pohang of the East Sea. lactate dehydrogenase (LDH) activities in serum of wild flounders in the yellow Sea were significantly $(about\;30\%)$ lower than those of wild flounder in Pohang of the East Sea. These results suggest that AChE and BChE activities in brain and muscle of wild flounders of the Yellow Sea may be used as the most effective mean in a biochemical marker for diagnosis of pollutant effects by organophosphorus pesticides.

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Circadian Clock Gene Per1 Mediates BMP2-induced Osteoblast Differentiation in MC3T3-E1 Cells (MC3T3-E1 세포에서 BMP2에 의한 조골세포의 분화에 일주기 유전자 Per1이 미치는 영향)

  • Min, Hyeon-Young;Jang, Won-Gu
    • Journal of Life Science
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    • v.27 no.5
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    • pp.501-508
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    • 2017
  • Bone morphogenetic proteins (BMPs) are multifunctional cytokines that play important roles in a variety of cellular functions. Among BMP family members, BMP2 efficiently promotes osteoblast differentiation through Smad-mediated runt-related transcription factor 2 (Runx2) expression. Several recent studies suggest that BMPs are associated with clock genes, in particular Bmal1. Bmal1 protein heterodimerizes with Clock protein and then induces period 1 (Per1) expression. However, the role of Per1 on osteoblast differentiation remains unclear. In this study, we investigated whether Per1 is involved in osteoblast differentiation. MC3T3-E1 cells were treated with BMP2 for induction of osteoblastic differentiation. Osteogenic maker gene and Per1 mRNA expression were measured using real-time PCR. Interestingly, BMP2 treatment induced Per1 mRNA expression in MC3T3-E1 cells. To further investigate the function of Per1 on osteoblast differentiation, MC3T3-E1 cells were transiently transfected with pCMV-Per1. Per1 overexpression increased Runx2 mRNA and protein levels. Also, mRNA expression and promoter activity of osteocalcin were upregulated by Per1 overexpression. To investigate the effect of interaction between Per1 and osteogenic condition, MC3T3-E1 cells were cultured in osteogenic medium containing ascorbic acid and ${\beta}$-glycerophosphate. Osteogenic medium-induced ALP staining level and mineralization were synergistically increased by overexpression of Per1. Taken together, these results demonstrate that Per1 is a positive regulator of osteoblast differentiation.

Quality Evaluation of Commercial Extruded Pellet Diet for Olive Flounder, Paralichtys olivaceus (시판 넙치용 배합사료의 품질평가)

  • JI Seung Cheol;MOON Gyeong Su;YOO Jin Huyng;LEE Si Woo;KIM Hong Beom;JEONG Gwan Sik
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.38 no.5
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    • pp.291-297
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    • 2005
  • This study evaluated the quality of commercial extruded pellet (EP) diet of five companies (A, B, C, D and E) for olive flounder Paralichtys olivaceus by biochemical analyses, physical properties and growth performance. The proximate analyses of five EP diets showed $3.2-10.0\%$ of moisture, $49.3-55.5\%$ of crude protein, $4.6-14.7\%$ of crude lipid, $7.0-13.8\%$ of crude ash, $0.7-10.5\%$ of crude fiber, $10.0-27.3\%$ of itrogen free extract (NFE), 304.3-395.4kcal/100g of digestible energy (DE) and 6.1-7.1 of calorie/protein ratio (C/P). Peroxide value (POV) was highest in diet D (47.4 meq/kg) as compared to other diets which in the range of 4.0-11.7 meq/kg. Total amino acid contents were ranged from 46.54 to $55.46\%$ with the highest content in diet B and the lowest content in diet C. Essential amino acid of diet C was lowest $(7.43\%)$ as compared to other diets which in the range of $19.43-20.30\%$. Saturated fatty acid was higher in diet A $(37.65\%)$ followed by diet B $(36.32\%)$, diet E$(34.39\%)$, diet C$(30.95\%)$ and diet D$(30.10\%)$. EPA+DHA were highest in diet E$(30.78\%)$ and lowest in diet C$(15.48\%)$. The floating rate after 6 hours on the sea water was highest in diet C$(100\%)$ followed by diet B$(40\%)$ and A$(10\%)$. However, diets D and E were completely settled down after 1 and 2 hours, respectively. The range of relative expansion rate was $27.2-49.3\%$ for all diets and all reached the peak at 2-3 hours. The water absorption rate of diets C and D was lowest, and diet E was highest at 1 hour after deposition of sea water. Growth rate was higher in diet B$(22.3\%)$ and E$(21.3\%)$. Feed efficiency was higher in diet A$(109.7\%)$ and E$(105.3\%)$ and was significantly lowest in diet D$(80.7\%)$. The protein efficiency ratio was highest in diet E (2.72) and lowest in diet D (1.76). These results suggest that there is a necessity for improvement of nutrients balance and feed physical properties to fulfill the nutrient requirements and digestive characteristics of fishes in commercial EP diets.

