• Title/Summary/Keyword: HEP 2

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Inhibitory Mechanisms of Cell Cycle Regulation Induced by Indole-3-carbinol in Hepatocellular Carci-noma HepG2 Cells. (간암 세포주에서의 Indole-3-Carbinol에 의해 유도되는 세포주기 억제 기전)

  • 김동우;이광수;김민경;조율희;이철훈
    • Microbiology and Biotechnology Letters
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    • v.29 no.3
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    • pp.181-185
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    • 2001
  • The naturally occurring chemical indole-3-carbinol (13C), found in vegetables of the Brassica genus, is a promising anticancer agent that was shown previ- ously to induce a Gl cell cycle arrest of human breast cancer cell lines, independent of estrogen receptor signaling. The anticancer activity of 13C and the possible mechanisms of its action were explored in a human hepatocellular carcinoma cell line, HepG2. Treatment of HepG2 cells with 13C suppressed the growth of the cells. The growth sup- pression caused by 13C ($IC_{50}$/: 444$\mu$M) was found to be partially due to its ability to stop the cell cycle in HepG2 cells. Western blot analysis for the Gl phase artiest demonstrated that the expression-levels of cyclin-dependent kinase (Cdk4, Cdk6) and cyclic D were reduced strongly after treatment of Hep72 cells with 13C (4007M) for 24- 72 hrs. Furthermore, I3C selectively abolished the expression of Cdk6 in a dose- and time-dependent manner, and accordingly, inhibited the phosphorylation of retinoblastoma. Interestingly, after the HepG2 cells reached their max- imal growth arrest, the level of the p21, a well-known Cdk inhibitor, increased significantly. Therefore, it could be considered that the Gl arrest of HepG2 cells treated with 13C was due to the indirect inhibition of Cdk4/6 activities by p21 Western blot analysis for G2/M phase arrest of demonstrated the levels of Cdc2 and cyclin Bl werer reduced dramatically after the treatment of HepG2 cells with 13C ($40\mu$M) for 24-72 hrs. flow cytometry of propidium iodide-stained HepG2 cells revealed that 13C induces a Gl (53%,72hr incubation) and G2 (25%,24hr incubation) cell cycle arrest. Thus, our observations have uncovered a previously undefined antiproliferative pathway for r3C that implicates Cdk4/6 and Cdc2 as a target for cell cycle control in human HepG2 cells. However, the 13C-medi- ated cell cycle arrest and repression of Cdk4/6 production did not affect the apoptotic induction of HepG2 cell.

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Antioxidative and Anticancer Activities of Ethanol Extract of Millettia erythrocalyx (Millettia erythrocalyx 에탄올 추출물의 항산화 활성 및 항암 활성에 관한 연구)

  • Jin, Soojung;Oh, You Na;Son, Yu Ri;Choi, Sun Mi;Kwon, Hyun Ju;Kim, Byung Woo
    • Journal of Life Science
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    • v.28 no.1
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    • pp.50-57
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    • 2018
  • Millettia erythrocalyx, a species of plant in the Fabaceae family, is widely distributed in the tropical and subtropical regions of the world, such as the Indies, China, and Thailand. The antiviral activity of flavonoids from M. erythrocalyx has been reported; however, the antioxidative and anticancer activities of M. erythrocalyx remain unclear. In this study, we evaluated the antioxidative and anticancer effects of ethanol extract of M. erythrocalyx (EEME) and the molecular mechanism of its anticancer activity in human hepatocellular carcinoma HepG2 cells. EEME exhibited significant antioxidative effects, with a concentration at 50% inhibition ($IC_{50}$) value of $2.74{\mu}g/ml$, as measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay; moreover, it inhibited cell proliferation in a dose-dependent manner in HepG2 cells. Cell cycle analyses showed that EEME induced HepG2 cell accumulation in the subG1 phase in a dose-dependent manner. EEME also induced apoptosis of HepG2 cells, with increases in apoptotic cells and apoptotic bodies, as detected by Annexin V and 4,6-diamidino-2-phenylindole (DAPI) staining, respectively. Treatment with EEME resulted in increased expression of First apoptosis signal (Fas), a death receptor, and Bcl-2-associated X protein (Bax), a proapoptotic protein, and the activation of caspase-3, 8, and 9, resulting in the cleavage of poly (Adenosine diphosphate-ribose) polymerase (PARP). Collectively, these results suggest that EEME may exert an anticancer effect in HepG2 cells by inducing apoptosis via both the intrinsic and extrinsic pathways.

