• Journal of Yeungnam Medical Science
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• 제37권2호
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• pp.73-78
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• 2020

Cancer incidence has been increasing steadily and is the leading cause of mortality worldwide. Gastric cancer is still most common malignancy in Korea. Cancer initiation and progression are multistep processes involving various growth factors and their ligands. Among these growth factors, we have studied hepatocyte growth factor (HGF), which is associated with cell proliferation and invasion, leading to cancer and metastasis, especially in gastric cancer. We explored the intercellular communication between HGF and other surface membrane receptors in gastric cancer cell lines. Using complimentary deoxyribonucleic acid microarray technology, we found new genes associated with HGF in the stomach cancer cell lines, NUGC-3 and MKN-28, and identified their function within the HGF pathway. The HGF/N-methyl-N'-nitroso-guanidine human osteosarcoma transforming gene (c-MET) axis interacts with several molecules including E-cadherin, urokinase plasminogen activator, KiSS-1, Jun B, and lipocalin-2. This pathway may affect cell invasion and metastasis or cell apoptosis and is therefore associated with tumorigenesis and metastasis in gastric cancer.

• 대한의생명과학회지
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• 제22권1호
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• pp.1-8
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• 2016

Several studies have investigated the various effects of dexamethasone (Dex) on the proliferation and differentiation of mesenchymal stem cells (MSCs). Previously, we reported that co-treatment with L-ascorbic acid 2-phosphate and fibroblast growth factor (FGF)-2 maintained differentiation potential in MSCs through expression of hepatocyte growth factor (HGF). In this study, we investigated the effects of co-treatment with FGF-2 and Dex on the proliferation and differentiation potential of MSCs during a 2-month culture period. Co-treatment with FGF-2 and Dex increased approximately a 4.7-fold higher accumulation rate of MSC numbers than that by FGF-2 single treatment during a 2-month culture period. Interestingly, co-treatment with FGF-2 and Dex increased expression of HGF and maintained adipogenic differentiation potential during this culture period. These results suggest that co-treatment with FGF-2 and Dex preserves the proliferation and differentiation potential during long-term culture.

• Applied Microscopy
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• 제18권2호
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• pp.205-217
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• 1988

An ultrastructural study of hepatocyte proliferation in the regenerating rat liver has been made by means of the partial hepatectomy. And electron microscopic histochemistry of hepatocyte in the regenerating rat liver is studied through alkaline phosphatase reaction. The results are as follows: 1. When the regeneration of rat liver is induced by the partial hepatectomy, the prominent ultrastructural characteristics of hepatocyte are changes of the distribution of chromatin in nucleus, increase of the number of mitochondria and decrease of the size of them, development of rough endoplasmic reticulum, and transient decrease of glycogen granules in cytoplasm. 2. Alkaline phosphatase reaction products are appeared in the nucleus or rough endoplasmic reticulum of hepatocyte during the initial regeneration of liver as 24, 48 and 72 hour groups after partial hepatectomy. And these positive reaction are mainly increased in cytoplasm and plasma membrane of hepatocytes during 1, 2 and 3 week groups after partial hepatectomy. As 4 weeks passed after partial hepatectomy, these positive reaction is located in the sinusoidal epithelial cells or erythrocytes. With above results, we concluded that alkaline phosphatase was synthesized in the rough endoplasmic reticulum bounded ribosomes of regenerating hepatocyte, was transported to the plasma membrane of them, and then was transported in blood by the way sinusoidel epithelial cells.

