• Title/Summary/Keyword: cancer cell lines

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The Anticarcinogenic and Antioxidative Activity of Hemicentrotus pulacherrimus Fractions in Various Cancer Cells. (말똥성게 분획물에 의한 항 발암 및 항산화 효과)

  • Shin, Mi-Ok;Bae, Song-Ja
    • Journal of Life Science
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    • v.19 no.5
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    • pp.607-614
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    • 2009
  • This study was carried out to investigate the anticarcinogenic and antioxidative activities of Hemicentrotus pulacherrimus (HP). HP was extracted with methanol (HPM), which was then further fractionated into four sub-fractions by using the solvent partition method, affording methanol (HPMM), hexane (HPMH), butanol (HPMB) and aqueous (HPMA) soluble fractions. We determined the anticarcinogenic activities of these four fractions in four kinds of cancer cell lines, such as HepG2, HT29, MCF-7 and B16-F10, by MTT assay. Among various fractions from HPM, the HPMH showed the strongest growth inhibition effect. We also determined the inductive effect on quinone reductase (QR) of HP fractions. HPMB fraction exhibited strong inductive effects in HepG2 cells at a level of 90 ${\mu}g/ml$, showing inductive indexes of 2.26 compared to the control value of 1.0. The antioxidant activities of fractions from HP were also investigated by measuring the scavenging activities of HP against reactive oxygen speicies (ROS), peroxynitrite (ONOO-) and NO. Among the various solvent fractions, HPMH fractions displayed marked antioxidative activities.

Inhibitory Potential of Thelephoric Acid on CYP2J2 Activities in Human Liver Microsomes (Thelephoric acid의 CYP2J2 효소 활성 저해제 평가)

  • Wu, Zhexue;Lee, Boram;Song, Kyung-Sik;Liu, Kwang-Hyeon
    • Journal of Life Science
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    • v.23 no.9
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    • pp.1126-1132
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    • 2013
  • Cytochrome P450 2J2 (CYP2J2) is an enzyme mainly found in human extrahepatic tissues, with predominant expression in the cardiovascular system. CYP2J2 plays important roles in the metabolism of endogenous metabolites and therapeutic drugs, such as arachidonic acid, astemizole, ebastine, and terfenadine. CYP2J2 is also overexpressed in human cancer tissues and cancer cell lines and may represent a potential target for therapy of human cancers. In this study, 10 natural products obtained from plants and microorganisms were screened as potential CYP2J2 inhibitors. Among them, thelephoric acid showed strong inhibition of astemizole O-demethylation activity ($IC_{50}=3.23{\mu}M$) in a dose-dependent manner. Evaluation of the substrate dependency of the inhibitory activity of thelephoric acid showed that it strongly inhibited CYP2J2-mediated ebastine hydroxylation ($IC_{50}=5.32{\mu}M$) and terfenadine hydroxylation ($IC_{50}=3.27{\mu}M$) in a substrate nondependent manner. The present data suggest that this compound might be a potential candidate for further evaluation for anticancer activity.

Preparation and Characterization of Anti-GP73 Monoclonal Antibodies and Development of Double-antibody Sandwich ELISA

  • Li, Qi-Wen;Chen, Hong-Bing;Li, Zhi-Yang;Shen, Peng;Qu, Li-Li;Gong, Lai-Ling;Xu, Hong-Pan;Pang, Lu;Si, Jin
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.5
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    • pp.2043-2049
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    • 2015
  • Background: Serum Golgi protein 73 (GP73) as a novel and potential marker for diagnosing hepatocellular carcinoma (HCC) have been found to be elevated in HCC patients and associated with clinical variables representing tumor growth and invasiveness. The aim of this study was to prepare a pair of monoclonal antibodys (mAbs) against GP73 and develop a newly designed double-antibody sandwich enzyme-linked immunosorbent assay (s-ELISA), which would be used in the detection of serum GP73 (sGP73) as well as in the diagnosis of HCC. Materials and Methods: Produced by prokaryotic expression, the purified recombinant GP73 (rGP73), produced by prokaryotic expression, was used to immunize the Balb/c mice. Two hybridoma cell lines against GP73 were obtained by fusing mouse Sp2/0 myeloma cells with spleen cells from the immunized mice. The titers of anti-GP73 mAb reached 1:243,000. Western blotting analysis and Immunohistochemistry staining revealed that anti-GP73 mAb could recognize GP73 protein. The double-antibody s-ELISA was successfully established and validated by 119 HCC and 103 normal serum samples. Results: showed that the detection limit of this method could reach 1.56 ng/ml, and sGP73 levels in HCC group (mean=190.6 ng/ml) were much higher than those of in healthy controls (mean=70.92 ng/ml). Conclusions: Results of our study not only showed that sGP73 levels of HCC patients were significantly higher than those of healthy controls, but also indicated that the laboratory homemade anti-GP73 mAbs could be the optimal tool used in evaluating sGP73 levels, which would provide a solid foundation for subsequent clinical applications.