Cooperative stimulation of cisplatin-mediated apoptosis by hepatitis B virus X Protein and hepatitis C virus core Protein (B형 간염 바이러스 X 단백질과 C형 간염 바이러스의 코어 단백질에 의한 cisplatin-매개성 세포 예정사의 협조적 촉진)

  • Kwun, Hyun-Jin;Jang, Kyung-Lib
    • Journal of Life Science
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    • v.17 no.6 s.86
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    • pp.766-771
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    • 2007
  • The co-infection with hepatitis B virus (HBV) and hepatitis C Virus (HCV) is associated with a more severe liver disease and increased frequency in the development of hepatocellular carcinoma com-pared to those with single infection. Here, we demonstrated that HBV X protein (HBx) and HCV Core cooperatively up-regulated the level of p53 in human hepatoma HepG2 cells. The elevated p53 subsequently stimulated the expression of proapoptotic Bax whereas it repressed the expression of antiapoptotic Bcl2. These effects, however, were not observed in p53-negative Hep3B cells. Consistently to their cooperative regulation of apoptotic effectors, HBx and HCV Core additively stimulated cisplatin-mediated apoptotic cell death of HepG2 but not of Hep3B cells. These results may help to explain the development of a more severe liver disease in patients co-infection with HBV and HCV as well as some contradictory results on the roles of HBx and Core in apoptosis.

Selective Cytotoxic Effects of Doenjang (Korean Soybean Paste) Fermented with Bacillus Strains on Human Liver Cell Lines

  • Choi, Myeong-Rak;Lim, Hyun-Soo;Chung, Yoon-Ju;Yoo, Eun-Jeong;Kim, Jong-Kyu
    • Journal of Microbiology and Biotechnology
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    • v.9 no.4
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    • pp.504-508
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    • 1999
  • This report compares the selective cytotoxic effects of Doenjang fermented by various Bacillus strains (Bacillus sp. SS9, SSA3, and PM3) on human liver cell lines with that of conventional Doenjang (DTY, DTG, and DTK) and commercial Doenjang (DCM, DCD, and DCS). To investigate selective cytotoxic effects of Doenjang extracts, the cell density of HepG2 (Hepatocellular carcinoma) and CCL-13 (cells derived from human normal liver) was estimated after addition of the extracts by using a viable cell counting method. The maximum selectivity ratio ($IC_{50}$value against CCL-13/$IC_{50}$ value aganist HepG2) was observed by PM3 (extracts of Doenjang fermented with Bacillus sp. PM3). As for morphological changes shown by the addition of PM3 into HepG2 and CCL-13 cultures, HepG2 was significantly disrupted, however, CCL-13 was not affected. Also, the growth rate of HepG2 was decreased significantly by the addition of PM3. Consequently, PM3 showed a more detrimental effect on HepG2 than that on CCL-13.

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The Effects of Injinchunggan-tang(Yinchenqinggan-tang) on Expression of Angiogenic Factors in HepG2 Cells (인진청간탕(茵蔯淸肝湯)이 인체 간암세포의 혈관생성인자 발현에 미치는 영향)

  • Kim, Chul-Woo;Kim, Young-Chul;Lee, Jang-Hoon;Woo, Hong-Jung
    • The Journal of Internal Korean Medicine
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    • v.27 no.1
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    • pp.138-148
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    • 2006
  • Objectives: This study was designed to investigate the effects of Injinchunggan-tang(Yinchenqinggan-tang) on expression of angiogenic factors in HepG2 cells. Materials and Methods : The mRNA expression levels and protein secretion levels of angiogenic factors were measured using quantitative RT-PCR, Western blot and ELISA assay respectively in Injinchunggan-tang-treated and untreated HepG2 cells. Results : Injinchunggan-tang(Yinchenqinggan-tang) reduced mRNA expression levels and protein secretion levels of angiogenic factors, especially VEGF, bFGF and $TGF{\beta}1$ in HepG2 cells. Conclusion: Results indicate that Injinchunggan-tang (Yinchenqinggan-tang) inhibits expression of angiogenic factors in HepG2 cells. Further, results suggest that Injinchunggan-tang (Yinchenqinggan-tang) inhibits angiogenic effects in HCC.

