Hepatic differentiation of the human hepatic progenitor cells (HPCs) by sodium butyrate (초)

( Pei Pei Hao ),( Xiang Dan Cui ),( Mi Jin Lee ),( Gyung Ran Yu ),( Dae Ghon Kim ) 대한간학회 2010 Clinical and Molecular Hepatology(대한간학회지) Vol.16 No.3(S)

Utilizing Creative Quality Management to Enhance Hand Hygiene among Healthcare Personnel

Pei-Yu Lee,Ju-Chen Yen,Ching-Chi Wang 한국간호과학회 2023 한국간호과학회 학술대회 Vol.2023 No.11

Basic, HCC basic : PE-108 ; Progenitor cell-derived hepatocytes and their characteristics in human

( Pei Pei Hao ),( Mi Jin Lee ),( Goung Ran Yu ),( In Hee Kim ),( Dae Ghon Kim ) 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.-

Background: Hepatic progenitor cells (HPCs) are capable of differentiating along the hepatic lineage into hepatocytes or cholangiocytes (bile duct cells), hence play a critical role in the process of liver regeneration. Their biological discrimination and characterization are critical for therapeutic potential. Aims of this study is to establish progenitor cell-derived hepatocytes and to characterize their specific markers. Methods: Potential liver progenitor cells (HNK-1) were established and their various HPC protein expressions were investigated by immunoblotting, immunofluorescence and fluorescence-activated cell sorting (FACS) analyses, compared with those of other HCC cells. Immunohistochemistry was performed to detect these HPC antigen expression in the tissues of hepatic cirrhosis. Anchorage-independent growth and tumorigenicity were determined using soft agar and xenograft assay. Results: The HNK-1 cells highly expressed HPC markers such as EpCAM, CK7, CK19, AFP, CK8, CK18, EFNA1, and Thy1. Whereas, CD133 was barely expressed. In contrast, malignant Hep3B cells were positive in both EpCAM and CD133. Ductular reactions at the periphery of the cirrhotic nodules were immunohistochemically positive for these HPC markers. Sodium butyrate could induce hepatocyte-like morphological changes in HNK-1 cells, accompanying down-regulation of the hepatic progenitor cell markers (EpCAM, CK7, CK19, and EFNA1) and up-regulation of mature hepatocyte markers (albumin, CK8, and CK18) in both dose-dependent and time- dependent manners. Colony formation in vitro and tumorigenesis in vivo showed that there were no tumorigenesis capacity in EpCAM (+)/CD133(-) HNK1cells at the 0-2nd, 10th, 25th, and 50th passages, while the positive control EpCAM (+)/CD133(+) Hep3B cells could induce tumor in the mice model. Conclusions: Taken together, our results suggest that HNK1 cells are progenitor cell-derived hepatocytes and their stemmness- related markers EpCAM (+)/CD133(-) may be a distinguished marker for nonmalignant, progenitor cell-derived hepatocytes.

Basic, Research : Isolation of EpCAM+/CD133? Hepatic Progenitor Cells

( Pei Pei Hao ),( Mi Jin Lee ),( Goung Ran Yu ),( In Hee Kim ),( Dae Ghon Kim ) 대한간학회 2013 춘·추계 학술대회 (KASL) Vol.2013 No.1

