• Asian-Australasian Journal of Animal Sciences
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• 제28권11호
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• pp.1532-1536
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• 2015

To verified the target genes of miR-34c, bioinformatics software was used to predict the targets of miR-34c. Three possible target genes of miR-34c related to spermatogenesis and male reproductive development: zinc finger protein 148 (ZNF148), kruppel-like factor 4 (KLF4), and platelet-derived growth factor receptor alpha (PDGFRA) were predicted. Then, the expression of miR-34c and its target genes were detected in swine testicular tissue at different developmental stages by quantitative polymerase chain reaction. The results suggested that the expression of PDGFRA has the highest negative correlation with miR-34c. Then immunohistochemical staining was done to observe the morphology of swine testicular tissue at 2-days and 3, 4, 5-months of age, which indicated that PDGFRA was mainly expressed in the support cells near the basement membrane during the early development stages of testicular tissue, but that the expression of PDGFRA was gradually reduced in later stages. Therefore, western blot analyzed that the highest expression of PDGFRA was generated in 2-days old testicular tissues and the expression levels reduced at 3 and 4-months old, which correlated with the results of immunohistochemical staining. In conclusion, PDGFRA is a target gene of miR-34c.

• Journal of Periodontal and Implant Science
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• 제26권2호
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• pp.396-413
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• 1996

Current acceptable methods for promoting periodontal regeneration are based on removal of diseased soft tissue. root treatment, guided tissue regeneration, introduction of new graft materials and biological mediators. Insulin-like growth factor-I(IGF-I) and Platelet-derived growth factor-BB(PDGF-BB), the members of the polypeptuyde growth factor family have been reported as the biological mediators which regulate a variety cellular matrix biologic activities of wound healing process including the cell proliferation, migration and extracellular matrix synthesis.The purposes of this study is to evaluate the combination effects of IGF-I and PDGF-BB on the cellular activity of the periodontal ligament cells to act as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the first premolar tooth extracted for the orthodontic treatment and were cultured in DMEM containing 10% FBS at the $37^{\circ}C$, 5% CO2 incubator. Author measured the DNA synthetic activity, and total protein, collagen and noncollagenous protein synthetic activities according to the concentration of 10,100ng/ml IGF-I and1,10 ng/ml PDGF-BB in combination. The results were as follows: Significantly increased in the 1 ng/ml PDGF-BB alone compared to the 10 ng/ml PDGF-BB alone(P<0.01) and in the 1 ng/ml PDGF-BB and 10, 100ng/ml IGF-I in combination compared to the 1 ng/ml PDGF-BB alone(P<0.05, P<0.0l). The synthetic activity of the total protein and collagen is significantly increased like to the synthetic activity of the DNA(P<0.05). The synthetic activity of the noncollagenous protein is increased according to the concentration of IGF_I, but not statistically statistically significant(P>0.05). The percent of the collagen is significantly in the 1ng/ml PDGF-BB and 10ng/ml IGF-I in combination compared to the 1ng/ml PDGF-BB alone(P<0.05) and in the 10ng/ml IGF-I in combination compared to the 10ng/ml PDGF-BB alone(P<0.05). The synthetic activity of the DNA is In conclusions, the percent study shows that PDGF-BB and IGF-I in combination have a potentiality to enhance the DNA synthesis and the total protein and collagen synthesis of The periodontal ligament cells, especially it is more significant in the low concentration of PDGF-BB compared to the high one. Thus, the PDGF-BB and IGF-I in combination may have important roles in promotion of periodontal litgment healing, and consequently, may useful for clinical application in periodontal regenerative procedures.

• 대한본초학회지
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• 제29권4호
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• pp.83-89
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• 2014

Objectives : The purpose of this study was to investigate effects of Houttuyniae Herba water extract (HC) on calcium release and production of various inflammatory mediators such as nitric oxide (NO), interferon-inducible protein (IP)-10, platelet derived growth factor (PDGF)-BB, keratinocyte-derived chemokine (KC), vascular endothelial growth factor (VEGF), interleukin (IL)-4, and IL-5 in lipopolysaccharide (LPS)-induced RAW 264.7 mouse macrophages. Methods : NO production was measured by Griess reagent assay. Intracellular calcium level was measured with Fluo-4 assay. Levels of cytokines were measured by High-throughput multiplex bead array cytokine assay based on xMAP (multi-analyte profiling beads) technology. Results : HC significantly decreased NO production for 24 hrs incubation at the concentrations of 10, 25, 50, 100, and $200{\mu}/mL$ in LPS-induced RAW 264.7 (P < 0.05). HC significantly decreased production of IP-10, KC, VEGF, and PDGF-BB for 24 hrs incubation at the concentrations of 50, 100, and $200{\mu}/mL$ in LPS-induced RAW 264.7 (P < 0.05). HC also significantly decreased intracellular calcium release for 24 hrs incubation at the concentrations of 25, 50, 100, and $200{\mu}/mL$ in LPS-induced RAW 264.7 (P < 0.05). But HC did not show any significant effect on production of IL-4 and IL-5 in LPS-induced RAW 264.7. Conclusions : The results suggested that HC has anti-inflammatory property related with its inhibition on the production of NO, IP-10, KC, VEGF, and PDGF-BB in LPS-induced macrophages via calcium pathway.