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A Probing of Inhibition Effect on Specific Interaction Between Glucose Ligand Carrying Polymer and HepG2 Cells

  • Park, Keun-Hong;Park, Sang-Hyug;Lee, Hyun-Jung;Min, Byoung-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.450-455
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    • 2004
  • A reducing glucose-carrying polymer, called poly [3-O-(4'-vinylbenzyl)-D-glucose](PVG), was interacted with HepG2 cells including a type-l glucose transporter (GLUT-1) on the cell membrane. The cooperative interaction between a number of GLUT-1s and a number of reducing 3-O-methyl-D-glucose moieties on the PVG polymer chain was found to be responsible for the increase in the interaction with HepG2 cells. The affinity between the cells and the PVG was studied using RITC-labeled glycopolymers. The specific interaction between the GLUT-1 on HepG2 cells and the PVG polymer carrying reducing glucose moieties was suppressed by the inhibitors, phloretin, phloridzin, and cytochalasin B. Direct observation by confocal laser microscopy with the use of RITC-labeled PVG and pretreatment of HepG2 cells with the inhibitors demonstrated that the cells interacted with the soluble form of the PVG polymer via GLUT-1, while fluorescence labeling of the cell surface was prevented after pretreatment with the inhibitors of GLUT-1.

The Effects of Loranthus parasiticus Merr. on Cell Cycle and Expression of Related Genes in HepG2 Cell (상기생(桑寄生)이 HepG2 cell의 세포분열 및 관련유전자 발현에 미치는 영향)

  • Rhew, Kwang-Yul;Kim, Young-Chul;Woo, Hong-Jung;Lee, Jang-Hoon
    • The Journal of Internal Korean Medicine
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    • v.26 no.1
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    • pp.60-73
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    • 2005
  • Objectives : The aim of this study was to evaluate the effects of Loranthus parasiticus Merr. on cell cycle and expression of related genes in HepG2 cells. Methods : The MTT assay, cell counting assay, $[^3H]-Thymidine$ incorporation assay, flow cytometric analysis, quantitative RT-PCR and western blot assay were studied. Results : In the water extract of Loranthus parasiticus Merr., inhibition of cell proliferation and DNA synthesis in HepG2 cells was seen. These inhibitory effects were due to inhibition of G l-S transition in cell cycle. After treatment with the extract, expression of cyclin D1(G1 check point related gene) was inhibited particularly in dose-dependent and time-dependent manners. Conclusion : These results suggest that the inhibition of cell cycle progression by Loranthus parasiticus Merr. in HepG2 cell is due to suppression of cyclin D1(G1 check point related gene) mRNA expression and protein synthesis.

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Effect of CLA (Conjugated Linoleic Acid) on the Anti-Atherosclerotic actors in Human Hepatoma HepG2 Cells (간암세포 (HepG2 Cell)에서의 식이성 CLA(Conjugated Linoleic Acid)가 항동맥경화성 인자에 미치는 영향)

  • 오현희;문희정;이명숙
    • Journal of Nutrition and Health
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    • v.37 no.3
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    • pp.182-192
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    • 2004
  • Conjugated linoleic acid (CLA) is the mixture of positional and geometric isomers of linoleic acid (LA), which is found abundantly in dairy products and meats. This study was performed to investigate the anticarcinogenic effect of CLA in HepG2 hepatoma cells. HepG2 cell were treated with LA and CLA at the various concentrations of 10, 20, 40, 80 uM each at different incubation times. After each incubation times, cell proliferation, fatty acids incorporation into cell, peroxidation and postaglandin E$_2$ (PGE$_2$) and thromboxane $A_2$ (TXA$_2$) for the eicosanoid metabolism were measured. LA treated HepG2 cells were increased cell growth 6 - 70% of control whereas CLA increased cell death the half of those in LA group (p 〈 0.001). LA and CLA were incorporated very well into the cellular membranes four times higher than in control according to concentration and longer incubation times. Moreover, LA synthesized significantly arachidonic acids corresponding with LA concentration compared to CLA supplementation. The supplementation with LA increased intracellular lipid peroxides concentration corresponding with LA concentration and five times higher than those in CLA significantly at any incubation times (p 〈 0.001). PGE$_2$ and TXA$_2$ levels were three to twenty times lower in condition of CLA treatments than LA, respectively. Overall, the dietary CLA might change the HepG2 cell growth by the changes of cell composition, production of lipid peroxide. Since CLA have not changed the levels of arachidonic acid of cell membrane, which was sources of eicosanoids, eicosanoid synthesis was not increased in CLA compared to LA. Our results was suggest CLA has a possibility to protect the progress of atherosclerosis because CLA does not produce lipid production and endothelial contraction factors in liver.

