• Title/Summary/Keyword: human liver cell line

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Identification of Differentially Expressed Genes (DEGs) by Malachite Green in HepG2 Cells

  • Kim, Youn-Jung;Song, Mee;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.4 no.1
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    • pp.22-30
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    • 2008
  • Malachite Green (MG), a toxic chemical used as a dye, topical antiseptic and antifungal agent for fish, is highly soluble in water, cytotoxic to various mammalian cells and also acts as a liver tumor promoter. In view of its industrial importance and possible exposure to human beings, MG possesses a potential environmental health hazard. So, we performed with HepG2, a human hepatocellular carcinoma cell line, to identify the differentially expressed genes (DEGs) related to toxicity of MG. And we compared gene expression between control and MG treatment to identify genes that are specifically or predominantly expressed by employing annealing control primer (ACP)-based $GeneFishing^{TM}$ method. The cytotoxicity $(IC_{20})$ of MG was determined above the $0.867{\mu}M$ in HepG2 cell for 48 h treatment. And the DEGs of MG were identified that 5 out of 6 DEGs were upregulated and 1 out of 6 DEGs was down-regulated by MG. Also, MG induced late apoptosis and necrosis in a dose dependent in flow cytometric analysis. Through further investigation, we will identify more meaningful and useful DEGs on MG, and then can get the information on mechanism and pathway associated with toxicity of MG.

Effect of Fermented Ginseng Extract by Mushroom Mycelia on Antiproliferation of Cancer Cells (버섯균사체로 발효시킨 인삼 추출물의 암세포 증식억제 효과)

  • Kim, Hyun-Young;Joung, Eun-Mi;Hwang, In-Guk;Jeong, Jae-Hyun;Yu, Kwang-Won;Lee, Jun-Soo;Jeong, Heon-Sang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.1
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    • pp.36-41
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    • 2010
  • This study was conducted to investigate the effects of fermented ginseng extract by mushroom mycelia on antiproliferation of cancer cells. Phellinus linteus, Ganoderma lucidum, and Hericium erinaceum mycelia were inoculated to ginseng. The effects of fermented ginseng extract on antiproliferation of stomach (MKN-45), colon (HCT116), mammary (MCF-7), lung (NCIH460), prostate (PC-3), and liver (HepG2) cancer cells were investigated by MTT assay. Fermented ginseng extract showed significant antiproliferation effects compared with fresh ginseng extract. Fermented ginseng extract by P. linteus, G. lucidum, and H. erinaceum mycelia showed growth-inhibitory effect of 44.50, 17.75 and 43.98% viability at 1.5 mg/mL on the MKN-45 cell line, 62.86, 3.73, and 54.55% at 1.5 mg/mL on the HCT116 cell line, 41.81, 7.01, and 37.84% at 1.5 mg/mL on the MCF-7 cell line, 53.52, 5.31, and 35.27% at 1.5 mg/mL on the NCIH460 cell line, 35.05, 3.07, and 44.29% at 1.5 mg/mL on the PC-3 cell line, and 59.57, 6.34, and 4.97% at 1.5 mg/mL on the HepG2 cell line, respectively. These results indicated that fermented ginseng by G. lucidum mycelium showed the highest antiproliferation effect against various cancer cells.

Analysis for Compatibility of Gyejibongnyeong-hwan and Its Biological Activities (계지복령환(桂枝茯苓丸)의 배오(配伍)분석과 그 생리활성)

