• Biomolecules & Therapeutics
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• v.32 no.1
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• pp.94-103
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• 2024

Non-alcoholic fatty liver disease (NAFLD) is characterized by excessive accumulation of fat in the liver, and there is a global increase in its incidence owing to changes in lifestyle and diet. Recent findings suggest that p53 is involved in the development of non-alcoholic fatty liver disease; however, the association between p53 expression and the disease remains unclear. Doxorubicin, an anticancer agent, increases the expression of p53. Therefore, this study aimed to investigate the role of doxorubicin-induced p53 upregulation in free fatty acid (FFA)-induced intracellular lipid accumulation. HepG2 cells were pretreated with 0.5 ㎍/mL of doxorubicin for 12 h, followed by treatment with FFA (0.5 mM) for 24 h to induce steatosis. Doxorubicin pretreatment upregulated p53 expression and downregulated the expression of endoplasmic reticulum stress- and lipid synthesis-associated genes in the FFA -treated HepG2 cells. Additionally, doxorubicin treatment upregulated the expression of AMP-activated protein kinase, a key modulator of lipid metabolism. Notably, siRNA-targeted p53 knockdown reversed the effects of doxorubicin in HepG2 cells. Moreover, doxorubicin treatment suppressed FFA -induced lipid accumulation in HepG2 spheroids. Conclusively, these results suggest that doxorubicin possesses potential application for the regulation of lipid metabolism by enhance the expression of p53 an in vitro NAFLD model.

• IMMUNE NETWORK
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• v.21 no.5
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• pp.37.1-37.17
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• 2021

Hepatitis B virus X (HBx) protein has been reported as a key protein regulating the pathogenesis of HBV-induced hepatocellular carcinoma (HCC). Recent evidence has shown that HBx is implicated in the activation of autophagy in hepatic cells. Nevertheless, the precise molecular and cellular mechanism by which HBx induces autophagy is still controversial. Herein, we investigated the molecular and cellular mechanism by which HBx is involved in the TRAF6-BECN1-Bcl-2 signaling for the regulation of autophagy in response to TLR4 stimulation, therefore influencing the HCC progression. HBx interacts with BECN1 (Beclin 1) and inhibits the association of the BECN1-Bcl-2 complex, which is known to prevent the assembly of the pre-autophagosomal structure. Furthermore, HBx enhances the interaction between VPS34 and TRAF6-BECN1 complex, increases the ubiquitination of BECN1, and subsequently enhances autophagy induction in response to LPS stimulation. To verify the functional role of HBx in liver cancer progression, we utilized different HCC cell lines, HepG2, SK-Hep-1, and SNU-761. HBx-expressing HepG2 cells exhibited enhanced cell migration, invasion, and cell mobility in response to LPS stimulation compared to those of control HepG2 cells. These results were consistently observed in HBx-expressed SK-Hep-1 and HBx-expressed SNU-761 cells. Taken together, our findings suggest that HBx positively regulates the induction of autophagy through the inhibition of the BECN1-Bcl-2 complex and enhancement of the TRAF6-BECN1-VPS34 complex, leading to enhance liver cancer migration and invasion.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.1
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• pp.114-122
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• 1995

Defining the mechanisms of B cell diversification which establish the immune repertoire is fundamental to understand how the immune response is regulated. In this report, B cell differentiation and diversification focused on the regulation of immunoglobulin $V_H$ gene expression during ontogeny were analyzed by in situ hybridization technique. Fetal liver B cells in .different gestational days from 16d to 20d showed the predominant expression of $V_H7183$ and $V_HQ52$ without transition of repertoire during the observed gestation days. The two subsets of fetal liver B cells separated according to different differentiation stages based on the presence of tell surface immunoglobulin also did not indicate apparent difference in expressed $V_H$ gene family profiles. B cells in fetal spleen as an another hematopoietic lymphoid tissue in fetus also expressed similar $V_H$ gene repertoire to that in fetal liver B cells. This distinct pattern of $V_H$ gene expression in fetal B cells from that of adult B cells were not changed even after four weeks contact with adult bone marrow microenvironment supplied by the established adult bone marrow stromal cell layers. Thus, the restricted $V_H$ gene repertoire of B cells in fetus which is distinct from that in adult appears to be associated more with the genetic potential of fetal B cell progenitors and less with environmental influences or differentiation stages or compartmentalization.