Background/Aim: Hepatic progenitor cells (HPCs) are capable of differentiating along the hepatic lineage into hepatocytes or cholangiocytes. Progenitor cell-derived hepatocytes are critical for hepatocyte replenishment. Therefore, we have established human hepatic progenitor cells (HNK1) and determined their biological characteristics for experimental and therapeutic applications. Methods: Potential liver progenitor cells (HNK1) were established and their various HPC protein expressions were investigated by immunoblotting, immunofluorescence and fluorescence- activated cell sorting (FACS) analyses, compared with those of other HCC cells. Immunohistochemistry was performed to detect these HPC antigen expression in the tissues of hepatic cirrhosis. Albumin, ureagenesis and CYP450 activity were measured. Anchorage-independent growth and tumorigenicity were determined using soft agar and xenograft assay. Genetic constitution of the HNK1 was examined by karyotyping. Chromaosomal rearrangements at metaphase were detedted by Giemsa banding. Results: The HNK1 cells highly expressed HPC markers such as EpCAM, CK7, CK19, AFP, CK8, CK18, EFNA1, and Thy1. Whereas, CD133 was barely expressed. In contrast, malignant Hep3B cells were positive in both EpCAM and CD133. Ductular reactions at the periphery of the cirrhotic nodules were immunohistochemically positive for these HPC markers. Sodium butyrate could induce hepatocyte-like morphological changes in HNK1 cells, accompanying down-regulation of the hepatic progenitor cell markers (EpCAM, CK7, CK19, and EFNA1) and up-regulation of mature hepatocyte markers (albumin, CK8, and CK18). Albumin, ureagenesis, and CYP450 activity were also significantly increased by serial passages after treatment with sodium butyrate. Colony formation in vitro and tumorigenesis in vivo showed that there were no tumorigenesis capacity in EpCAM (+)/CD133(-) HNK1 cells at the 0-2nd,10th,25th,and 50th passages, while the positive co ntrol EpCAM (+)/CD133(+) Hep3B cells could induce tumor in the mice model. Conclusions: HNK1 cells were found to be EpCAM+/CD133? hepatic progenitor cells without spontaneous malignant transformation ability that could be useful for experimental and therapeutic applications. Moreover, EFNA1 should be recognized as an HPC marker.

Basic, HCCbasic : PO-20 ; DUSP1 induces p53 target gene expression through p38MAPK/HSP27 pathway and tumor suppression in hepatocellular carcinoma

( Pei Pei Hao ),( Mi Jin Lee ),( Yun Peng Wang ),( Goung Ran Yu ),( In Hee Kim ),( Dae Ghon Kim ) 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.1

Background: Constitutive DUSP1 expression has been shown to be involved in cell cycle inhibition, apoptosis, and senescence. This study was aimed to examine whether DUSP1 functions as tumor suppressor in hepatocarcinogenesis and to explore underlying mechanism whereby DUSP1 suppresses hepatocarcinogenesis. Methods: Immunohistochemistry and real-time PCR analysis were performed in HCC tissues. Cellular localization of DUSP1 was detected by immunofluorescence. Cell proliferation was tested by MTT assay. Cell death and cell cycle were measured by FACS analysis. Apoptotic and kinase signaling were explored by western blot analysis. Tumorigenicity and survival analysis were tested by xenotransplant of SH-J1 cells stably expressing DUSP1 or infected with Ad-DUSP1 in mouse model. Phospho-related factors expression profile in DUSP1 stable cell lines as determined by phospho-kinase array. Results: The mRNA and protein expression level of DUSP1 was down-regulated in tumor than that of the corresponding non-tumor in HCC tissues. Cellular localization of DUSP1 showed that the endogenous DUSP1 and ectopic expression of GFP-tagged DUSP1 was mainly located in the nucleus. DUSP1 down-regulation was associated with reciprocal activation of ERK1/2 in HCC cell lines. DUSP1 was up-regulated in a dose dependent manner after parthenolide or doxorubicin treatment. DUSP1 over-expression was correlated with the increased susceptibility to apoptotic cell death through caspase activation. Ectopic DUSP1 over-expression resulted in the inhibitions of cell cycle progression, colony generation, and tumor growth in vitro and in vivo system. Furthermore, survival rate of mice xenoplanted with DUSP1 overexpressed HCC cells is significantly higher than control group. Inhibition of tumorigenic potential by DUSP1 may involve in p38MAPK- HSP27-P53 pathway. Conclusions: DUSP1 functions as a tumor suppressor during hepatocarcinogenesis, which seemed to be mainly associated with the activation of p53 target genes through p38MAPK/ HSP27 pathway.

