Differential gene expression profiling in human cholangiocarcinoma cells treated with Clonorchis sinensis excretory-secretory products

Pak, Jhang Ho,Kim, Dong-Wook,Moon, Ju Hyun,Nam, Joo-Hyun,Kim, Jong-Hyeok,Ju, Jung Won,Kim, Tong-Soo,Seo, Sang-Beom Springer-Verlag 2009 Parasitology research Vol.104 No.5

Proteomic analysis of differentially expressed proteins in human cholangiocarcinoma cells treated with Clonorchis sinensis excretory–secretory products

Pak, Jhang Ho,Moon, Ju Hyun,Hwang, Seung-Jun,Cho, Shin-Hyeong,Seo, Sang-Beom,Kim, Tong-Soo Wiley Subscription Services, Inc., A Wiley Company 2009 Journal of cellular biochemistry Vol.108 No.6

<P>Severe Clonorchis sinensis infection is a significant risk factor for malignant changes in bile ducts and surrounding liver tissues occurring as a result of direct contact with C. sinensis worms and their excretory–secretory products (ESP). However, the intrinsic molecular mechanisms involved in these processes remain obscure. To determine the effects of C. sinensis infection on protein expression in host bile duct epithelium, we examined proteomic profile changes in the human cholangiocarcinoma cell line (HuCCT1) treated with ESP at 24 h. Using a combination of 2-DE, quantitative image and MALDI-TOF MS analysis, we identified 83 proteins that were translationally modulated in response to ESP, among which 49 were up-regulated and 34 down-regulated. These proteins were classified under various biological categories, including metabolism, cell structure and architecture, proteolysis, protein modification, transport, signal transduction, and reactive oxygen species (ROS) detoxification. In particular, ESP induced the expression of redox-regulating proteins, including peroxiredoxins (Prdx 2, 3, and 6) and thioredoxin 1 (Trx 1), possibly via intracellular ROS generation. Application of the proteomic approach to identify ESP response proteins should be a prerequisite before further investigation to clarify the molecular pathways and mechanisms involved in C. sinensis infection of host cells. J. Cell. Biochem. 108: 1376–1388, 2009. © 2009 Wiley-Liss, Inc.</P>

Characterization of Echinostoma cinetorchis endoribonuclease, RNase H

Sung-Bin Lim,Seok Ho Cha,Seung Jegal,Hojong Jun,Seo Hye Park,Bo-Young Jeon,Jhang Ho Pak,Young Yil Bakh,Tong-Soo Kim,Hyeong-Woo Lee 대한기생충학열대의학회 2017 The Korean Journal of Parasitology Vol.55 No.4

Echinostoma cinetorchis is an oriental intestinal fluke causing significant pathological damage to the small intestine. The aim of this study was to determine a full-length cDNA sequence of E. cinetorchis endoribonuclease (RNase H; EcRNH) and to elucidate its molecular biological characters. EcRNH consisted of 308 amino acids and showed low similarity to endoribonucleases of other parasites (<40%). EcRNH had an active site centered on a putative DDEED motif instead of DEDD conserved in other species. A recombinant EcRNH produced as a soluble form in Escherichia coli showed enzymatic activity to cleave the 3-O-P bond of RNA in a DNA-RNA duplex, producing 3-hydroxyl and 5-phosphate. These findings may contribute to develop antisense oligonucleotides which could damage echinostomes and other flukes.

Clonorchis sinensis, an oriental liver fluke, as a human biological agent of cholangiocarcinoma: a brief review

( Tong-soo Kim ),( Jhang Ho Pak ),( Jong-bo Kim ),( Young Yil Bahk ) 생화학분자생물학회(구 한국생화학분자생물학회) 2016 BMB Reports Vol.49 No.11

Parasitic diseases remain an unarguable public health problem worldwide. Liver fluke Clonorchis sinensis is a high risk pathogenic parasitic helminth which is endemic predominantly in Asian countries, including Korea, China, Taiwan, Vietnam, and the far eastern parts of Russia, and is still actively transmitted. According to the earlier 8th National Survey on the Prevalence of Intestinal Parasitic Infections in 2012, C. sinensis was revealed as the parasite with highest prevalence of 1.86% in general population among all parasite species surveyed in Korea. This fluke is now classified under one of the definite Group 1 human biological agents (carcinogens) by International Agency of Research on Cancer (IARC) along with two other parasites, Opisthorchis viverrini and Schistosoma haematobium. C. sinensis infestation is mainly linked to liver and biliary disorders, especially cholangiocarcinoma (CCA). For the purposes of this mini-review, we will only focus on C. sinensis and review pathogenesis and carcinogenesis of clonorchiasis, disease condition by C. sinensis infestation, and association between C. sinensis infestation and CCA. In this presentation, we briefly consider the current scientific status for progression of CCA by heavy C. sinensis infestation from the food-borne trematode and development of CCA. [BMB Reports 2016; 49(11): 590-597]