• Clinical and Experimental Pediatrics
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• 제51권8호
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• pp.879-885
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• 2008

목 적 : 폐섬유아세포는 예전에는 기도의 구조적 세포로만 알려져 왔으나, 최근에는 천식에서 기관지 운동의 톤을 조절할 뿐만 아니라 기도의 면역조절과 기도 개형에서 중요한 역할을 하는 것으로 밝혀지고 있다. VEGF는 혈관 내피세포에서 강력한 작용을 하는 다기능적 사이토카인으로서, 상피내 세포의 세포분열을 유도하고, 상피세포의 투과도를 증가시키며, 상피세포의 이동을 향상시키는 역할을 하는 것으로 알려져 있다. TARC는 Th2 세포의 선택적 이동을 유도하는 케모카인으로 알려져 있다. 본 연구에서는 PDGF와 TGF-${\beta}$로 자극시킨 인간 폐섬유아세포에서 VEGF와 TARC가 생성되는지와 dexamethasone이 폐섬유아세포에서 VEGF의 분비를 억제하는지를 알아보고자 하였다. 또한 폐섬유아세포와 기관지 평활근 세포와 함께 배양했을 때 VEGF 생성에 미치는 효과를 단독배양 시와 비교하였다. 방 법 : 폐섬유아세포와 인간 기관지 평활근세포를 각각 혹은 함께 배양한 뒤 48시간동안 무혈청 배지에서 성장을 정지시킨 후 TGF-${\beta}$ (10 ng/mL)와 PDGF (20 ng/mL)로 자극하였다. 자극 후의 세포 증식 반응과 배양액 상층액의 VEGF, TARC 농도를 측정하여 dexamethasone ($10^{-6}M$)으로 전처치 후 자극한 것과 비교하였다. 결 과 : PDGF와 TGF-${\beta}$로 자극하였을 경우 폐섬유아세포에서 VEGF 분비가 의미있게 증가하였고, 특히 PDGF와 TGF-${\beta}$로 함께 자극하였을 경우 더욱 의미있는 상승을 보였다. Dexamethasone은 폐섬유아세포의 VEGF 분비를 PDGF로 자극한 경우와 PDGF, TGF-${\beta}$ 같이 자극한 경우 모두에서 억제하였다. 인간 기관지 평활근 세포와 폐섬유아세포를 혼합 배양했을 때 VEGF 분비에는 상승적인 효과가 없었다. Dexamethasone은 폐섬유아세포 증식을 억제시키지 않았다. 폐섬유아세포를 PDGF와 TGF-${\beta}$로 자극했을 때 TARC는 분비되지 않았다. 결 론 : 폐섬유아세포는 VEGF 분비를 통해 기도 개형에 관여하며, 기관지 평활근 세포와 함께 배양해도 VEGF 분비에 상승 효과는 없다. Dexamethasone은 VEGF 분비를 억제하였으나 폐섬유아세포의 증식을 억제하지는 못하였다.

• Biomolecules & Therapeutics
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• 제29권5호
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• pp.551-561
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• 2021

Thyroid cancer is the most common endocrine malignancy. Patients with well-differentiated thyroid cancers, such as papillary and follicular cancers, have a favorable prognosis. However, poorly differentiated thyroid cancers, such as medullary, squamous and anaplastic advanced thyroid cancers, are very aggressive and insensitive to radioiodine treatment. Thus, novel therapies that attenuate metastasis are urgently needed. We found that both PDGFC and PDGFRA are predominantly expressed in thyroid cancers and that the survival rate is significantly lower in patients with high PDGFRA expression. This finding indicates the important role of PDGF/PDGFR signaling in thyroid cancer development. Next, we established a SW579 squamous thyroid cancer cell line with 95.6% PDGFRA gene insertion and deletions (indels) through CRISPR/Cas9. Protein and invasion analysis showed a dramatic loss in EMT marker expression and metastatic ability. Furthermore, xenograft tumors derived from PDGFRA geneedited SW579 cells exhibited a minor decrease in tumor growth. However, distant lung metastasis was completely abolished upon PDGFRA gene editing, implying that PDGFRA could be an effective target to inhibit distant metastasis in advanced thyroid cancers. To translate this finding to the clinic, we used the most relevant multikinase inhibitor, imatinib, to inhibit PDGFRA signaling. The results showed that imatinib significantly suppressed cell growth, induced cell cycle arrest and cell death in SW579 cells. Our developed noninvasive apoptosis detection sensor (NIADS) indicated that imatinib induced cell apoptosis through caspase-3 activation. In conclusion, we believe that developing a specific and selective targeted therapy for PDGFRA would effectively suppress PDGFRA-mediated cancer aggressiveness in advanced thyroid cancers.

