• 제목/요약/키워드: ${\beta}$-actin

검색결과 212건 처리시간 0.021초

Production and bioactivity of recombinant tilapia IL-$1\beta$

  • Hong, Su-Hee
    • 한국어병학회지
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    • 제22권2호
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    • pp.147-153
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    • 2009
  • To study the biological activity of interleukin-$1\beta$(IL-$1\beta$), a proinflammatory cytokine, in nile tilapia, Oreochromis niliticus, the recombinant tilapia IL-$1\beta$ was produced in E. coli cells based on pQE vector. Ni-NTA (nitriloacetic acid) metal affinity chromatography was used to purify recombinant protein. The eluted fractions exhibited a single band of protein with a molecular weight of about 25kDa, which is in close agreement with 25.4 kDa predicted by the cDNA sequence. The biological activity of the purified recombinant tilapia IL-$1\beta$ was tested through its effects on IL-$1\beta$ gene expression, which are known as IL-$1\beta$ inducible genes in mammals and fishes. IL-$1\beta$ gene expression induced by poly I:C, a synthetic double stranded RNA, was also assessed in tilapia head kidney cells. IL-$1\beta$ gene expression was analysed using QPCR (quantitative polymerase chain reaction). The ratio of the indicated gene expression was expressed as the relative mRNA level to $\beta$-actin mRNA level, which is constitutively expressed in macrophages. Consequently, head kidney cells incubated for three hours with rIL-$1\beta$(10, 2, 1 $\mu{g}$/ml) showed a dose dependent increase in IL-$1\beta$ mRNA levels and 1 $\mu{g}$/ml of poly I:C was also able to induce IL-$1\beta$ gene expression in head kidney in tilapia.

Profilin-1 과발현에 의한 난소암 세포 성장 및 이동 저해 효능 연구 (Overexpression of Profilin 1 Inhibited Ovarian Tumor Cell Growth and Migration)

  • 이승훈
    • 생명과학회지
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    • 제27권1호
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    • pp.1-7
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    • 2017
  • 비정상적 액틴의 재구성은 암세포의 대표적 특성이다. Thymosin ${\beta}_{10}$ (TB10)과 Profilin-1 (PFN-1)은 액틴중합조절에 필수적인 단백질이다. 이전의 연구에서 본 연구진은 TB10이 F-actin의 구조를 파괴하여 난소 암 세포의 사멸을 일으킨다는 사실을 보고하였으나 그 기전에 대하여 보고 된 바는 아직까지 없다. 본 연구에서는 TB10에 의하여 PFN-1의 발현이 조절되며, PFN-1의 난소 암 저해 유전자로서의 새로운 기능을 보고하였다. 우선 난소암세포주인 SKOV3 세포에서 TB10에 의하여 발현이 조절되는 단백질들을 전기영동법과 liquid chromatography-mass spectroscopy (LC-MS/MS) 방법을 통하여 분석하였다. 그 결과 PFN-1이 TB10에 의하여 발현이 급격히 증가되는 단백질로 동정되었으며, 이 PFN-1을 난소 암 세포주인 SKOV3에 과발현 시켰을 때 암세포의 증식과 이동을 저해하고 암세포 사멸을 유도하였다. 또한 이 결과는 PFN-1에 의하여 Erk 신호전달기전이 저해되고 부수적으로 Elk-1과 Egr-1의 발현이 저해 됨으로써 유도될 가능성을 보여준다. 결론적으로, PFN-1이 난소암세포의 성장과 이동을 저해함과 동시에 세포사멸을 일으키므로 난소 암 치료에 유용하게 이용될 가능성이 높다.

cDNA Microarray를 이용한 치주인대세포와 치은섬유아세포의 유전자 발현에 대한 연구 (A Comparative Study of Gene Expression Patterns of Periodontal Ligament Cells and Gingival Fibroblasts using the cDNA Microarray)