• Biomolecules & Therapeutics
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• 제2권3호
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• pp.236-243
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• 1994

A splenocyte culture system supplemented with liver microsomes was developed to detect immunotoxic chemicals which require metabolic activation using cyclophosphamide as a positive standard. When liver microsomes were added to splenocyte cultures isolated from female B6C3Fl mice, the proliferation of splenocytes by lipopolysaccharide (LPS) was increased and the proliferation by concanavalin A (Con A) was decreased. However, when compared with each corresponding control, cyclophophamide was successfully activated to metabolites capable of suppressing Iymphoproliferative responses. This suppression was clearly dependent upon the amounts of microsomes added and/or the concentration of cyclophosphamide exposed. In these cultures, the proliferation of splenocytes was suppressed when the cells were exposed to cyclophosphamide on the day of culture initiation. On the other hand, microsome was responsible for the increase in LPS mitogenicity and NADPH was responsible for the decrease in Con A mitogenicity. Finally, our present culture system was compared with the hepatocyte-splenocyte coculture system which we had developed earlier. We found that the hepatocyte-splenocyte coculture was better able to activate cyclophosphamide to metabolites capable of suppressing the antibody response to sheep erythrocytes. Although our present culture system was relatively poor to activate cyclophosphamide in cultures for antibody response, it will be useful as a simple screening method to detect suppression of certain in vitro immunotoxic parameters like LPS mitogenicity by chemicals which require metabolism.

• 한국동물위생학회지
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• 제31권4호
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• pp.457-463
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• 2008

The sex steroid hormone estradiol-$17{\beta}(E_2)$ mediate their biological effects on development, differentiation and maintenance of reproductive tract and other target tissue through gene regulation by nuclear steroid receptors. Although the importance of $E_2$ in many physiological process has been reported, but little is known about the effects of $E_2$ on primary cultured chicken hepatocyte. therefore, in the present study, we have examined the effect of $E_2$ on cell proliferation and it's related signal cascades. $E_2$ increase $[^3H]$-thymidine incorporation in time-(${\leq}8hr$) and dose-($10^{-10}M$)dependent manner and treatment of $E_2$ increased the phosphorylation of p44/43 MAPKs(p44/42 mitogen-activated protein kinase) and JNK(c-Jun N-terminal kinase) in a time dependent manner. In addition, PD98059(p44/42 blocker, $10^{-5}M$), SP600125(JNK blocker, $10^{-6}M$) blocked the estrogen-induced increase in $[^3H]$-thymidine incorporation. In conclusion, $E_2$ stimulates the proliferation of primary cultured chicken hepatocytes and this action is mediated by p44/42 MAPKs and JNK signal transduction pathway.

• 생명과학회지
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• 제26권10호
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• pp.1189-1195
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• 2016

Vascular endothelial growth factor (VEGF)와 hepatocyte growth factor (HGF)는 강력한 혈관신생 유도인자로 알려져 있다. 세포가 이동하고 침윤하기 위해서는 세포의 증식과 더불어 주변의 세포외기질을 분해하는 단백질분해효소의 분비가 선행되어야 한다. 본 연구에서는 인간의 악성신경교종 유래 세포주인 U-373-MG 세포에 VEGF와 HGF를 처리하여 세포의 증식, matrix metalloproteinase-2 (MMP-2)와 MMP-9 및 플라스민의 분비, 세포의 이동 및 침윤에 미치는 효과를 조사하였다. 또한 단백질분해효소 억제제 처리를 통하여 세포의 증식, 이동 및 침윤에 미치는 단백질분해효소의 역할을 조사하였다. 연구 결과, VEGF와 HGF의 병용처리 시 VEGF와 HGF의 단독 처리 시보다 세포의 증식, MMP-2, MMP-9 및 플라스민의 분비, 세포의 이동 및 침윤이 유의할만하게 증가되었다. 한편 VEGF와 HGF 처리에 의한 U-373-MG 세포의 증식, 이동 및 침윤 증가에 미치는 단백질분해효소의 효과를 MMPs 억제제인 BB-94를 처리하여 조사한 결과 최대 이동 효과를 나타낸 HGF와 VEGF의 병용처리군 보다 세포의 이동이 32% 억제되었고 플라스민 억제제인 α2AP에 의해서도 29% 억제되었다. 또한 U-373-MG 세포의 침윤 역시 BB-94와 α2AP 처리에 의해 유의할 만하게 억제되었다. 이러한 결과는 VEGF와 HGF에 의한 MMP-2, MMP-9 및 플라스민의 분비증가에 의해 직접 또는 간접적인 경로를 통하여 U-373-MG 세포의 증식, 이동 및 침윤을 증가시킨다고 여겨진다.