Comparison of Biological Activities of Epimedium koreanum Nakai Produced in Korea and China (국내산과 중국산 삼지구엽초의 생리활성 비교)

  • Noh, Joon-Hyun;Kim, Young-Jin;Kim, Se-Won;Lee, Jin-Ha;Lee, Hyeon-Yong
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.3
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    • pp.195-200
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    • 2003
  • This study was carried out to compare the biological activities of Epjmedium koreanum Nakai grown wild in Korea and China. The antioxidative effect of E. koreanum Nakai-extracts grown wild in Gangwondo was 78%, which was higher than that in China as 71%. The inhibition ratio of growing cancer cells was estimated as 89, 91 and 86% for human liver, lung and breast cancer cell lines, respectively in adding 0.1 g/l of ethanol extracts of E. koreanum grown in Korea. The hepatoprotective effect and Angiotensin Converting Enzyme (ACE) inhibiting effect of Korean one were also observed as 124 and 87%, respectively, which were also higher than those produced in China. There was not much difference between Korean and Chinese one in inhibiting ${\alpha}-glucosidase$ activities in all ranges of supplement. It was proved that most effective extraction solvent was mixed type as water and ethanol (1:1, v/v) to have higher biological activities from both Korean and Chinese ones.

Redox Factor-1 Inhibits Cyclooxygenase-2 Expression via Inhibiting of p38 MAPK in the A549 Cells

  • Yoo, Dae-Goon;Kim, Cuk-Seong;Lee, Sang-Ki;Kim, Hyo-Shin;Cho, Eun-Jung;Park, Myoung-Soo;Lee, Sang-Do;Park, Jin-Bong;Jeon, Byeong-Hwa
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.3
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    • pp.139-144
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    • 2010
  • In this study, we evaluated the role of apurinic/apyrimidinic endonuclease1/redox factor-1 (Ref-1) on the tumor necrosis factor-$\alpha$ (TNF-$\alpha$) induced cyclooxygenase-2 (COX-2) expression using A549 lung adenocarcinoma cells. TNF-$\alpha$ induced the expression of COX-2 in A549 cells, but did not induce BEAS-2B expression. The expression of COX-2 in A549 cells was TNF-$\alpha$ dose-dependent (5~100 ng/ml). TNF-$\alpha$-stimulated A549 cells evidenced increased Ref-1 expression in a dose-dependent manner. The adenoviral transfection of cells with AdRef-1 inhibited TNF-$\alpha$-induced COX-2 expression relative to that seen in the control cells ($Ad{\beta}gal$). Pretreatment with $10\;{\mu}M$ of SB203580 suppressed TNF-$\alpha$-induced COX-2 expression, thereby suggesting that p38 MAPK might be involved in COX-2 expression in A549 cells. The phosphorylation of p38 MAPK was increased significantly after 5 minutes of treatment with TNF-$\alpha$, reaching a maximum level at 10 min which persisted for up to 60 min. However, p38MAPK phosphorylation was markedly suppressed in the Ref-1-overexpressed A549 cells. Taken together, our results appear to indicate that Ref-1 negatively regulates COX-2 expression in response to cytokine stimulation via the inhibition of p38 MAPK phosphorylation. In the lung cancer cell lines, Ref-1 may be involved as an important negative regulator of inflammatory gene expression.

Methylated Alteration of SHP1 Complements Mutation of JAK2 Tyrosine Kinase in Patients with Myeloproliferative Neoplasm