Construction of ELISA System for the Detection of Indian citrus ringspot virus (Indian citrus ringspot virus의 ELISA 진단 시스템 구축)

  • Shin, Myeung-Ju;Kwon, Young-Chul;Ro, Hyeon-Su;Lee, Hyun-Sook
    • Research in Plant Disease
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    • v.18 no.3
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    • pp.231-235
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    • 2012
  • Indian citrus ring spot virus (ICRSV) is known to cause a serious disease to citrus, especially to Kinnow mandarin, the popular cultivated citrus species in India. In this study, we developed diagnostic systems based on enzyme-linked immunosorbent assay (ELISA). In order to generate antibodies against ICRSV coat protein, we overexpressed the coat protein in Escherichia coli using the pET15b expression vector containing an optimized ICRSV coat protein gene. The recombinant ICRSV coat protein was overexpressed as soluble form at $37^{\circ}C$ upon IPTG induction. The protein was purified to 95% in purity by Ni-NTA column chromatography. The purified protein was immunized to rabbit for the generation of polyclonal antibody (PAb). The PAb showed a specific immunoreaction to recombinant ICRSV coat protein in western blot analysis and ELISA. Diluted rabbit antisera (10,000 fold) could detect less than 10 ng and 5 ng of the target protein in western blot and ELISA analysis, respectively.

Expression of Human Immunodeficiency Virus Type 1 Gag Protein in Escherichia coli

  • Park, Weon-Sang
    • Journal of Life Science
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    • v.9 no.5
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    • pp.556-563
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    • 1999
  • Presence of antibody to the capsid protein p24 is the main diagnostic criterion, since this reflects reliable antibody response to HIV infection. However, it takes about 6-8 weeks for antibody production after infection and people who are infected but antibodies are not produced yet are classified as seronegative. Therefore, there is a strong need for an improved diagnostic method for better health security. As a first step for developing such an improved diagnostic system, gag protein of human immunodificiency virus type 1 was expressed in E. coli DH5$\alpha$. The gag fragment of HIV-1 (including a portion of p17 and whole p24) was amplified by polymerase chain reaction (PCR) and BamH I/EcoR I sites were created during PCR. The amplified DNA fragment was cleaved with BamH I/EcoR I and was subcloned into the GEX-2T vector which had been digested with BamHI/EcoRI, resulting gene fusion with gst gene of pGEX-2T. The recombinant DNA was transferred into E. coli DH5$\alpha$. The transformed bacteria were grown at 37$^{\circ}C$ for 3h and protein expression was induced with 0.1mM IPTG at $25^{\circ}C$ for 3h. Recombinant gag protein or GST-gag fusion protein was purified with glutathione-sepharose 4B bead and migrated as a single band when analyzed by 10% polyacrylamide gel. These proteins were confirmed by immunoblotting with anti-GST goat sera or Korean AIDS patients sera. The results of this study establish the expression and single step pulification of HIV-1 gag protein which can specifically bind with Korean AIDS patients sera.

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Effects of Dietary Ptecticu tenebrifer Powder and Canned Mixtures on Protein Digestibility by Different Breeds of Companion Dogs (반려견 품종별 동애등에 분말과 통조림 혼합사료 급여가 단백질 소화율에 미치는 영향)

  • Choi, In-Hag;Choi, Sung-Up;Jeong, Yeon-Woo;Park, Kwan-Ho;Kim, Tae-Hoon;Park, Ki-Whan;Chung, Tae-Ho
    • Journal of Environmental Science International
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    • v.31 no.3
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    • pp.285-289
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    • 2022
  • This study investigated the effects of dietary Ptecticu tenebrifer powder and canned mixtures on protein digestibility by different breeds of companion dogs (15 Bichons, 15 Malteses, 15 Chihuahuas and 15 Poodles). The mixtures were divided into Diet A, Diet B, Diet C, Diet D, and Diet E, which were supplied from five farms. Twenty-five grams each was mixed with 100 g of each canned food, and a total of 125 g was measured for each breed of dog. The result of component analysis of the mixtures showed the highest protein contents rather than dry matter, crude ash or crude fat. There were statistical significances (p<0.05) in all mixed feeds fed to bichon, maltese, chihuahua and poodle dog. Overall, protein digestibility by the breeds of dog ranged from 87.44% to 97.18%. Result of breed of dog comparison revealed that Diet E by poodle dog had the highest protein digestibility, and the lowest protein digestibility was observed in Diet C by Maltese. In conclusion, the use of dietary Ptecticu tenebrifer powder and canned mixtures did not only increased protein digestibility by different breeds of dog but also maintained normal manure properties.