• Journal of Periodontal and Implant Science
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• 제31권4호
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• pp.787-801
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• 2001

The molecular mechanisms control the function of PDL(periodonta1 ligament) cells and/or fibroblasts remain unclear. PDLsl7, PDL-specific gene, had previousely　identified the cDNA for a novel protein from cultured　PDL fibroblasts using subtraction hybridization between gingival fibroblasts and　PDL fibroblasts. The purpose　of this study was to determine the regulation by growth factors and cytokines on　PDLsl7 gene expression in　cultured human periodontal ligament cells and observe the immunohistochemical　localization of PDLsl7 protein in various tissues of mouse. Primary PDL fibroblasts isolated by scraping the root of the extracted human mandibular third molars. The cells were incubated with various concentration of human recombinant $IL-1{\beta}$, PDGF-BB and TGF\;${\beta}$ for 48h nd 2 weeks. At each time point total RNA was extracted and the levels of transcription ere assessed by reverse transcription-polymerase chain reaction (RT-PCR assay). polyclonal antiserum raised against PDLsl7 peptides, CLSVSYNRSYQINE and SEAVHETDLHDGC, were made, and stained the tooth, periodontium, developing bone, bone marrow and mid-palatal suture of the mouse. The results were as follows. 1. PDLsl7 mRNA levels were increased in response to PDGF (10ng/ml) and $TGF\;{\beta}$(20ng/ml) after treatment of the $IL-1{\beta}$, PDGF-BB and $TGF{\beta}$for 48 h. 2. PDLsl7 was up-regulated only by $TGF{\beta}$(20 ng/ml) after treatment of the $IL-1{\beta}$, PDGF-BB and $TGF\;{\beta}$ for 2 weeks and unchanged by the other stimulants. 3. PDLsl7 was a novel protein coding the 142 amino acid peptides in the ORF and the nucleotide sequences of the obtained cDNA from RT-PCR was exactly same as the nucleotides of the database. 4. Immunohistochemical analysis showed that PDLsl7 is preferentially expressed in the PDL, differentiating osteoblast-like cells and stromal cells of the bone marrow in the adult mouse. 5. The expression of PDLsl7 protein was barely detectable in gingival fibroblasts, hematopoetic cells of the bone marrow and mature osteocytes of the alveolar bone. These results suggest that PDLsl7 might upregulated by PDGF-BB or $TGF{\beta}$ and acts at the initial stage of differentiation when the undifferentiated mesenchymal cells in the bone marrow and PDL differentiate into multiple cell types. However, more research needs to be performed to gain a better understanding of the exact function of PDLsl7 during the differentiation of bone marrow mesenchymal and PDL cells.

• 한국미생물·생명공학회지
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• 제25권6호
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• pp.592-597
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• 1997

During the screening of inhibitors against phospholipase C (PLC) and the formation of inositol phosphates (IP$_{t}$) at NIH3T3${\gamma}$1 cells from microbial secondary metabolites, we selected a fungal strain MT60109 which was capable of producing an inhibitor. By the taxonomic studies, this fungus was identified as Pseudallescheria sp. MT60109 and an inhibitor of PLC was purified by BuOH extraction and chromatographic techniques from the culture broth of Pseudallescheria sp. MT60109. The inhibitor was identified as thielavin B by the physico-chemical properties and spectroscopic analysis of UV, FAB-MS, $^{1}$H, $^{13}$C-NMR, $^{1}$H-$^{1}$H COSY and HMBC. Thielavin B showed potent inhibitory activity against PLC purified from bovine brain with an IC$_{50}$ of 20 $\mu$M. And it also inhibited the formation of inositol phosphates in platelet-derived growth factor (PDGF) -stimulated NIH3T3${\gamma}$1 cells with an IC$_{50}$ of 20 $\mu$M.