  • Yang, Jun-Jun;Chen, Hui;Zheng, Xiao-Qun;Li, Hai-Ying;Wu, Jian-Bo;Tang, Li-Yuan;Gao, Shen-Meng
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.6
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    • pp.2219-2225
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    • 2015
  • SHP1 negatively regulates the Janus kinase 2/signal transducer and activator of transcription (JAK2/STAT) signaling pathway, which is constitutively activated in myeloproliferative neoplasms (MPNs) and leukemia. Promoter hypermethylation resulting in epigenetic inactivation of SHP1 has been reported in myelomas, leukemias and other cancers. However, whether SHP1 hypermethylation occurs in MPNs, especially in Chinese patients, has remained unclear. Here, we report that aberrant hypermethylation of SHP1 was observed in several leukemic cell lines and bone marrow mononuclear cells from MPN patients. About 51 of 118 (43.2%) MPN patients including 23 of 50 (46%) polycythaemia vera patients, 20 of 50 (40%) essential thrombocythaemia and 8 of 18 (44.4%) idiopathic myelofibrosis showed hypermethylation by methylation-specific polymerase chain reaction. However, SHP1 methylation was not measured in 20 healthy volunteers. Hypermethylation of SHP1 was found in MPN patients with both positive (34/81, 42%) and negative (17/37, 45.9%) JAK2V617F mutation. The levels of SHP1 mRNA were significantly lower in hypermethylated samples than unmethylated samples, suggesting SHP1 may be epigenetically inactivated in MPN patients. Furthermore, treatment with 5-aza-2'-deoxycytidine (AZA) in K562 cells showing hypermethylation of SHP1 led to progressive demethylation of SHP1, with consequently increased reexpression of SHP1. Meanwhile, phosphorylated JAK2 and STAT3 were progressively reduced. Finally, AZA increased the expression of SHP1 in primary MPN cells with hypermethylation of SHP1. Therefore, our data suggest that epigenetic inactivation of SHP1 contributes to the constitutive activation of JAK2/STAT signaling. Restoration of SHP1 expression by AZA may contribute to clinical treatment for MPN patients.

Effects of Artemisia capillaris Methanol Extract on CD3+, CD4+, CD8+ and TNF-${\alpha}+$ Splenic Cells in Tumor Cells Inoculated Mice (종양 유발 마우스의 CD3+, CD4+, CD8+ 및 TNF-${\alpha}+$ 비장세포에 인진쑥 methanol 추출물이 미치는 영향)

  • Kim, Hong-Tae;Ku, Sae-Kwang;Kim, Ju-Wan;Jin, Tae-Won;Lim, Mee-Kyung;Kim, Ji-Eun;Do, Yoon-Jung;Yeo, Sang-Geon;Jang, Kwang-Ho;Oh, Tae-Ho;Lee, Keun-Woo
    • Journal of Veterinary Clinics
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    • v.26 no.1
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    • pp.1-7
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    • 2009
  • The Artemisia capillaris THUNB is a perennial herb that belongs to the family Compositae spp and probably the most common plant among the various herbal folk remedies being used in the treatment of abdominal pain, hepatitis, chronic liver disease, jaundice and coughing in Korea. Recently the biological and pharmacological actions of herb have been studied well such as antibacterial, antidiabetic and antitumor activities. This experiment was conducted to investigate antitumor and immunomodulatory effects of Artemisia capillaris extracts against Hepa-1c1c7 and Sarcoma 180 cancer cells in in vivo experimental tests. In in vivo experimental tests using 210 ICR mice, based on flow cytometry, CD3+, CD4+, CD8+ and TNF-${\alpha}+$ splenocytes were significantly (p<0.05) reduced in the Hepa-1c1c7 and Sarcoma 180 inoculated vehicle controls, HP and SP, compared to those of the intact vehicle control on both the $28^{th}$ day and the $42^{nd}$ day, respectively. These decreases of CD3+, CD4+, CD8+ and TNF-${\alpha}+$ cells induced by tumor inoculations were significantly (p<0.05) inhibited by mACH treatment regardless of the type of experiments and tumor cells inoculated. The results suggest that Artemisia capillaris methanol extracts have prominent antitumor effects on the cancer cell lines Hepa-1c1c7 and Sarcoma 180.

Antitumor and Immuno-potentiating Activity against Mouse Sarcoma 180 by Crude Polysaccharides Extracted from Fruiting Body of Tricholoma matsutake (송이(Tricholoma matsutake)의 자실체에서 추출한 조다당류가 생쥐의 Sarcoma 180에 미치는 항암 및 면역증강 작용)

