• Title/Summary/Keyword: Hep G2 cell

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Identification of the novel substrates for caspase-6 in apoptosis using proteomic approaches

  • Cho, Jin Hwa;Lee, Phil Young;Son, Woo-Chan;Chi, Seung-Wook;Park, Byoung Chul;Kim, Jeong-Hoon;Park, Sung Goo
    • BMB Reports
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    • v.46 no.12
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    • pp.588-593
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    • 2013
  • Apoptosis, programmed cell death, is a process involved in the development and maintenance of cell homeostasis in multicellular organisms. It is typically accompanied by the activation of a class of cysteine proteases called caspases. Apoptotic caspases are classified into the initiator caspases and the executioner caspases, according to the stage of their action in apoptotic processes. Although caspase-3, a typical executioner caspase, has been studied for its mechanism and substrates, little is known of caspase-6, one of the executioner caspases. To understand the biological functions of caspase-6, we performed proteomics analyses, to seek for novel caspase-6 substrates, using recombinant caspase-6 and HepG2 extract. Consequently, 34 different candidate proteins were identified, through 2-dimensional electrophoresis/MALDI-TOF analyses. Of these identified proteins, 8 proteins were validated with in vitro and in vivo cleavage assay. Herein, we report that HAUSP, Kinesin5B, GEP100, SDCCAG3 and PARD3 are novel substrates for caspase-6 during apoptosis.

Inhibitory Effects of Fungal Metabolites Isolated from Foodstuffs on the Growth of Human Cancer Cell Lines (식품유래 곰팡이 대사산물의 항암효과)

  • Im, Hyo-Gwon;Yu, Mi-Hee;Chung, Duck-Wha;Lee, In-Seon
    • Korean Journal of Food Science and Technology
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    • v.38 no.2
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    • pp.262-267
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    • 2006
  • Inhibitory effects of fungal metabolites isolated from foodstuffs on growth of human cancer cell lines were evaluated. Isolated strains were divided into four classes based on color (aerial, reverse), shape, and growth speed. Fungal metabolites extracted with ethyl acetate were investigated for their growth inhibition on six kinds of human cancer cells by MTT assay. Ethyl acetate extract showed high growth inhibition against all cancer cells tested, with D4 exhibiting the strongest growth inhibition effects against Kato III, AGS, Hepa1c1c7, and MDA-MB-231 cancer cells. These results suggest ethyl acetate extract from fungal metabolites as effective natural cancer therapeutic material.

Antimutagenic and Antitumor Effects of Adenophora triphylla Extracts (잔대 추출물들의 항돌연변이 및 항종양 효과)

  • Ham, Young-An;Choi, Hyun-Jin;Kim, Soo-Hyun;Chung, Mi-Ja;Ham, Seung-Shi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.1
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    • pp.25-31
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    • 2009
  • This study was carried out to investigate the mutagenic, antimutagenic, cytotoxicity and antitumor effects of Adenophora triphylla (AT). AT was extracted with 70% ethanol and then further fractionated to hexane, chloroform, ethyl acetate, butanol and water. Antimutagenic, cytotoxicity and antitumor effects of AT extracts were measured by using Ames test, SRB method, and the tumor growth inhibition test. AT extracts did not show any mutagenicity in the Ames test; however, 70% ethanol extracts and its fractions had strong antimutagenic effects against mutation induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and 4-nitroquinoline-1-oxide (4NQO). The ethyl acetate fraction of AT (200 ${\mu}g$/plate) showed approximately 66.5% inhibitory effect on the mutagenesis induced by 4NQO against TA98 strain, whereas 83.3% and 75.1% inhibitions were observed on the mutagenesis induced by MNNG and 4NQO against TA100 strain. In anticancer effects, the cytotoxicity of AT extract and its fractions against cancer cell lines including human cervical adenocarcinoma (HeLa), human hepatocellular carcinoma (Hep3B), human breast adenocarcinoma (MCF-7), human gastric carcinoma (AGS), human lung carcinoma (A549) and transformed primary human embryo kidney (293) were investigated. The treatment of 1 mg/mL AT ethyl acetate faction had the highest cytotoxicity of 79.9%, 74.9%, 66.0%, 71.0% and 74.3% against HeLa, Hep3B, MCF-7, AGS and A549 cells, respectively. In contrast, the extract and its fractions showed only $3{\sim}36%$ cytotoxicity for a normal human kidney cell line (293). In vivo anti-cancer effect of Adenophora triphylla extract was tested using Balb/c mice transplanted sarcoma-180 cells. Adenophora triphylla ethyl acetate fraction showed the highest inhibition rate of 37.2% at the 50 mg/kg concentration.