Antiproliferative and Antioxidative Activities of Methanol Extracts of Echinacea angustifolia (Echinacea angustifolia 메탄올 추출물의 암세포 증식억제 및 항산화 효과)

  • Lee Joon-Kyoung;Koo Seung-Ja
    • Korean journal of food and cookery science
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    • v.21 no.3 s.87
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    • pp.311-318
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    • 2005
  • Echinacea, also blown as the purple coneflower, is a herbal medicine that has been used for centuries, customarily as a treatment for the common cold, coughs, bronchitis, upper respiratory infections, and some inflammatory conditions. We investigated the effects of methanol extracts of Echinacea angustifolia on the cytotoxicity against cancer cells $(HepG_2,\;3LL,\;HL60,\;L1210)$ and antioxidative activity. From the test results, each part of Echinaceashowed a cytotoxic effect against the cancer cell lines, and this cytotoxic effect increased with increasing sample concentration. At 1.0 mg/mL concentration the relative cytotoxic activities of the flower bud, leaf, stern and root parts were $90.5\%,\;52.7\%,\;37.1\%\;and\;19.2\%$, respectively, in $HepG_2$ cells, and $75.5\%,\;93.3\%,\;81.2\%,\;and\;75.1\%$ respectively, in HL60 cells, as evaluated by MTT assay. $IC_{50}(50\%\;inhibitory\;concentration)$ of the methanol extracts of the Echinacea flower bud was 0.214 mg/mL on /$HepG_2$ cells, and that of the Echinacea leaf and root was 0.166 mg/mL and 0.210 mg/mL, respectively, on HL60 cells. After /$HepG_2$ cells were incubated for 6 days at $37^{\circ}C$ with various concentrations of each part, the cell number increased while the inhibition rate on the /$HepG_2$ cell growth decreased. The antioxidative activities of the flower bud, leaf, stem and root parts were $59.0\%$ (0.75 mg/mL), $80.76\%$ (0.5 mg/mL), $95.5\%$ (0.25mg/mL) and $98.15\%$ (0.25 mg/mL), respectively, as evaluated by electron donating ability. These results indicated that Echinacea angustifolia has strong anticancer and antioxidative effects in vitro.

The Effects of Chungganhaeju-tang(Qingganjiejiu-tang) on Alcohol induced Cytotoxicity in CYP2E1-transfected HepG2 cells (청간해주탕(淸肝解酒湯)이 CYP2E1-transfected HepG2 cell에서 알코올유발 세포독성에 미치는 영향)

  • Lee, Ji-Eun;Kim, Young-Chul;Woo, Hong-Jung;Lee, Jang-Hoon
    • The Journal of Internal Korean Medicine
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    • v.27 no.1
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    • pp.27-39
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    • 2006
  • Objectives : Induction of CYP2E1 by ethanol is believed to be one of the major mechanism by which ethanol generate a state of oxidative stress. Previous studies showed that treatment with Chungganhaeju-tang prevents hepatic inflammation and apoptosis in alcoholic liver disease. The purpose of our study is to determine if Chungganhaeju-tang can also protect against alcohol-induced cytotoxicity in CYP2E1-transfected HepG2 cells. Materials and Methods : CYP2E1-transfected HepG2 cells and control vector-transfected HepG2 cells were exposed for isx hours to Chungganhaeju-tang, and then 50 mM of ethanol was added and left for two days. Results : Ethanol significantly decreased cell viability in CYP2E1-transfected HepG2 cells and increased apoptosis. These alterations were attenuated by Chungganhaeju-tang. This was accompanied by an improvement of NF-${\kappa}B$ and Akt activation. Conclusion : These results suggest that Chungganhaeju-tang exerts inhibitory effect against the cytotoxicity induced by alcohol in CYP2E1-transfected HepG2 cells, and that this is a protective action due, at least in part, to an activation of NF-${\kappa}B$ that plays a key role in the protection mechanism, and in reducing hepatotoxic cytokine gene expression.

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