• The Journal of Internal Korean Medicine
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• v.21 no.1
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• pp.65-73
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• 2000

Objectives : The effects of cotreatment of adriamycin and ethanol extract of herb (Palgin-tang hab Hwajuck-hwan a traditional medicine for cancer treatment in oriental medicine) on the induction of apoptotic cell death were investigated in human liver origin cell lines, Chang. Methods : Chang(ATCC) liver cells were cultured in RPMI-1640(Gibco SRL Co, Gaithersburg, MD) badge including 10% fetal bovine serum. Chang liver cells were treated with various concentrations(from 10 to $0.16{\mu}l$) of adriamycin and herb extract(from 500 to $31.25{\mu}l$) After 48h later, the cells were tested for viability by Crystal violet staining assay. Adriamycin and Herb extract induced ladder pattern of DNA fragmentation in Chang cells. Genomic DNA was isolated and separated on 1.5% agarose gels. The DNA was stained with ethidium bromide and visualized under UV light. Results : The death of Chang cells was synergistically induced by the cotreatment of adriamycin and ethanol extract of herb. In addition, the cotreatment-induced cell death of Chang cells was mediated by apoptotic death signal processes. The phosphotransferase activity of JNK1 remained in a basal level in Chang cells which was treated individually with the adriamycin and ethanol extract of herb. However, it was markedly increased in Chang cells which was cotreated with adriamycin and ethanol extract of herb. In addition, the expression of Fas and FasL was markedly induced by the cotreatment of adriamycin and herb extract. For a while, the expression of Sax was a eminently increased by the ethanol extract of herb. However, Scl2 expression was not affected by the individual or cotreatment of adriamycin and herb extract. Conclusions : our results suggest that the cotreatment of adriamycin aM ethanol extract of herb induces synergistic apoptotis of human liver origin Chang cells via the upregulation of JNK, Fas, FasL and Bax.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.23
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• pp.10513-10524
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• 2015

Background: Metastasis is a major reason for poor prognosis in patients with cancer, including hepatocellular carcinoma (HCC). A salient feature is the ability of cancer cells to colonize different organs. Long non-coding RNAs (lncRNAs) play important roles in numerous cellular processes, including metastasis. Materials and Methods: In this study, the lncRNA expression profiles of two HCC cell lines, one with high potential for metastasis to the lung (HCCLM3) and the other to lymph nodes (HCCLYM-H2) were assessed using the Arraystar Human LncRNA Array v2.0, which contains 33,045 lncRNAs and 30,215 mRNAs. Coding-non-coding gene co-expression (CNC) networks were constructed and gene set enrichment analysis (GSEA) was performed to identify lncRNAs with potential functions in organ-specific metastasis. Levels of two representative lncRNAs and one representative mRNA, RP5-1014O16.1, lincRNA-TSPAN8 and TSPAN8, were further detected in HCC cell lines with differing metastasis potential by qRT-PCR. Results: Using microarray data, we identified 1,482 lncRNAs and 1,629 mRNAs that were differentially expressed (${\geq}1.5$ fold-change) between the two HCC cell lines. The most upregulated lncRNAs in H2 were RP11-672F9.1, RP5-1014O16.1, and RP11-501G6.1, while the most downregulated ones were lincRNA-TSPAN8, lincRNA-CALCA, C14orf132, NCRNA00173, and CR613944. The most upregulated mRNAs in H2 were C15orf48, PSG2, and PSG8, while the most downregulated ones were CALCB, CD81, CD24, TSPAN8, and SOST. Among them, lincRNA-TSPAN8 and TSPAN8 were found highly expressed in high lung metastatic potential HCC cells, while lowly expressed in no or low lung metastatic potential HCC cells. RP5-1014O16.1 was highly expressed in high lymphatic metastatic potential HCC cell lines, while lowly expressed in no lymphatic metastatic potential HCC cell lines. Conclusions: We provide the first detailed description of lncRNA expression profiles related to organ-specific metastasis in HCC. We demonstrated that a large number of lncRNAs may play important roles in driving HCC cells to metastasize to different sites; these lncRNAs may provide novel molecular biomarkers and offer a new basis for combating metastasis in HCC cases.

• The Journal of Internal Korean Medicine
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• v.23 no.1
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• pp.57-64
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• 2002

Objective : This study was designed to investigate the anti-oxidative effects of Oldenlandiae Diffusae Herba Water extract (ODHW) on lipid peroxidation by free radicals oxidative hepatic injury. Methods : In order to evaluate anti-oxidative activities of ODHW in the liver cell, cultured normal rat liver cells(Ac2F) were incubated with or without ODHW. After 16 hours to 18 hours of experiment, cells were placed in DMEM medium without serum, and then incubated with 1mM tert-butyl hydro-peroxide(t-BHP) for two hours. Viable cells were detected by MTT assay. The levels of LPO induced by hydroxyl radical derived from H2O2-Fe2+ system in rat liver homogenate were determined by means of TBA. Inhibitory effect of ODHW on superoxide generation was measured by xanthine-xanthine oxidase system. Results : In the linoleic acid autoxidation system, ODHW exhibited antioxidant activity, which inhibited 85% of linoleic acid peroxidation. These effects were similar to those of dl-a-tocopherol. ODHW showed scavenging effects on DPPH radical, inhibited superoxide generation in xanthine-xanthine oxidase system, and also inhibited lipid peroxidation of rat liver tissue with hydroxyl radical derived from $H_2O_2-Fe^{2+}$ system. In addition, ODHW protected the cell death induced by t-BHP and it significantly increased cell viability in a normal rat liver cell(Ac2F)

• The Journal of Internal Korean Medicine
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• v.29 no.2
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• pp.299-310
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• 2008