Immune Activities in Hypericum perforatum L. (고추나물의 면역 활성)

  • Park, Jin-Hong;Kim, Dae-Ho;Choi, Geun-Pyo;Ryu, Lee-Ha;Lee, Kang-Yoon;Lee, Hyeon-Yong
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.4
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    • pp.304-308
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    • 2004
  • Immune enhancing activities of water and ethanol extracts of Hypericum perforatum L. (HP) were examined. HP extracts inhibited the growth of human hepatocarcinoma, human gastric cancer cell and human breast cancer cells in concentration-dependent mammers over a concentration range of $0.05{\sim}1.0\;mg/ml$, showing inhibiton of more than 80% with the concentration of 1.0 mg/ml. However, HP the same concentration. Overall selectivity of the extracts on the three human cancer lines was over 3.5, which is higher than those from the conventional herbs. The growth of human immune B and T cells was enhanced up to 1.4 to 2.0 folds by the addition of the extracts for 4 days, compared to controls. Ethanol extracts of HP after 6 days incubation increased the secretions of tumor necrosis factor-alpha $(TNF-{\alpha})$ from T cells and interleukin-6 (IL-6) from B cells to 6.7 pg/cell and 6.8 pg/cell, respectively. These results suggest that HP has a potent immune enhancing effect.

Antioxidant and Antigenotoxic Effects of Shiitake Mushrooms Affected by Different Drying Methods (건조 방법에 따른 표고버섯의 항산화능과 항유전독성 효과)

  • Kim, Min-Jung;Chu, Won-Mi;Park, Eun-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.8
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    • pp.1041-1048
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    • 2012
  • Shiitake mushroom (SM; Lentinus edodes) are cultivated and consumed in many Asian countries including Vietnam, China, Japan, Korea, and Thailand. In Asia, SM are mainly dried and used as flavoring. The aim of this study was to compare the effects of SM created with different drying processes, such as oven-dried and sun-dried, on the antioxidative and antigenotoxic effects. Raw and dried SM were extracted with acetone, ethanol, methanol, and hot water. The antioxidant effects of SM were evaluated by determining total phenolic content, DPPH radical scavenging activity (RSA), an ORAC assay, and a cellular antioxidant capacity (CAC) assay. The inhibitory effect of SM on oxidative stress-induced DNA damage in human leukocytes was evaluated by a Comet assay. The total phenolic content of raw SM extracted with methanol and of that extracted with water were significantly higher than the dried SM. Among the water extracts, the $IC_{50}$ for DPPH RSA of raw and sun-dried SM were significantly higher than that of oven-dried SM. Sun-dried SM showed the most potent ORAC value at 50 g/mL. The CAC against $AAPH^-$ induced oxidative stress in HepG2 cells, and $H_2O_2$ induced DNA damage were effectively protected against by all SM extracts. These results suggest that unprocessed SM are the best antioxidants, and that the sun-dried method would be the best option to use in terms of antioxidant activity and the antigenotoxic effect.