  • 전채영;박진우;이재목;서조영
    • Journal of Periodontal and Implant Science
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    • 제34권1호
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    • pp.205-221
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    • 2004
  • Periodontal ligament(PDL) cells have been known as playing an important roles in periodontal regeneration and gingival fibroblasts are also important to periodontal regeneration by forming connective tissue attachment. There were rare studies about the gene expression patterns of PDL cells and gingival fibroblasts, therefore in this study, we tried cDNA microarray-based gene expression monitoring to explain the functional differences of PDL cells and gingival fibroblasts in vivo and to confirm the characteristics of PDL cells. Total RNA were extracted from PDL cells and gingival fibroblasts of same person and same passages, and mRNA were isolated from the total RNA using Oligotex mRNA midi kit(Qiagen) and then fluorescent cDNA probe were prepared. And microarray hybridization were performed. The gene expression patterns of PDL cells and gingival fibroblasts were quite different. About 400 genes were expressed more highly in the PDL cells than gingival fibroblasts and about 300 genes were more highly expressed in the gingival fibroblasts than PDL cells. Compared growth factor- and growth factor receptor-related gene expression patterns of PDL cells with gingival fibroblasts, IGF-2, IGF-2 associated protein, nerve growth factor, placental bone morphogenic protein, neuron-specific growth- associated protein, FGF receptor, EGF receptor-related gene and PDGF receptor were more highly expressed in the PDL cells than gingival fibroblasts. The results of collagen gene expression patterns showed that collagen type I, type III, type VI and type VII were more highly expressed in the PDL cells than gingival fibroblasts, and in the gingival fibroblasts collagen type V, XII were more highly expressed than PDL cells. The results of osteoblast-related gene expression patterns showed that osteoblast specific cysteine-rich protein were more highly expressed in the PDL cells than gingival fibroblasts. The results of cytoskeletal proteins gene expression patterns showed that a-smooth muscle actin, actin binding protein, smooth muscle myosin heavy chain homolog and myosin light chain were more highly expressed in the PDL cells than gingival fibrobalsts, and ${\beta}-actin$, actin-capping protein(${\beta}$ subunit), actin- related protein Arp3(ARP) and myosin class I(myh-1c) were more highly expressed in the gingival fibroblasts than PDL cells. Osteoprotegerin/osteoclastogenesis inhibitory factor(OPG/OCIF) was more highly expressed in the PDL cells than gingival fibroblasts. According to the results of this study, PDL cells and gingival fibroblasts were quite different gene expression patterns though they are the fibroblast which have similar shape. Therefore PDL cells & gingival fibroblasts are heterogeneous populations which represent distinct characteristics. If more studies about genes that were differently expressed in each PDL cells & gingival fibroblasts would be performed in the future, it would be expected that the characteristics of PDL cells would be more clear.

Bombyx mori β-tubulin Promoter for High-level Expression of Heterologous Genes

  • Park, Kwanho;Goo, Tae-Won
    • International Journal of Industrial Entomology and Biomaterials
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    • 제39권1호
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    • pp.22-28
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    • 2019
  • We previously isolated 9 clones that show stronger signal compared to Bombyx mori cytoplasmic actin gene (BmA3) by using a dot blot hybridization. In this study, we focused on one clone among these clones which has high amino acid similarity with ${\beta}$-tubulin gene of B. mori. This clone was ubiquitously expressed in all tissues and developmental stage of B. mori. As result of promoter assay using dual luciferase assay system, we found the highest transcription activity region (-750/-1) in the 5'-flanking region of ${\beta}$-tubulin gene, which has about 47 fold more intensive promoter activity than BmA3 promoter. Moreover, the ${\beta}$-tubulin promoter was normally regulated in Bm5, Sf9, and S2 cells. Therefore, we suggest that ${\beta}$-tubulin promoter may be used more powerful and effectively for transgene expression in various insects containing B. mori as a universal promoter.