• The Korean Journal of Physiology and Pharmacology
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• 제15권4호
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• pp.203-210
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• 2011

Cilostazol is a selective inhibitor of phosphodiesterase 3 that increases intracellular cAMP levels and activates protein kinase A, thereby inhibiting vascular smooth muscle cell (VSMC) proliferation. We investigated whether AMP-activated protein kinase (AMPK) activation induced by heme oxygenase-1 (HO-1) is a mediator of the beneficial effects of cilostazol and whether cilostazol may prevent cell proliferation and reactive oxygen species (ROS) production by activating AMPK in VSMC. In the present study, we investigated VSMC with various concentrations of cilostazol. Treatment with cilostazol increased HO-1 expression and phosphorylation of AMPK in a dose- and time-dependent manner. Cilostazol also significantly decreased platelet-derived growth factor (PDGF)-induced VSMC proliferation and ROS production by activating AMPK induced by HO-1. Pharmacological and genetic inhibition of HO-1 and AMPK blocked the cilostazol-induced inhibition of cell proliferation and ROS production.These data suggest that cilostazol-induced HO-1 expression and AMPK activation might attenuate PDGF-induced VSMC proliferation and ROS production.

• Journal of Periodontal and Implant Science
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• 제25권2호
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• pp.407-417
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• 1995

In order to evaluate the effect of platelet-derived growth factor(PDGF-BB) and guided tissue regeneration(GTR) technique on the regeneration of destructed periodontal tissue,intentional through-and-through furcation defects(4mm in height) were made on both mandibular 2nd and 4th premolars of 8 adult male dogs(30-40lb). Experimental group 1 was composed of the premolars that were treated by only topical application of PDGF-BB with 0.05M acetic acid without any barrier membrane. Experimental group 2 was composed of the premolars that were treated by GTR with expanded polytetrafluoroethylene membrane(ePTFE : Gore-tex periodontal material, USA). Experimental group 3 was composed of the premolars that were treated by GTR with ePTFE after topical application of PDGFBE. Control group was composed of the premolars that were treated by coronally positioned flap operation only without use of PDGF-BB and ePTFE membrane. All ePTFE membranes were carefully removed 4 weeks after regenerative surgery, and all experimental animals were sacrificed 8 weeks after regenerative surgery. The light microscopic findings were as follows ; (1) In experimental group 1, rapid new bone formation along the-root surface with multiple ankylosis and root resorption by multinucleated giant cells, and dense connective tissue in the central portion of the furcation defects were observed. (2) In experimental group 2, it was observed that the furcation defects were filled with newly formed bone, Sharpey's fibers were embedded into new cementum on root dentin of furcation fornix area, but the central portion and the area under furcation fornix were still filled with dense connective tissue. (3) In experimental group 3, the furcation defects were regenerated with newly formed dense bone and regular periodontal ligament with Sharpey's fibers embedded into newly formed cementum and bone underneath fornix area. (4) In control group, unoccupied space, apical migration of epithelium, dense infiltration of inflammatory cells in subepithelial connective tissue in relation to heavy plaque accumulation, and root resorption by inflammatory reaction were shown, but any new cementum formation on resorbed dentin surface could not be observed. The present study demonstrated that the combined therapy of PDGF-BB and GTR could enhance the regeneration of destructed periodontal tissue.

• Journal of Gastric Cancer
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• 제20권1호
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• pp.29-40
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• 2020

Purpose: Gastrointestinal stromal tumors (GISTs) frequently harbor activating gene mutations in either KIT or platelet-derived growth factor receptor A (PDGFRA) and are highly responsive to several selective tyrosine kinase inhibitors. In this study, a targeted next-generation sequencing (NGS) assay with an Oncomine Focus Assay (OFA) panel was used for the genetic characterization of molecular targets in 30 Korean patients with GIST. Materials and Methods: Using the OFA that enables rapid and simultaneous detection of hotspots, single nucleotide variants (SNVs), insertion and deletions (Indels), copy number variants (CNVs), and gene fusions across 52 genes relevant to solid tumors, targeted NGS was performed using genomic DNA extracted from formalin-fixed and paraffin-embedded samples of 30 GISTs. Results: Forty-three hotspot/other likely pathogenic variants (33 SNVs, 8 Indels, and 2 amplifications) in 16 genes were identified in 26 of the 30 GISTs. KIT variants were most frequent (44%, 19/43), followed by 6 variants in PIK3CA, 3 in PDGFRA, 2 each in JAK1 and EGFR, and 1 each in AKT1, ALK, CCND1, CTNNB1, FGFR3, FGFR4, GNA11, GNAQ, JAK3, MET, and SMO. Based on the mutation types, majority of the variants carried missense mutations (60%, 26/43), followed by 8 frameshifts, 6 nonsense, 1 stop-loss, and 2 amplifications. Conclusions: Our study confirmed the advantage of using targeted NGS with a cancer gene panel to efficiently identify mutations associated with GISTs. These findings may provide a molecular genetic basis for developing new drugs targeting these gene mutations for GIST therapy.