Comparison of Anti-Oxidative Activities of Perilla frutescens Extracts by Extraction Methods (추출 방법에 따른 자소엽 추출물의 항산화 효과 비교)

  • Seo, In-Yeong;Kim, Hee-Su;Jang, Kyeong-Su;Yeo, Min-Ho;Kim, Hye-Ran;Jung, Bo-Kyoung;Chang, Kyung-Soo
    • Journal of the Korean Applied Science and Technology
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    • v.35 no.1
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    • pp.12-19
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    • 2018
  • Perilla frutescens (P. frutescens) is one of evergreen shrubs belonging to the Labiatae and is grown wildly in Korea. This study was carried out to evaluate the anti-oxidative effects of Perilla frutescens Extracts by Extraction Methods (water, heating and sonication). Anti-oxidative effects were measured using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and total phenol content. Cell viability and hepatoprotective effects were identified by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay. Among various extracts, P. rutescens extracts by sonication showed the highest DPPH radical scavenging activity at $5000{\mu}g/mL$. Total phenolic content in P. frutescens extracts by sonication was $51.60{\pm}1.06mg\;GAE/g$ extract. However, P. frutescens extracts did not show hepatoprotective effects. This study identified anti-oxidative effects of P. frutescens extracts by sonication, and it would be necessary to perform further studies of P. frutescens extracts by sonication.

Antimutagenic and Antitumor Effects of Codonopsis lanceolata Extracts (더덕 추출물의 항돌연변이 및 항종양 효과)

  • Kim, Soo-Hyun;Choi, Hyun-Jin;Chung, Mi-Ja;Cui, Cheng-Bi;Ham, Seung-Shi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.10
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    • pp.1295-1301
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    • 2009
  • This study was carried out to investigate the mutagenic, antimutagenic, cytotoxicity and antitumor effect of Codonopsis lanceolata (CL). CL was extracted with 70% ethanol and then further fractionated to hexane, chloroform, ethyl acetate, butanol and water. Antimutagenic, cytotoxicity and antitumor effects of CL extracts were measured by using Ames test, SRB method, and the tumor growth inhibition test. CL extracts did not show any mutagenicity in the Ames test; however, 70% ethanol extracts and its fractions had strong antimutagenic effects against mutation induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and 4-nitroquinoline-1-oxide (4NQO). The ethyl acetate fraction of CL (200 ${\mu}g$/plate) showed approximately 72.1% inhibitory effect on the mutagenesis induced by 4NQO against TA98 strain, whereas 69.6% and 67.0% inhibitions were observed on the mutagenesis induced by MNNG and 4NQO against TA100 strain. In anticancer effects, the cytotoxicity of CL extract and its fractions against cancer cell lines including human cervical adenocarcinoma (HeLa), human hepatocellular carcinoma (HepG2), human breast adenocarcinoma (MCF-7), human lung carcinoma (A549) and transformed primary human embryo kidney (293) were investigated. The treatment of 1 mg/mL CL ethyl acetate fraction had the highest cytotoxicity of 74.5%, 70.7% and 80.3% against HeLa, MCF-7 and A549 cells, respectively. In contrast, the extract and its fractions showed only 2$\sim$31% cytotoxicity for a normal human kidney cell line (293). In vivo anticancer effect of CL extract was tested using Balb/c mice transplanted sarcoma-180 cells. CL ethyl acetate fraction showed the highest inhibition rate of 56.4% at the 50 mg/kg concentration.

Changes in the Physiological Activities of Four Sweet Potato Varieties by Cooking Condition (고구마 네 가지 품종의 조리방법에 따른 생리활성 변화)

