• Asian Pacific Journal of Cancer Prevention
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• 제17권12호
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• pp.5303-5307
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• 2016

Objective: Brain tumors cause great mortality and morbidity worldwide, and success rates with surgical treatment remain very low. Several recent studies have focused on introduction of novel effective medical therapeutic approaches. Genistein is a member of the isoflavonoid family which has proved to exert anticancer effects. Here we assessed the effects of genistein on the expression of MMP-2 and VEGF in low and high grade gliomas in vitro. Materials and Methods: High and low grade glioma tumor tissue samples were obtained from a total of 16 patients, washed with PBS, cut into small pieces, digested with collagenase type I and cultured in DMEM containing 10% FBS. When cells reached passage 3, they were exposed to genistein and MMP-2 and VEGF gene transcripts were determined by quantitative real time PCR (qRT-PCR). Results: Expression of MMP-2 demonstrated 580-fold reduction in expression in low grade glioma cells post treatment with genistein compared to untreated cells (P value= 0.05). In cells derived from high grade lesions, expression of MMP-2 was 2-fold lower than in controls (P value> 0.05). Genistein caused a 4.7-fold reduction in VEGF transcript in high grade glioma cells (P value> 0.05) but no effects were evident in low grade glioma cells. Conclusion. Based on the data of the present study, low grade glioma cells appear much more sensitive to genistein and this isoflavone might offer an appropriate therapeutic intervention in these patients. Further investigation of this possibility is clearly warranted.

• The Korean Journal of Physiology and Pharmacology
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• 제23권6호
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• pp.493-499
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• 2019

Macrophage-associated inflammation is crucial for the pathogenesis of diverse diseases including metabolic disorders. Rhodanthpyrone (Rho) is an active component of Gentiana rhodantha, which has been used in traditional Chinese medicine to treat inflammation. Although synthesis procedures of RhoA and RhoB were reported, the biological effects of the specific compounds have never been explored. In this study, the anti-inflammatory activity and mechanisms of action of RhoA and RhoB were studied in lipopolysaccharide (LPS)-stimulated macrophages. Pretreatment with RhoA and RhoB decreased inducible nitric oxide synthase and cyclooxygenase-2 expressions in RAW 264.7 cells and in thioglycollate-elicited mouse peritoneal macrophages. In addition, it downregulated transcript levels of several inflammatory genes in LPS-stimulated RAW 264.7 cells, including inflammatory cytokines/chemokines (Tnfa, Il6, and Ccl2) and inflammatory mediators (Nos2 and Ptgs2). Macrophage chemotaxis was also inhibited by treatment with the compounds. Mechanistic studies revealed that RhoA and RhoB suppressed the nuclear factor $(NF)-{\kappa}B$ pathway, but not the canonical mitogen activated protein kinase pathway, in LPS-stimulated condition. Moreover, the inhibitory effect of RhoA and RhoB on inflammatory gene expressions was attenuated by treatment with an $NF-{\kappa}B$ inhibitor. Our findings suggest that RhoA and RhoB play an anti-inflammatory role at least in part by suppressing the $NF-{\kappa}B$ pathway during macrophage-mediated inflammation.

• Molecules and Cells
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• 제45권6호
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• pp.388-402
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• 2022

Malignant meningiomas often show invasive growth that makes complete tumor resection challenging, and they are more prone to recur after radical resection. Invasive meningioma associated transcript 1 (IMAT1) is a long noncoding RNA located on Homo sapiens chromosome 17 that was identified by our team based on absolute expression differences in invasive and non-invasive meningiomas. Our studies indicated that IMAT1 was highly expressed in invasive meningiomas compared with non-invasive meningiomas. In vitro studies showed that IMAT1 promoted meningioma cell invasion through the inactivation of the Krüppel-like factor 4 (KLF4)/hsa-miR22-3p/Snai1 pathway by acting as a sponge for hsa-miR22-3p, and IMAT1 knockdown effectively restored the tumor suppressive properties of KLF4 by preserving its tumor suppressor pathway. In vivo experiments confirmed that IMAT1 silencing could significantly inhibit the growth of subcutaneous tumors and prolong the survival period of tumor-bearing mice. Our findings demonstrated that the high expression of IMAT1 is the inherent reason for the loss of the tumor suppressive properties of KLF4 during meningioma progression. Therefore, we believe that IMAT1 may be a potential biological marker and treatment target for meningiomas.