Toll-Like Receptor-Mediated Free Radical Generation in Clonorchis sinensis Excretory-Secretory Product-Treated Cholangiocarcinoma Cells

Young Yil Bahk,Jhang Ho Pak 대한기생충학열대의학회 2016 The Korean Journal of Parasitology Vol.54 No.5

Clonorchiasis, caused by direct contact with Clonorchis sinensis worms and their excretory-secretory products (ESPs), is associated with chronic inflammation, malignant changes in bile ducts, and even cholangiocarcinogenesis. Our previous report revealed that intracellular free radicals enzymatically generated by C. sinensis ESPs cause NF-κBmediated inflammation in human cholangiocarcinoma cells (HuCCT1). Therefore, the present study was conducted to examine the role of upstream Toll-like receptors (TLRs) on the initial host innate immune responses to infection. We found that treatment of HuCCT1 cells with native ESPs induced changes in TLR mRNA levels in a time-dependent manner, concomitant with the generation of free radicals. ESP-mediated free radical generation was markedly attenuated by preincubation of the cells with TLR1-4-neutralizing antibodies, indicating that at least TLR1 through 4 participate in stimulation of the host innate immune responses. These findings indicate that free radicals triggered by ESPs are critically involved in TLR signal transduction. Continuous signaling by this pathway may function in initiating C. sinensis infection-associated inflammation cascades, a detrimental event leading to progression to more severe hepatobiliary diseases.

Drosophila auraria Complex 의 생화학적 계통

이택준,박장호 한국유전학회 1985 Genes & Genomics Vol.7 No.4

Phylogenic relationships among five sibling species, D. auraria, D. biauraria, D. triauraria, D. quadraria and D. subauraria were investigated by SDS polyacrylamide gel electrophoresis (SDS PAGE) and two-dimensional electrophoresis (TDE). Wager soluble protein patterns of the five species were compared by SDS PAGE. Similarities among D. auraria, D. triauraria, and D. quadraria were found though there was a slight difference in density, while the pattern of D. biauraria was similar to D. subauraria. About 100 protein-spots were detected in TDE slab gel. In the comparison between gels, the genetic distances of five species were calculated according to Aquadro and Avises' (1981) equation. The genetic distance between D. biaruaria and D. subauraria was 0.052 the lowest of all and the next 0.070 between D. triauraria and D. quadraria. In contrast with these, the genetic distance between D. auraria and D. subauraria was 0.213 the highest of all and the next 0.204 between D. auraria and D. biauraria. In consequence, it could be found that D. biauraria was more closely related with D. subauraria and D. auraria was distantly related with D. subauraria. The direction of speciation in these sibling species appeared to be made from D. quadraria to D. auraria and D. triauraria, and from D. triauraria to D. biauraria and D. subauraria, respectively.

정상난소 및 난소상피종양에서의 Peroxiredoxin isoform들의 발현

최은선 ( Eun Sun Choi ),박장호 ( Jhang Ho Pak ),김대연 ( Dae Yeon Kim ),김종혁 ( Jong Hyeok Kim ),김용만 ( Yong Man Kim ),남주현 ( Joo Hyun Nam ),목정은 ( Jung Eun Moc ),김영탁 ( Young Tak Kim ) 대한산부인과학회 2007 Obstetrics & Gynecology Science Vol.50 No.3