U-937 세포에서 방사선 및 $H_2$O_2$에 의한 ceruloplasmin의 mRNA 유전자 발현 (Ceruloplasmin Gene Expression in U-937 Cells exposed to ${\gamma}$-Irradiation and $H_2$O_2)

  • 오연경;박선영;김인규;윤병수
    • 한국환경성돌연변이발암원학회지
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    • 제22권2호
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    • pp.76-82
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    • 2002
  • In human U-937 cell exposed to ${\gamma}$-irradiation and $H_2O$$_2$, the level of mRNA efrpression in ceruloplasmin gene was measured by using comparative RT.PCR (reverse transcriptase-polymerase chain reaction). At the normal growth condition, the level of ceruloplasmin transcript was estimated as 8.2% and 0.0068% of hprt (hypoxantine phosphoribosyl transferase) transcript and of $\beta$-actin transcript, respectively. In U-937 cells exposed to a dose of 100 rad ${\gamma}$-irradiation, the level of ceruloplasmin transcript was increased about 2.7 and 1.6 fold compared to un-treated cell by using compensation with the levels of hprt and $\beta$-actin transcript. By contrast, the expression of ceruloplasmin gene in U-937 cells exposed to $H_2O$$_2$(50 $\mu$M, 24 h), was shown no significant difference compared to un-treated cell. These results indicated that the expression system of ceruloplasmin gene may react only some specific oxygen species, such as reactive oxygen species induced by ${\gamma}$-irradiation.

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Molecular association of CD98, CD29, and CD147 critically mediates monocytic U937 cell adhesion

  • Kim, Mi-Yeon;Cho, Jae Youl
    • The Korean Journal of Physiology and Pharmacology
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    • 제20권5호
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    • pp.515-523
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    • 2016
  • Adhesion events of monocytes represent an important step in inflammatory responses induced by chemokines. The ${\beta}1$-integrin CD29 is a major adhesion molecule regulating leukocyte migration and extravasation. Although several adhesion molecules have been known as regulators of CD29, the molecular interactions between CD29 and its regulatory adhesion molecules (such as CD98 and CD147) have not been fully elucidated. Therefore, in this study, we examined whether these molecules are functionally, biochemically, and cell-biologically associated using monocytic U937 cells treated with aggregation-stimulating and blocking antibodies, as well as enzyme inhibitors. The surface levels of CD29, CD98, and CD147 (but not CD43, CD44, and CD82) were increased. The activation of CD29, CD98, and CD147 by ligation of them with aggregation-activating antibodies triggered the induction of cell-cell adhesion, and sensitivity to various enzyme inhibitors and aggregation-blocking antibodies was similar for CD29-, CD98-, and CD147-induced U937 cell aggregation. Molecular association between these molecules and the actin cytoskeleton was confirmed by confocal microscopy and immunoprecipitation. These results strongly suggest that CD29 might be modulated by its biochemical and cellular regulators, including CD98 and CD147, via the actin cytoskeleton.

폐쇄성 요로병증에서 $17{\beta}$-estradiol에 의한 신섬유화 감소 효과에 대한 연구 ($17{\beta}$-estradiol Attenuates Renal Fibrosis in Mice with Obstructive Uropathy)

  • 조민현;장희성;정경진;박권무
    • Childhood Kidney Diseases
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    • 제15권2호
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    • pp.125-137
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    • 2011
  • 목적: 일반적으로 남자는 여자에 비해 만성 신장병의 발병이 많고 말기 신부전으로의 진행이 더 흔한 것으로 알려져 있다. 본 연구는 일측성 요관 폐쇄를 가진 생쥐에서 신섬유화에 대한 성별과 성호르몬의 효과를 규명하기 위해 시행되었다. 방법: 일측성 완전 요관 폐쇄 7일째 암컷과 수컷 생쥐의 신장에서 ${\alpha}$-smooth muscle actin (${\alpha}$-SMA)의 발현을 측정한 후, 암컷 생쥐에서 난소를 제거하거나 제거 후 다시 $17{\beta}$-estradiol을 보충하여 나타나는 신섬유화 정도를 비교 분석하였다. 결과:일측성 요관 폐쇄를 가진 암컷 신장의 ${\alpha}$-SMA의 발현이 수컷 신장에 비해 현저히 낮았다. 난소 제거와 $17{\beta}$-estradiol의 보충은 일측성 요관 폐쇄를 가진 암컷 신장의 안지오텐신 II 1형 수용체의 발현에는 의미 있는 영향을 주지 않았지만, 안지오텐신 II 2형 수용체의 발현은 정상 암컷과 난소 제거 후 $17{\beta}$-estradiol를 보충한 암컷에서 현저히 증가되었다. 또한, inducible nitric oxide synthase (iNOS) 역시 유사한 변화를 보였다. 결론 : 여성은 폐쇄성 요로병증에서 신섬유화에 대한 저항성과 연관이 있으며 이러한 성별의 차이는 $17{\beta}$-estradiol에 의한 안지오텐신 II 2형 수용체와 iNOS의 발현 증가와 연관이 있을 것으로 사료된다.