HCV : PE-108 ; Progenitor cell-derived hepatocytes and their characteristics in human

( Pei Pei Hao ),( Mi Jin Lee ),( Goung Ran Yu ),( N Hee Kim ),( Dae Ghon Kim ) 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.1

Background: Hepatic progenitor cells (HPCs) are capable of differentiating along the hepatic lineage into hepatocytes or cholangiocytes (bile duct cells), hence play a critical role in the process of liver regeneration. Their biological discrimination and characterization are critical for therapeutic potential. Aims of this study is to establish progenitor cell-derived hepatocytes and to characterize their specific markers. Methods: Potential liver progenitor cells (HNK-1) were established and their various HPC protein expressions were investigated by immunoblotting, immunofluorescence and fluorescence-activated cell sorting (FACS) analyses, compared with those of other HCC cells. Immunohistochemistry was performed to detect these HPC antigen expression in the tissues of hepatic cirrhosis. Anchorage-independent growth and tumorigenicity were determined using soft agar and xenograft assay. Results: The HNK-1 cells highly expressed HPC markers such as EpCAM, CK7, CK19, AFP, CK8, CK18, EFNA1, and Thy1. Whereas, CD133 was barely expressed. In contrast, malignant Hep3B cells were positive in both EpCAM and CD133. Ductular reactions at the periphery of the cirrhotic nodules were immunohistochemically positive for these HPC markers. Sodium butyrate could induce hepatocyte-like morphological changes in HNK-1 cells, accompanying down-regulation of the hepatic progenitor cell markers (EpCAM, CK7, CK19, and EFNA1) and up-regulation of mature hepatocyte markers (albumin, CK8, and CK18) in both dose-dependent and timedependent manners. Colony formation in vitro and tumorigenesis in vivo showed that there were no tumorigenesis capacity in EpCAM (+)/CD133(-) HNK1cells at the 0-2nd, 10th, 25th, and 50th passages, while the positive control EpCAM (+)/CD133(+) Hep3B cells could induce tumor in the mice model. Conclusions: Taken together, our results suggest that HNK1 cells are progenitor cell-derived hepatocytes and their stemmnessrelated markers EpCAM (+)/CD133(-) may be a distinguished marker for nonmalignant, progenitor cell-derived hepatocytes.

Basic, HCC basic : O-034 ; Pro-oncogenic NM23-H2 modulates MDM2 expression in hepatocarcinogenesis

( Mi Jin Lee ),( Goung Ran Yu ),( Hua Lee ),( Sang Wook Kim ),( Pei Pei Hao ),( In Hee Kim ),( Dae Ghon Kim ) 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.1

Background: NM23 is a family of structurally and functionally conserved proteins known as nucleoside diphosphate kinases (NDPK). NM23-H1 and NM23-H2 are expressed abundantly in HCC. NM23-H2 is a basic protein recently identified as the human PuF factor, which is a transcriptional activator of the c-Myc proto-oncogene. Although the NM23-H1 protein is implicated as a metastasis suppressor, the role of NM23-H2 appears to be less understood. Thus, the aim of this study is to examine functional role and mechanism of NM23-H2 involved tumorigenesis in HCC. Methods: We examined the NM23-H1, H2 and LV mRNA expression in HCC by Realtime-PCR analysis and NM23-H2 protein expression in HCC by immunoblot and immunohistochemistry. Focus formation and anchorage-independent growth were examined in stable cell lines expressing NM23-H2 using soft agar. Using overexpression of NM23-H2 by adenoviral system, the molecular mechanism of NM23-H2 mediated tumor cell growth was assessed in experimental cell culture and in vivo animal model. Results: The level of NM23-H2 expression in tumor tissues and the surrounding matrix appeared to be independent of etiology and tumor differentiation. Ectopic expression of NM23-H2 in NIH3T3 fibroblasts and HLK3 hepatocytes enhanced focus formation, and allowed anchorage-independent growth. Ectopic expression of NM23-H2 induced MDM2 expression. However, MDM2 mRNA and promoter activity was not changed by ectopic expression of NM23-H2, but NM-23H2 interacted with MDM2. The NIH3T3 fibroblasts and HLK3 hepatocytes stably expressing NM23-H2 produced tumors in athymic mice. Lentiviral delivery of NM23-H2 shRNA inhibited tumor growth of xenotransplanted tumors produced from HLK3 cells stably expressing NM23-H2. Conclusions: These results indicate that NM23-H2 may be pro-oncogenic and regulate MDM2 expression in hepatocarcinogenesis. Therefore, this pathway may be an useful target for HCC treatment.