목적: Peroxiredoxin (Prx)은 일종의 항산화제로 세포대사과정 중에 생성되는 활성산소 스트레스에 대한 세포 방어기전으로 현재까지 6가지 isoforms이 알려져 있다. 최근의 여러 연구들에서 다양한 종류의 악성종양세포에 Prx가 과발현됨이 보고되었으나 아직까지 난소에서는 많이 연구되지 않아 난소에서의 Prx 발현도를 알아보고자 하였다. 연구대상 및 방법: 본원 산부인과에서 1995년 1월부터 2005년 6월 사이에 난소종양으로 진단된 악성과 양성의 장액성 및 점액성 난소종양 각각 5예씩과 정상 난포성 낭종 5예를 대상으로, 보관된 파라핀 조직에 Prx I-VI를 면역조직화학염색을 시행한 후 광학현미경을 이용하여 핵과 세포질을 구분하여 염색정도를 4단계로 육안적 판독하였다. 결과: 조직에서는 전반적으로 세포질에서 Prx III가 경도 내지 중등도로, Prx VI는 중등도 내지 강도로 염색되었다. Prx IV가 정상군에 비해 점액성 종양상피세포에서 세포질에서 강하게 염색되었고, 악성 종양상피에서는 Prx V가 정상군에 비해 세포질에서 강하게 염색되었고 특징적으로 과립상의 양상을 보였다. Prx VI는 정상에 비해 악성종양에서, 또 양성종양에 비해 악성종양에서 핵에서 그 발현이 증가되어 있었다. 결론: 상피성 난소 종양에서 Prx는 과발현을 보였는데 Prx IV는 점액성 종양 세포질에서, Prx V는 악성 종양 세포질에서, 그리고 Prx VI는 주로 세포질에 존재한다는 이전의 연구결과들과 달리 핵에서 높은 발현을 보였다. Objective: To assess the expression pattern of all six Prxs in normal ovarian tissue and epithelial ovarian tumor cell using immunohistochemical staining. Methods: Patients were retrieved from those who had undertaken operation in Obstetrics and Gynecology of our hospital from January 1995 to June 2005. According to the pathologic result, five patients were allocated randomly in each group of malignant serous, malignant mucinous, benign serous and benign mucinous ovarian tumor. And another five with normal ovarian epithelial cell were included for the comparison. Immunohistochemical staining was performed with Prx I to VI antibodies. Using microscopy, we evaluated the immunoreactivities of nucleus and cytoplasm semiquantitatively by dividing into four categories : -; no immunoactivity present, +; weak, ++; moderate, +++; strong staining. Results: The immunopositivity of Prx III in cytoplasm shows weak to moderate and Prx VI moderate to strong in normal ovarian tissue. In mucinous epithelial ovarian tumor cell, cytoplasmic Prx IV shows stronger activity than in normal epithelial cell or serous tumor cell. In malignant epithelial cell, Prx V shows stronger activity in cytoplasm than normal epithelial cell. It shows characteristically granular pattern. Prx VI shows stronger activity in the nucleus of malignant epithelial cell compared to normal epithelial cell or benign tumor epithelial cell. Conclusion: Normal ovarian tissue showed higher affinity for Prx III and VI. In epithelial ovarian tumor, cytosolic Prx IV in mucinous tumor, cytosolic Prx V and nuclear Prx VI in malignant tumor were overexpressed.

Magnobovatol inhibits smooth muscle cell migration by suppressing PDGF-Rβ phosphorylation and inhibiting matrix metalloproteinase-2 expression

KANG, HYREEN,AHN, DONG HYEON,PAK, JHANG HO,SEO, KYEONG-HWA,BAEK, NAM-IN,JANG, SUNG-WUK D.A. Spandidos 2016 INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE Vol.37 No.5

<P>The migration of vascular smooth muscle cells (VSMCs) may play a crucial role in the pathogenesis of vascular diseases, such as atherosclerosis and post-angioplasty restenosis. Platelet-derived growth factor (PDGF)-BB is a potent mitogen for VSMCs and plays an important role in the intimal accumulation of VSMCs. Magnobovatol, a new neolignan from the fruits of <I>Magnolia obovata</I>, has been shown to have anticancer properties. However, the effects of magnobovatol on VSMCs are unknown. In the present study, we examined the effects of magnobovatol on the PDGF-BB-induced migration of mouse and human VSMCs, as well as the underlying mechanisms. Magnobovatol significantly inhibited the PDGF-BB-induced migration of mouse and human VSMCs without inducing cell death (as shown by MTT assay and wound healing assay). Additionally, we demonstrated that magnobovatol significantly blocked the PDGF-BB-induced phosphorylation of the PDGF receptor (PDGF-R), Akt and extracellular signal-regulated kinase (ERK)1/2 by inhibiting the activation of the PDGF-BB signaling pathway. Moreover, in both mouse and human VSMCs, magnobovatol inhibited PDGF-induced matrix metalloproteinase (MMP)-2 expression at the mRNA and protein level, as well as the proteolytic activity of MMP-2 (as shown by western blot analysis, RT-PCR, gelatin zymography and ELISA). In addition, the sprout outgrowth formation of aortic rings induced by PDGF-BB was inhibited by magnobovatol (as shown by aortic ring assay). Taken together, our findings indicate that magnobovatol inhibits VSMC migration by decreasing MMP-2 expression through PDGF-R and the ERK1/2 and Akt pathways. Our data may improve the understanding of the anti-atherogenic effects of magnobovatol in VSMCs.</P>