Rat의 DNA Polymerase$\beta$ cDNA가 도입된 Transgenic Drosophila의 체세포 돌연변이 유발에 관한 연구 (Hypersensitivity of Somatic Mutations and Mitotic Recombinations Induced by Mutagens in Transgenic Drosophila bearing Rat DNA Polymerase $\beta$)

  • 최영현;유미애;이원호
    • 한국환경성돌연변이발암원학회지
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    • 제15권2호
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    • pp.100-105
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    • 1995
  • The effects of DNA polymerase $\beta$ on the somatic chromosome mutations and mitotic recombinations were investigated using the transgenic Drosophila beating chimetic gene consisting of a promoter region of Drosophila actin 5C gene and rat DNA polymerase $\beta$. For detecting the somatic chromosome mutations and mitotic recombinations, the heterozygous (mwh/+) strains possessing or lacking transgene poi 13 were used. The spontaneous frequency of small mwh spots, due to deletion or nondisjunction etc., in the non-transgenic w strain and the transgenic p[pol $\beta$]-130 strain was 0.351 and 0.606, respectively. The spontaneous frequency (0.063) of large mwh spots, arises mostly from somatic recombination between the centromere and the locus mwh, in the transgenic p[pol $\beta$]-130 strain was about three times higher than that (0.021) of the non-transgenic w strain. The mutant clone frequencies of small and large mwh spots induced by N-methyl-N'-nitro-N-nitrosoguanidine and ethyl methanesulfonate in the transformant p[pol $\beta$]-130 were higher than those in the host strain w. The present results suggest that rat DNA polymerase $\beta$ participate at least in the somatic chromosome mutations and mitotic recombination processes.

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Red ginseng extract protects against carbon tetrachloride-induced liver fibrosis

  • Ki, Sung Hwan;Yang, Ji Hye;Ku, Sae Kwang;Kim, Sang Chan;Kim, Young Woo;Cho, Il Je
    • Journal of Ginseng Research
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    • 제37권1호
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    • pp.45-53
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    • 2013
  • Korean red ginseng, the processed root of Panax ginseng Meyer, has been frequently used for various therapeutic purposes in oriental medicine. The present study investigated the possible effect of Korean red ginseng extract (RGE) for the treatment of liver fibrosis in mice injected with carbon tetrachloride ($CCl_4$) for 4 wk. Liver injuries were assessed by blood biochemistry and histopathology in mice treated with $CCl_4$ alone or $CCl_4$+ RGE (30, 100, and 300 mg/kg). Concomitant treatment with RGE and $CCl_4$ (three times/wk for 4 wk) effectively inhibited liver fibrosis as evidenced by decreases in plasma alanine and aspartate aminotransferases, as well as by the percentages of degenerative regions, numbers of degenerative hepatocytes, and collagen accumulation in hepatic parenchyma. Treatment with $CCl_4$ for 4 wk increased mRNA levels of transforming growth factor ${\beta}1$ and plasminogen activator inhibitor 1 in fibrogenic liver, whereas RGE (30, 100, and 300 mg/kg) significantly blocked the induction of fibrogenic genes by $CCl_4$. Similarly, RGE also prevented transforming growth factor ${\beta}1$-mediated induction of fibrogenic genes in human hepatic stellate cell lines. More importantly, RGE markedly reduced the number of ${\alpha}$-smooth muscle actin-positive cells in liver tissue. This study implies that RGE efficaciously protects against the liver fibrosis induced by chronic $CCl_4$ treatment, and may therefore have potential to treat liver disease.