• Toxicological Research
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• 제8권2호
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• pp.191-203
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• 1992

The immunosuppressive effects of safrole were studied in female BALB/c mouse. Mice were given 100,200and 400mg safrole/kg daily for 14days and evaluated on day 15. The day 4 immunogloblin-M antibody response to T-dependent antigen, sheep red blood cells (SRBC) was inhibited dose-dependently in all doses studied. In vitro antibody response to polyclonal antigen, lipopolysaccharide (LPS) by spleen cell suspensions from safrole-treated mice were also significantly inhibited. When safrole was treated for 14days to mice, and mitogen-induced proliferation of splenocytes were assayed on day 15, there were significant suppression of responses to B-cell mitogen, LPS and T-cell mitogen concanavalin A(Con A) at a dose of 400mg safrole/kg. Direct addition of safrole on the splenocyte culture also produced a dose dependent suppression on in vitro antibody response to LPS, and mitogen-induced lymphoproliferatin at doses of 100,200,400 and 800${\mu}M$ safrole. The role of metabolic activation in safrole-induced suppression of in vitro antibody response was studied using splenocyte-hepatocyte coculture system. The suppression of in vitro antibody respose to LPS by safrole was not altered when safrole were incubated in the splenocyte-hepatocyte system for 4hr as compared with direct addition of safrole in splenocytes culture. Neither the addition of salicylamide, sulfotransferase inhibitor, nor the addation of inorganic sulfate, sulfation cofactor to the splenocyte-hepatocyte coculture, altered the suppression of antibody response by safrole. These results suggest that the immunosuppression by safrole may not by produced by the reactive metabolites which are mediated in carcinogenesis of safrole.

• BMB Reports
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• 제55권12호
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• pp.633-638
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• 2022

Liver regeneration is a well-known systemic homeostatic phenomenon. The N⁶-methyladenosine (m⁶A) modification pathway has been associated with liver regeneration and hepatocellular carcinoma. m⁶A methyltransferases, such as methyltransferase 3 (METTL3) and methyltransferase 14 (METTL14), are involved in the hepatocyte-specific-regenerative pathway. To illustrate the role of METTL14, secreted from non-parenchymal liver cells, in the initiation phase of liver regeneration, we performed 70% partial hepatectomy (PH) in Mettl14 heterozygous (HET) and wild-type (WT) mice. Next, we analyzed the ratio of liver weight to body weight and the expression of mitogenic stimulators derived from non-parenchymal liver cells. Furthermore, we evaluated the expression of cell cycle-related genes and the hepatocyte proliferation rate via MKI67-immunostaining. During regeneration after PH, the weight ratio was lower in Mettl14 HET mice compared to WT mice. The expressions of hepatocyte growth factor (HGF) and tumor necrosis factor (TNF)-α, mitogens derived from non-parenchymal liver cells that stimulate the cell cycle, as well as the expressions of cyclin B1 and D1, which regulate the cell cycle, and the number of MKI67-positive cells, which indicate proliferative hepatocyte in the late G1-M phase, were significantly reduced in Mettl14 HET mice 72 h after PH. Our findings demonstrate that global Mettl14 mutation may interrupt the homeostasis of liver regeneration after an acute injury like PH by restraining certain mitogens, such as HGF and TNF-α, derived from sinusoidal endothelial cells, stellate cells, and Kupffer cells. These results provide new insights into the role of METTL14 in the clinical treatment strategies of liver disease.

• 대한한의학회지
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• 제19권1호
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• pp.145-164
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• 1998