Transcriptional regulation of the murine 1-cys peroxiredoxin gene by the B cell-specific activator protein, Pax5

Lee, In-Seon,Choi, Won Ho,Kim, Ji Young,Jeong, Jin-Yong,Kim, Mi-Jung,Nam, Joo-Hyun,Kim, Jong-Hyeok,Seo, Sang-Beom,Pak, Jhang Ho Wiley Subscription Services, Inc., A Wiley Company 2008 Journal of cellular biochemistry Vol.104 No.2

<P>Pax5, a member of the paired box gene family of transcription factors, is a B cell-specific activator protein (BSAP) that plays important roles in controlling the expression of lineage- and differentiation-stage specific genes during B lymphopoiesis. We identified two putative Pax5 binding sites in a 668 bp of the murine 1-cys peroxiredoxin (1-cysPrx) promoter region. These sites were located at positions −278 to −262 and −50 to −34 from the translation start site. Gel mobility shift assays showed that recombinant Pax5 protein bound specifically to the nucleotide regions −56 to −24 (MP1 probe) and −284 to −253 (MP2 probe). Furthermore, endogenous Pax5 protein from B lymphoblast cells (IM-9) formed a DNA-protein complex with MP1 and MP2 probes, indicating that Pax5 binding occurs specifically at these sequences in vivo. Transient transfection studies showed that expression of exogenous Pax5 resulted in dose-dependent increases in 1-cysPrx promoter activity, implying that Pax5 functions as a positive transcription factor for 1-cysPrx expression. Further transient cotransfection studies showed that coexpression of Pax5 and histone acetyltransferases (HATs), such as p300, cAMP-response-element-binding protein (CBP) and p300/CBP associated factor (PCAF) enhanced the Pax5-mediated 1-cysPrx promoter activity. Immunoprecipitation studies indicated that Pax5 directly binds to HATs. Chromatin immunoprecipitation assays showed that the recruitment of Pax5 to the promoter induced histone H3 and H4 hyperacetylation of chromatin. Lipopolysaccharides (LPS) stimulation of murine splenocytes resulted in coordinated expression of Pax5 and 1-cysPrx proteins. These findings suggest that Pax5 may function as a transactivator of 1-cysPrx gene expression. J. Cell. Biochem. 104: 465–476, 2008. © 2008 Wiley-Liss, Inc.</P>

Glaucine inhibits breast cancer cell migration and invasion by inhibiting MMP-9 gene expression through the suppression of NF-κB activation

Kang, Hyereen,Jang, Sung-Wuk,Pak, Jhang Ho,Shim, Sungbo Springer US 2015 MOLECULAR AND CELLULAR BIOCHEMISTRY - Vol.403 No.1

<P> Matrix metalloproteinase-9 (MMP-9) plays a central role in the invasion and metastasis of various types of cancer cells. Here, we demonstrate that glaucine, an alkaloid isolated from the plant<I> Corydalis turtschaninovii</I> tuber (Papaveraceae), can inhibit the migration and invasion of human breast cancer cells. We further show that glaucine significantly blocks phorbol 12-myristate 13-acetate (PMA)-induced MMP-9 expression and activity in a dose-dependent manner. Results from reporter gene and electrophoretic mobility shift assays revealed that glaucine inhibits MMP-9 expression by suppressing activation of the nuclear transcription factor nuclear factor-κB (NF-κB). Moreover, glaucine attenuates PMA-induced IκBα degradation and nuclear translocation of NF-κB. Finally, we also found that glaucine inhibits invasion and MMP-9 expression in the highly metastatic MDA-MB-231 breast cancer cell line. Taken together, our findings indicate that the MMP-9 inhibitory activity of glaucine and its abilities to attenuate IκBα and NF-κB activities may be therapeutically useful as a novel means of controlling breast cancer growth and invasiveness.</P>