  • Hur, Hyun;Choi, Yon-Il;Lee, Tae-Soo
    • Journal of Life Science
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    • v.18 no.9
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    • pp.1290-1298
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    • 2008
  • Tricholoma matsutake, one of edible and medicinal mushroom belonging to Tricholomaceae of Agaricales, has been known to contain some curing effect on gastric cancer and ulcer, and inhibitory effect on sarcoma 180 and Ehrlich sarcoma. Neutral salt soluble (0.9% NaCl), hot water soluble and methanol soluble substances (hereinafter referred to Fr. NaCl, Fr. HW and Fr. MeOH, respectively) were extracted from fruiting body of the mushroom. In vitro cytotoxicity tests, crude polysaccharides were not cytotoxic against cancer cell lines such as Sarcoma 180, HepG2, HT29 and NIH3T3 at the concentration of 2.0 mg/ml. Intraperitoneal injection with crude polysaccharides showed life prolongation effect of 23.4$\sim$37.2% in mice previously inoculated with Sarcoma 180. Fr. MeOH and Fr. HW exhibited the immuno-potentiating activity of B lymphocyte by increasing the alkaline phosphatase activity by 2.2$\sim$11.9 folds compared with control at the concentration of 0.2$\sim$0.5 mg/ml. In case of Fr. NaCl, the numbers of peritoneal exudate cells and circulating leukocytes were increased by 6.0 and 1.5 folds at the concentration of 50 mg/kg, respectively. Therefore, it is concluded that crude polysaccharides extracted from fruiting body of Tricholoma matsutake showed antitumor and immuno-potentiating activity against Sarcoma 180 of mouse.

Nutritional Component and Anticancer Properties of Various Extracts from Haesongi Mushroom (Hypsizigus marmoreus) (해송이버섯(Hypsizigus marmoreus)의 영양성분과 추출용매에 따른 암세포 생장억제 효과)

  • Jung, Eun-Bong;Jo, Jin-Ho;Cho, Seung-Mock
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.11
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    • pp.1395-1400
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    • 2008
  • This study was aimed to analyze the nutritional components and anticancer properties of Haesongi mushroom (Hypsizigus marmoreus), which has been recently available in Korea, to estimate its nutritional and functional values. Fruit body of Haesongi mushroom was investigated for its proximate components and mineral contents. Its water and ethanol extracts were compared for nutritional components such as $\beta$-glucan, protein, and total sugar. Anticancer effects of both extracts were measured against human cancer cell lines in vitro. This mushroom contained high protein (22.63%), total dietary fiber (30.80%), and K (3383.3 mg/100 g). The water extract contained more nutritional components such as $\beta$-glucan (9.32 mg/g), protein (17.71%), and total sugar (39.93%), compared with the ethanol extract. Moreover the extraction yield of the water extract was higher than the ethanol extract. The growth inhibitory effects of the water extract (5 mg/mL) on AGS, HepG2, and SW480 human cancer cells were 90.61, 75.43, and 58.49%, respectively. However, the ethanol extract showed 81.79, 49.90, and 25.71% growth inhibition, respectively. In this study, it is demonstrated that water is a more efficient solvent than ethanol for extracting nutritional and functional components from Haesongi mushroom.

Effects of Artemisia capillaris Methanol Extract on the Amounts of Splenocytes-derived Cytokines in Tumor Cells Inoculated Mice (인진쑥 Methanol 추출물이 암이 유발된 마우스의 비장세포 유래 Cytokine 함량에 미치는 영향)

  • Kim, Hong-Tae;Ku, Sae-Kwang;Kim, Ju-Wan;Jin, Tae-Won;Koo, Sung-Wook;Lim, Mee-Kyung;Do, Yoon-Jung;Jang, Kwang-Ho;Oh, Tae-Ho;Lee, Keun-Woo
    • Journal of Veterinary Clinics
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    • v.26 no.5
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    • pp.408-412
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    • 2009
  • The Artemisia capillaris THUNB is a perennial herb that belongs to the family Compositae spp. and probably the most common plant among the various herbal folk remedies being used in the treatment of abdominal pain, hepatitis, chronic liver disease, jaundice and coughing in Korea. This experiment was conducted to investigate the effects of Artemisia capillaris extracts on the amounts of splenocytes-derived cytokine ($TNF-{\alpha},\;IL-1{\beta}$ and IL-10). In in vivo experimental tests using 210 ICR mice with Hepa-1c1c7 or sarcoma 180 cancer line, splenocytes derived cytokine contents were significantly (p < 0.05) reduced in the Hepa-1c1c7 and Sarcoma 180 inoculated vehicle controls, HP and SP, compared to those of the intact vehicle control on both the $28^{th}$ day and the $42^{nd}$ day, respectively. However, these decreases of $TNF-{\alpha},\;IL-1{\beta}$ and IL-10 levels induced by tumor inoculations were significantly (p < 0.01, p < 0.05) inhibited by mACH (Artemisia capillaris methanol extracts) treatment regardless of the type of experiments and tumor cells inoculated. The results suggest that Artemisia capillaris methanol extracts have prominent anti-inflammation effects on the cancer cell lines Hepa-1c1c7 and Sarcoma 180.