  • Lee, Young-Min;Bae, Ji-Hyun;Kim, Jung-Bong;Kim, So-Young;Chung, Mi-Nam;Park, Mi-Young;Ko, Jeong-Sook;Song, Jin;Kim, Jae-Hyun
    • Journal of Nutrition and Health
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    • v.45 no.1
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    • pp.12-19
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    • 2012
  • The present study was performed to investigate antioxidant, anticancer, and antimicrobial activities of four Korean sweet potato variaties and to identify the changes in these biological activities under different cooking conditions. Total polyphenol content was 3.8-73.6 mg/g in 80% ethanol extracts of sweet potatoes. The polyphenol content was highest Sinjami variety (p < 0.05). Radical scavenging activity against DPPH and $ABTS^{{\cdot}+}$ was high in Sinjami (p < 0.05) and the ethanol extract from Sinjami also showed effective superoxide dismutase (SOD)-like activity, which decreased significantly by steaming and roasting (p < 0.05). Ethanol extracts from the four sweet potato variaties did not inhibit cancer cell growth in MCF-7 or HepG2 cells at concentrations of 1, 10, and $100\;{\mu}g$/mL. Of the investigated sweet potato variaties, only Sinjami exhibited strong antimicrobial activity against Escherichia coli, Staphylococcus aureus, and Salmonella typhimurium. The antimicrobial activity of Sinjami against E. coli, St. aureus, and S. typhimurium decreased following steaming and roasting (p < 0.05). These results indicate that the Sinjami Korean sweet potato had higher polyphenol content, radical scavenging activity, SOD-like activity, and antimicrobial activity than those of the other variaties and consuming raw Sinjami might be beneficial for maintenance of biological activities.

Galectin-9 Acts as a Prognostic Factor with Antimetastatic Potential in Hepatocellular Carcinoma

  • Zhang, Zhao-Yang;Dong, Jia-Hong;Chen, Yong-Wei;Wang, Xian-Qiang;Li, Chong-Hui;Wang, Jian;Wang, Guo-Qiang;Li, Hai-Lin;Wang, Xue-Dong
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.6
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    • pp.2503-2509
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    • 2012
  • Considerable research has been conducted concerning galectin-9 and carcinomas, but little information is available about any relation with the hepatocellular carcinoma. In this study, we employed a small interfering RNA (siRNA) targeting galectin-9 to down-regulate the expression in HepG2 cells. As a result, after galectin-9 expression was reduced, cell aggregation was suppressed, while other behaviour such as the proliferation, adhesion and invasion to ECM, cell-endothelial adhesion and transendothelial invasion of the cells were markedly enhanced. When tumors of 200 patients with hepatocellular carcinoma were tested for galectin-9 expression by immunohistochemistry, binding levels demonstrated intimate correlations with the histopathologic grade, lymph node metastasis, vascular invasion and intrahepatic metastasis (P<0.05). Moreover, survival analysis indicated that patients with galectin-9 expression had much longer survival time than those with negative lesions, and the Log-rank test indicated that this difference was statistical significant (P<0.0001). The Cox proportional hazards model suggested that negative galectin-9 expression in hepatocellular carcinoma represented a significant risk factor for patient survival. We propose that galectin-9 might be a new prognostic factor with antimetastatic potential in patients with hepatocellular carcinoma.

Cytotoxicity of Methanol Extracts from Cudrania tricuspidata Bureau (꾸지뽕나무 메탄올 추출물의 세포독성)

  • Choi, So-Ra;You, Dong-Hyun;Jang, Ik;Ahn, Min-Sil;Song, Eun-Ju;Seo, Sang-Young;Choi, Min-Kyung;Kim, Young-Sun;Kim, Myung-Kon;Choi, Dong-Geun
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.3
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    • pp.153-158
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    • 2012
  • In order to develop as a natural source of anticancer materials of Cudrania tricuspidata, the cytotoxicity of methanol extracts by harvesting parts and times against 8 cell lines including 293 (normal kidney cells) and A-431 (epidermoid carcinoma cells) were investigated using MTT assay. All harvesting parts had hardly cytotoxicity against 293. And methanol extracts of stem bark and root bark showed very high cytotoxicities against 7 cancer cell lines. The cytotoxicity was the highest against HeLa (cervix adenocarcinoma cells) and followed by MCF-7 (breast adenocarcinoma cells), AGS (stomach adenocarcinoma cells), HT-29 (colon adenocarcinoma cells), HepG2 (hepatoblastoma cells), A549 (lung carcinoma cells) and A-431. By the way, leaf extract had a cytotoxicity against only AGS and ripe fruit extract had no cytotoxicity. Among harvesting times, the cytotoxicity of root bark were high from April to September but that of stem bark showed a little difference. These results showed that anticancer activities of Cudrania tricuspidata extracts were eventful changes by harvesting parts and times.