• IMMUNE NETWORK
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• 제16권1호
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• pp.61-74
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• 2016

Dendritic cells (DCs) are professional antigen-presenting cells that sample their environment and present antigens to naïve T lymphocytes for the subsequent antigen-specific immune responses. DCs exist in a range of distinct subpopulations including plasmacytoid DCs (pDCs) and classical DCs (cDCs), with the latter consisting of the cDC1 and cDC2 lineages. Although the roles of DC-specific transcription factors across the DC subsets have become understood, the posttranscriptional mechanisms that regulate DC development are yet to be elucidated. MicroRNAs (miRNAs) are pivotal posttranscriptional regulators of gene expression in a myriad of biological processes, but their contribution to the immune system is just beginning to surface. In this study, our in-house probe collection was screened to identify miRNAs possibly involved in DC development and function by targeting the transcripts of relevant mouse transcription factors. Examination of DC subsets from the culture of mouse bone marrow with Flt3 ligand identified high expression of miR-124 which was able to target the transcript of TCF4, a transcription factor critical for the development and homeostasis of pDCs. Further expression profiling of mouse DC subsets isolated from in vitro culture as well as via ex vivo purification demonstrated that miR-124 was outstandingly expressed in CD24⁺ cDC1 cells compared to in pDCs and CD172α⁺ cDC2 cells. These results imply that miR-124 is likely involved in the processes of DC subset development by posttranscriptional regulation of a transcription factor(s).

• 한국발생생물학회지:발생과생식
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• 제2권1호
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• pp.21-27
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• 1998

본 연구에서는 흰쥐 자궁에서 pituitary adenylate cyclase-activating polypeptide (PACAP)과 그 수용체 유전자들이 발현되는가와 각각 어떠한 유형의 transcript들이 발현되는가를 조사하였고, 이를 위해 역전사 중합효소 연쇄반응 (RT-PCR)을 시행하였다. 시상하부와 정소에서 공통적으로 존재함이 알려진 PACAP common exon 부위에 해당되는 primer를 사용하여 PCR을 시행한 결과 자궁을 포함한 모든 조직에서 에상대로 297 bp의 band가 확인되었다. 흰쥐 자궁에서 발현되는 PACAP mRNA에 정소 특이적인 exon 1이 포함되는가를 조사하기 위해 정소 특이적인 primer를 사용하여 PCR을 시행한 결과, 정소에서만 예상대로 586 bp의 band가 검출되었고 자궁을 포함한 다른 조직에서는 발견되지 않았다. 흰쥐의 PACAP 수용체 PCR에서는 자궁에서 923 bp와 839 bp크기의 band를 확인할 수 있었으며, 이는 알려진 PACAP type I수용체의 splicing variant 들 가운데 hip-hop (923bp)과 hip- 또는 nop-type (839bp)의 예상 크기와 일치하였다. 인위적으로 성적인 성숙을 유도한 PMSG 주사모델하에서 자궁내 PACAP transcript 수준은 주사 24 시간 후 실험군에서 증가하여 생식주기상 proestus 시기에 해당되는 주사 48 시간까지 증가하였다가 72 시간후에는 control 보다 낮은 수준을 보였는데, 이는 자궁의 PACAP 유전자발현이 생리적으로 조절됨을 나타내는 것이다. 본 연구는 흰쥐의 자궁에서 PACAP과 그 수용체 isoform들의 유전자가 발현됨을 최초로 보고한 것으로 자궁 자체에서 발현되는 PACAP이 autocrine 또는 paracrine하게 자궁의 생리 및 기능 조절을 담당함을 시사한다.