Objectives : This study was performed to investigate the anti-fibrogenic effect of Salvia miltiorrhiza on rat hepatic stellate cells. Materials and Methods : Hepatic stellate cells(HSC-T6) were treated with various concentrations of Salvia miltiorrhiza extract for 24 hours. It was extracted either with distilled water or 50% EtOH. After the treatment, cell viability, proliferation, procollagen levels and the mRNA of the collagen type 1a2 and ASMA were measured Results : The viability and proliferation of the hepatic stellate cells were decreased as concentration increased. The mRNA expression decreased consistently with the volume of the secreted procollagen with the extraction made with distilled water. This indicates the herb has inhibitory effect on fibrogenesis of the liver by regulating one of the fibrosis associated genes in transcription. However it increased in 50% EtOH extraction, which shows that a more stable reaction is expected of the extraction made with distilled water than the extraction made with 50% EtOH. The production of procollagen was decreased by a low-concentration treatment with Salvia miltiorrhiza, but increased by a high concentration. It seemed that the cells were responding to Salvia miltiorrhiza in low- concentration, thus producing smaller amounts of collagen. When the drug was administered in high enough concentration to give direct toxicity to cells, an overproduction of collagen was observed. Conclusion : These results suggest that Salvia miltiorrhiza is a possible candidate for the treatment of cirrhotic patients as well as for patients with chronic hepatitis when extracted with water in the proper concentrations.

• Journal of the Korean Society of Food Science and Nutrition
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• v.23 no.3
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• pp.380-386
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• 1994

The behavior of glycogen and histological changes of hepatic tissues in the liver of rats, aged 6 to 7 weeks, fed 18% casein diet under control, gramoxone and gramoxone + vitamin C(Vt. C) diets has been investigated in a comined histropathological and histochemical studies. Cloudy swelling and fat changes of hepatic cells were observed in the gramoxone group with the duration of feeding time. Fat changes of hepatic cells were observed more obviously than cloudy welling, especially in the hepatic cells of periportal area. The number of Kupffer's cells increased signifciantly in the gramoxone group fed for 4weeks. The cloudy swelling and fat changes decreased obviously in the gramxone + Vt. C group. Glycogen content of heaptic cells tended to increase slightly in the gramoxone group as compared with the control group . moreover, glycogen depositons were higher in the hepatic cells where fat changes were obvious. It seems to be that Vt.C alleviating effects on the gramoxone toxicity in the patterns of glycogen distribution and histological structure of heaptic tissues.

• Molecules and Cells
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• v.38 no.5
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• pp.457-465
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• 2015

The 15-kDa selenoprotein (Sep15) is a selenoprotein residing in the lumen of the endoplasmic reticulum (ER) and implicated in quality control of protein folding. Herein, we established an inducible RNAi cell line that targets Sep15 mRNA in Chang liver cells. RNAi-induced Sep15 deficiency led to inhibition of cell proliferation, whereas cell growth was resumed after removal of the knockdown inducer. Sep15-deficient cells were arrested at the G1 phase by upregulating p21 and p27, and these cells were also characterized by ER stress. In addition, Sep15 deficiency led to the relocation of focal adhesions to the periphery of the cell basement and to the decrease of the migratory and invasive ability. All these changes were reversible depending on Sep15 status. Rescuing the knockdown state by expressing a silent mutant Sep15 mRNA that is resistant to siRNA also reversed the phenotypic changes. Our results suggest that SEP15 plays important roles in the regulation of the G1 phase during the cell cycle as well as in cell motility in Chang liver cells, and that this selenoprotein offers a novel functional link between the cell cycle and cell motility.

• The Journal of the Korean Society for Microbiology
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• v.10 no.1
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• pp.39-58
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• 1975

These studies were carried out to detect the presence of mycotoxin producing fungi in various kind of grains and foodstuffs in Korea. The experiments were divided into three parts: bacteriologic, toxicologic and electron microscopic studies. The results were summarized as follows: 1. From the 133 various samples, 426 colonies of fungi were isolated. In 405 of 426 colonies, it was possible to identify 17 genera. Among the identified strains, the predominant genera were Penicillium, Aspergillus and Alternaria. 2. In cytotoxicity test, 20 strains showed mild to severe toxic effects in mice and 24 strains showed toxic effects on HeLa cells among the 107 strains of experiments. 3. In electron microscopic studies of liver cells from animal which had been treated with toxin like substances, the liver cells showed the cytoplasmic changes: dilatation of endoplasmic reticulum, swelling of mitochondria, disappearance of mitochondrial cristae, increased number of lipid and glycogen particles. Nucleus and nuclear envelope alterations were also noted. 4. In fine structure of HeLa cells treated with culture filtrates of mycotoxin producing fungi and experimental strains had been noted a certain specific changes induced by culture filtrates. These alterations showed the disappearance of golgi complex, endoplasmic reticulum vacuolization and mitochondrial changes. 5. As a mass screening, the cytotoxicity tests of HeLa cells and histopathologic study of mice liver cells treated with toxin-like substances, employed are feasible to detect mycotoxin producing fungi.