Basic, HCC basic : O-034 ; Pro-oncogenic NM23-H2 modulates MDM2 expression in hepatocarcinogenesis

( Mi Jin Lee ),( Goung Ran Yu ),( Hua Lee ),( Sang Wook Kim ),( Pei Pei Hao ),( In Hee Kim ),( Dae Ghon Kim ) 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.-

Background: NM23 is a family of structurally and functionally conserved proteins known as nucleoside diphosphate kinases (NDPK). NM23-H1 and NM23-H2 are expressed abundantly in HCC. NM23-H2 is a basic protein recently identified as the human PuF factor, which is a transcriptional activator of the c-Myc proto-oncogene. Although the NM23-H1 protein is implicated as a metastasis suppressor, the role of NM23-H2 appears to be less understood. Thus, the aim of this study is to examine functional role and mechanism of NM23-H2 involved tumorigenesis in HCC. Methods: We examined the NM23-H1, H2 and LV mRNA expression in HCC by Realtime-PCR analysis and NM23-H2 protein expression in HCC by immunoblot and immunohistochemistry. Focus formation and anchorage-independent growth were examined in stable cell lines expressing NM23-H2 using soft agar. Using overexpression of NM23-H2 by adenoviral system, the molecular mechanism of NM23-H2 mediated tumor cell growth was assessed in experimental cell culture and in vivo animal model. Results: The level of NM23-H2 expression in tumor tissues and the surrounding matrix appeared to be independent of etiology and tumor differentiation. Ectopic expression of NM23-H2 in NIH3T3 fibroblasts and HLK3 hepatocytes enhanced focus formation, and allowed anchorage-independent growth. Ectopic expression of NM23-H2 induced MDM2 expression. However, MDM2 mRNA and promoter activity was not changed by ectopic expression of NM23-H2, but NM-23H2 interacted with MDM2. The NIH3T3 fibroblasts and HLK3 hepatocytes stably expressing NM23-H2 produced tumors in athymic mice. Lentiviral delivery of NM23-H2 shRNA inhibited tumor growth of xenotransplanted tumors produced from HLK3 cells stably expressing NM23-H2. Conclusions: These results indicate that NM23-H2 may be pro-oncogenic and regulate MDM2 expression in hepatocarcinogenesis. Therefore, this pathway may be an useful target for HCC treatment.

Suppression of Adipogenesis by 5-hydroxy-3,6,7,8,3′,4′-Hexamethoxyflavone from Orange Peel in 3T3-L1 Cells

Yu Wang,Pei-Sheng Lee,Yi-Fen Chen,Chi-Tang Ho,Min-Hsiung Pan 한국식품영양과학회 2016 Journal of medicinal food Vol.19 No.9

We reported previously that hydroxylated polymethoxyflavones (HPMFs) effectively suppressed obesity in high-fat-induced mouse. In this study, we further investigated the molecular mechanism of action of 5-hydroxy-3,6,7,8,3′,4′-hexamethoxyflavone (5-OH-HxMF), one of major HPMFs in orange peel. Treatment of 5-OH-HxMF effectively inhibited lipid accumulation by 55–60% in a dose-dependent manner. The 5-OH-HxMF attenuated adipogenesis through downregulating adipogenesis-related transcription factors such as peroxisome proliferator-activated receptor gamma (PPARγ) and CCAAT/enhancer-binding proteins (C/EBPs), as well as downstream target fatty acid synthase and acetyl-CoA carboxylase (ACC). 5-OH-HxMF activated adenosine monophosphate-activated protein kinase signaling and silent mating type information regulation 1 (SIRTUIN 1 or SIRT1) in 3T3-L1 adipocytes to decrease lipid accumulation. In addition, the inhibition rate of lipid accumulation was compared between 5-OH-HxMF and 3,5,6,7,8,3′,4′-heptamethoxyflavone (HpMF). 5-OH-HxMF inhibited lipid accumulation 15–20% more than HpMF did, indicating that hydroxyl group at position 5 can be a key factor in the suppression of adipogenesis.

Precursor mass prediction by clustering ionization products in LC-MS-based metabolomics

Lee, Terk Shuen,Ho, Ying Swan,Yeo, Hock Chuan,Lin, Joyce Pei Yu,Lee, Dong-Yup Springer-Verlag 2013 Metabolomics Vol.9 No.6