• 생명과학회지
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• 제18권12호
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• pp.1637-1643
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• 2008

HSP90에 노출된 혈관평활근세포에서 IL-6 transcript가 증가하고, IL-6 단백질의 분비가 증가하며, 또한 IL-6 유전자의 promote가 활성화되었다. HSP90에 의한 IL-6 유전자의 promoter 활성화는 dominant negative 형태의 TLR-4와 MyD88에 의하여 크게 감소되었지만, dominant negative 형태의 TLR-3와 TRIF의 영향을 받지 않았다. 그리고 TLR-4의 이합체화(dimerization)를 저해하는 curcumin은 HSP90에 의한 IL-6의 분비 및 IL-6 유전자 promoter 활성화를 억제하였다. 그리고 IL-6 유전자의 promoter의 NF-${\kappa}B$- 또는 C/EBP-binding sequence에 변이는 HSP90에 의한 IL-6 유전자의 promoter 활성화 억제하였다. 이러한 결과는 혈관평활근세포에서 HSP90에 의한 IL-6 유전자 활성화에 TLR-4와 NF-${\kappa}B$B가 관여함을 의미한다.

• 생명과학회지
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• 제26권7호
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• pp.855-867
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• 2016

말은 인류에 의해 상대적으로 일찍 가축화된 종 중 하나로써, 경주능력, 강건성 및 항병성 등과 같은 능력을 위해 인공적으로 선택되었다. 그 결과, 현재 경주마로 많이 쓰이고 있는 서러브레드의 게놈은 운동 능력에 특화된 유전자형을 많이 갖고 있다. 최근 NGS 기술의 도래와 함께 전장게놈을 대상으로 경주마의 우수한 유전형질을 찾는 연구가 유전체학의 관점에서 진행되고 있다. 그 결과 말의 게놈에 대해서도 GWAS (Genome-wide Association study)가 적용되고 있고, 우수 경주능력을 나타내는 유전자 마커가 발굴되고 있다. 아울러, 특정 샘플의 전장 전사체를 NGS 기법으로 분석할 수 있는 RNA-Seq 기법 역시 활용되고 있는데, 이를 통하여 각 개체별, 운동 전후, 한 개체의 조직별 특정 유전자의 발현 양상과 함께 전사체의 서열 등을 확인할 수 있다. DNA 서열의 변화 없이 유전자 발현을 조절하는 강력한 인자로써 DNA methylation이 주목받고 있다. 말의 게놈에 있어서도 운동 특이적 또는 개체 특이적 DNA methylation 패턴을 보여 주었고, 이는 우수 개체 선정을 위한 마커 개발에 좋은 단서를 제공해 줄 것이다. 유전자 발현을 억제하는 miRNA와, 포유동물의 유전체 내 절반 정도를 차지하고 있는 이동성 유전인자는 기능유전체 연구에 있어서 중요한 인자들이다. 이들은 인간의 게놈에서 많이 연구가 되어 왔으나, 말에서의 연구는 현재 미미한 실정이다. 하지만, 현재까지 말에서 되어 있는 위의 두 인자에 대한 연구 현황을 알아보고, 차후 우수 마 선별 연구에 적용될 가능성을 제시하였다. 기능유전체 및 후성유전체 분석 기법이 발전함에 따라 말에서도 본 연구에서 소개된 여러 가지 분석 기법이 적용되고, 우수한 경주마를 선정하는 데 많은 도움을 줄 것으로 기대하고 있다. 이에 현재까지의 우수한 경주마를 선택하기 위한 많은 연구들 및, 말 연구에 대한 앞으로의 발전 가능성에 대해 고찰하고 토의하였다.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.51-51
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• 2001