The purpose of this study is to evaluate the effect of Injinchunggantang-derivative on proliferation of hepatocyte in rats. Cell viability is studied by MTI assay. The gene related to cell replication such as p53, waf1, bcl-2 and $bcl-_{X_L}$ is quantitized by quantitative RT-PCR and the proteins coded by these genes are studied by Western blotting. The results are as follows. 1. The hepatocytes cultured in medium with lnjinchunggantang-derivative showed better viability compared with control grroup in MTI assay, and the hepatocytes cultured in medium with the Injinchunggantang-derivative-and-ethanol-mixed group showed better viability than the hepatocytes cultrued in 10% ethanol culture medium(control group), noting that Injinchunggantang-derivative has protective effect on hepatocyte injury. There was no dose- and time-dependence. 2. In quantitative RT-PCR, i) Bel-2 gene increased significantly both in Injinchunggantang-derivative group and in Injinchunggantang-derivative-and-ethanol-mixed group, while it showed no significant increase or decrease in other group. ii) $Bcl-_{X_L}$ gene increased significantly in Injinchunggantang-derivative group as well as in Injinchunggantang-deri vative-and-ethanol -mixed group. iii) P53 gene showed no significant increase or decrease in hepatocytes cultured in medium with 10% ethanol and in hepatocytes cultured in medium with Injinchunggantang-derivative-and-ethanol-mixed group, suggesting that 10% ethanol induced cell toxicity, thus increased p53 gene expression. iv) Wafl gene showed no significant increase or decrease in hepatocytes cutured in medium with Injinchtrnggantang-derivative, while increased in hepatocytes cultured in medium with 10% ethanol and in hepatocytes cultured in medium with Injinchtrnggantang-derivative-andethanol-mixed group, suggesting that 10% ethanol induced cell toxicity increased wafl gene expression. 3. In the study on protein by western blotting, the band of bcl-2 and $bcl-_{X_L}$ were widened in Injinchtrnggantang-derivative group. Especially the amount of $bcl-_{X_L}$ increased significantly compared with other groups. But in the study on p53 and wafl, there was no significant difference among those groups. Above study shows that Injinchunggantang-derivative has good effect on cell viability and that the genes resistant to cell death such as bcl-2 and $bcl-_{X_L}$ are induced by Injinchunggantang-derivative to resist to cell death by toxic agent And this is reconfirmed in protein study using' western blotting: These results suggest that Injinchunggantang-derivative has inhibitory effect on cell death as well as protective effect on hepatocyte. Therefore this prescription is recommended in various liver diseases such as chronic liver disease and-induced hepatic injury.

• KSBB Journal
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• 제24권5호
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• pp.425-431
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• 2009

본 연구에서는 U-251-MG 세포를 이용하여 증식, 이동, 침윤 및 단백질분해효소의 분비에 미치는 PSA와 HGF의 효과를 확인하였다. 그 결과, PSA siRNA에 의해 U-251-MG 세포의 증식은 약 37% 억제되었으나, HGF 처리 (10 ng/mL)에 의해 증식이 약 1.4배 증가되었다. PSA siRNA에 의한 U-251-MG 세포의 이동은 약 60%가 억제되었으나, HGF 처리에 의해 이동이 약 1.3배 증가되었다. 또한, PSA siRNA에 의해 U-251-MG 세포의 침윤이 약 67% 억제되었으나 HGF 처리에 의해 세포의 침윤이 약 4.3배 증가되었다. PSA siRNA처리에 의해 MMP-2의 분비는 약 25%, MMP-9의 분비는 약 20% 억제되었으며, HGF에 의해 PSA siRNA에 의해 억제된 MMP-2 및 MMP-9의 분비가 약 2.8배 및 3.5배 증가되었다. HGF 처리에 의해 플라스민의 분비량은 약 14배 증가되었고, PSA를 억제한 U-251-MG 세포에 HGF를 처리한 결과, 플라스민의 분비가 약 1.6배 증가하였다. 또한, PSA siRNA에 의해 MMP-2의 발현은 유의할만한 변화가 없었으나, MMP-9의 발현은 약 85% 억제되었다. PSA를 억제시킨 U-251-MG 세포에 HGF를 처리한 결과, MMP-2의 발현은 약 5.7배, MMP-9의 발현은 약 6.3배 증가되었다. 한편, MMPs의 광범위 억제제인 BB-94 처리에 의해 U-251-MG 세포의 증식, 이동 및 침윤이 유의할만하게 억제 된 것은 MMP-2 및 MMP-9이 U-251-MG 세포의 증식, 이동 및 침윤에 관여할 것임을 시사해준다.