Cocaine and amphetamine-regulated transcript (CART) is a satiety factor that is regulated by leptin. It was reported that the mice intracerebroventricularly injected with CART showed behavioral changes resembled with the typical behavioral alterations found in the mice carrying disorders in the brain serotonergic (5-HT) system. Hence, this study was conducted to find out the relationships between CART and 5-HT. We first examined the mRNA levels of CART after the injections of para-chlorophenylalanine (pCPA, 300 mg/kg i.p., single injection or daily for three consecutive days) in the rat brains by in situ hybridization using the mouse CART cDNA probe cloned in our laboratory. Systemic administrations of pCPA, a potent inhibitor of tryptophan hydroxylase, the rate limiting enzyme of 5-HT biosynthesis, acutely depletes the brain 5-HT transporter (5-HTT) in the dorsal raphe nucleus (DRN), which reuptakes terminal 5-HT. Results indicated that the mRNA level of CART significantly decreased in the arcuate nucleus, paraventricular nucleus, and lateral hypothalamic nucleus by three days of daily injection with pCPA with no noticeable change detected 24 hrs after the single injection. The message levels of 5-HTT in DRN decreased in both single and three days of injections. Secondly, to investigate whether CART affect to 5-HT, mouse genomic CART gene, which is consist of 3 exons and 2 introns and mouse neurofilament light (NF-L) chain promoter were cloned. Then, we constructed neuron specific expression vector, which was transfected into HeLa cell using lipid-mediated transfection system. Expression of GFP and CART linked to NF-L-chain promoter in the transfected HeLa cell were detected by using fluorescent microscope and RT-PCR. These results confirmed normal expression of DNA constructs in vitro. Then, to increase brain specific expression of CART in vivo transgenic mice carrying CART gene controlled the deleted NF-L-chain promoter were generated by the DNA microinjection into pronuclei of fertilized embryos. Transgenic mice were detected by Southern blot. Further study is necessary to examine CART expression and 5-HTT in these transgenic mice. Therefore, these results suggest that there maybe a positive molecular correlation between CART and 5-HT in responding to the stimuli.

• 생명과학회지
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• 제25권10호
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• pp.1098-1102
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• 2015

본 연구에서는 C2C12 근육 세포의 분화 과정에 있어 IGF-I이 지방산의 수송을 담당하는 FABPpm mRNA 및 단백질 발현에 어떠한 영향을 미치는지에 대해 알아보았다. 그 결과 근육세포의 분화에 있어 FABPpm의 단백질과 mRNA 발현이 IGF-I에 의해 유의하게 조절되었음을 알 수 있었다. 이는 기존의 여러 연구의 결과인 IGF-I이 골격근에서 근육 관련 유전자들의 발현을 조절하여 근부피 유지 및 증대에 중심적인 역할 것뿐만 아니라, 지방산의 수송을 담당하는 FABPpm의 발현에도 영향을 미친다는 사실을 밝혔다는 것에 의의가 있다고 생각된다. 향후 골격근에서 IGF-I에 의한 FABPpm 발현 조절에 있어, 세포 내부에서의 신호전달 경로 및 상호작용 인자들에 관한 연구를 통해 지방 대사를 조절하는 기전에 관한 연구가 필요할 것으로 사료된다.

• BMB Reports
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• 제40권4호
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• pp.467-474
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• 2007

Autosomal recessive polycystic kidney disease (ARPKD) is one of the important genetic disorders in pediatric practice. Mutation of the polycystic kidney and hepatic disease gene 1 (PKHD1) was identified as the cause of ARPKD. The gene encodes a 67-exon transcript for a large protein of 4074 amino acids termed fibrocystin, but its function remains unknown. The neoplastic-like in cystic epithelial proliferation and the epidermal growth factor/epidermal growth factor receptor (EGF/EGFR) axis overactivity are known as the most important characteristics of ARPKD. Since the misregulation of $Ca^{2+}$ signaling may lead to aberrant structure and function of the collecting ducts in kidney of rat with ARPKD, present study aimed to investigate the further mechanisms of abnormal proliferation of cystic cells by inhibition of PKHD1 expression. For this, a stable PKHD1-silenced HEK-293T cell line was established. Then cell proliferation rates, intracellular $Ca^{2+}$ concentration and extracellular signal-regulated kinase 1/2 (ERK1/2) activity were assessed after treatment with EGF, a calcium channel blocker and agonist, verapamil and Bay K8644. It was found that PKHD1-silenced HEK-293T cell lines were hyperproliferative to EGF stimulation. Also PKHD1-silencing lowered the intracellular $Ca^{2+}$ and caused EGF-induced ERK1/2 overactivation in the cells. An increase of intracellular $Ca^{2+}$ in PKHD1-silenced cells repressed the EGF-dependent ERK1/2 activation and the hyperproliferative response to EGF stimulation. Thus, inhibition of PKHD1 can cause EGF-induced excessive proliferation through decreasing intracellular $Ca^{2+}$ resulting in EGF-induced ERK1/2 activation. Our results suggest that the loss of fibrocystin may lead to abnormal proliferation in kidney epithelial cells and cyst formation in ARPKD by modulation of intracellular $Ca^{2+}$.