  • Kim, Do Hoy;Jung, Yang Sam;Yoon, Michung;Yoon, Yoosik;Shin, Soon Shik
    • Herbal Formula Science
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    • v.24 no.4
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    • pp.353-365
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    • 2016
  • Objectives : We analysed Gyejibongnyeong-hwan's compatibility principle and investigated biological activities by categorizing with molecular level, cellular level, animal level and human level based on Korean study for this formula. Methods : Gyejibongnyeong-hwan's compatibiltity principle was examined by the system of chief, deputy, assistant, and envoy. We looked into studies that presented in Korea from 1956 to 2016 about Gyejibongnyeong-hwan through Korea Institute of Oriental Medicine, Korean medicine information system (OASIS). Then classify into molecular level, cellular level, animal level and human level to analyse. Results : According to the system of chief, deputy, assistant, and envoy, chief herb is Cinnamomi Ramulus, deputy herb is Persicae Semen, assistant herb is Moutan Cortex, Paeoniae Radix, Poria, and envoy herb is Mel. Biological activities can be detected in transcription factors, enzymes, and inflammatory mediators for molecular level. For cellular level, it can be determined in human uterine endometrial cancer cell, human hepatocarcinoma cell, and human platelets. In mouse and rats for animal level, in overian cystoma, menorrhalgia, quality of life improvement in postmenopausal women, and blood stasis with motor vehicle accident for human level, biological activities was caught. Conclusions : From above results, Gyejibongnyeong-hwan is composed in line with the system of chief, deputy, assistan, and envoy. Biological activities are effective to improvement of menorrhalgia, anti-cancer, anti-oxidative, anti-inflammation, improvement of atherosclerosis, analgesic, anti-convulsion, wound healing, and improvement of liver function.

Protective effects against alcoholic liver damage: potential of herbal juice (HJ), blend of Zingiber officinale Roscoe and Pueraria lobata Ohwi extracts

  • Young Yun Jung;You Yeon Choi;Woong Mo Yang;Kwang Seok Ahn
    • Journal of Convergence Korean Medicine
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    • v.5 no.1
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    • pp.45-54
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    • 2023
  • Objectives : Alcohol-induced liver disease advances as to reactive oxygen species (ROS) and cellular lipid peroxidation increase. We examined the hepatoprotective effects of Zingiber officinale Roscoe rhizome extract (ZR), Pueraria lobata Ohwi flower extracts (PF), and a newly developed herbal juice (HJ), which was a combination of ZR and PF extracts, against ethanol-induced hepatotoxicity. Methods: The study utilized the human hepatoma cell line HepG2 cells to validate the hepatoprotective effect of HJ (50~200 ㎍/mL) against ethanol (EtOH, 700 mM)-induced liver damage. Results: HJ effectively reduced the protein expression of sterol regulatory element-binding transcription factor 1, adiponectin, and AMP-activated protein kinase in EtOH-induced HepG2 cells. The levels of ROS, total cholesterol, and triglycerides, which are the result of various synthesis and lipogenesis processes induced by EtOH in the liver, were reduced by HJ. Furthermore, the activities of alcohol dehydrogenase and aldehyde dehydrogenase, enzymes linked to alcohol degradation, were more effectively downregulated by HJ treatment compared to treatment with ZR and PF alone, all without causing cytotoxic effects. Conclusions: HJ protects the liver by inhibiting EtOH-induced lipogenesis, lowering ROS generation, and improving alcohol degradation, which is more effective than ZR and PF alone. Further, in vivo experiments can offer additional evidence regarding the effectiveness, safety, and underlying mechanism of action of HJ.

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A study on separation and characterization of matrix metalloproteinase-9 inhibitors from natural plants (천연 식물 추출물에서 Matrix Metalloproteinase-9 활성 억제제의 분리 및 특성화에 관한 연구)

  • Hur, Yong-Chul;Park, Sung-Woo;Kim, Tai-Jin
    • Analytical Science and Technology
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    • v.18 no.3
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    • pp.188-193
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    • 2005
  • Three different oriental natural plants (Angelicae Dahuricae Radix, Sanguisorba Officinalis L, Euonymus alatus) were extracted with 70% methanol under refluxing for 4 hr in order to investigate their inhibitory effect on Matrix Metalloproteinase-9 (MMP-9) by a modified gelatin zymography, where only euonymus alatus showed the inhibitory effect on the activity of Matrix MMP-9. The fraction was collected by using the mixture of ethyl acetate and hexane on silica gel column. Seven portions were obtained and three fractions of them (first, third and forth) showed inhibitory effect on the zymography. To verify the effect of these substances on cells, human hepatoma, Hep3B cells as a cancer model, and Chang liver cells as a normal model were selected. In order to examine the cell viability, $1{\mu}g/mL$ of each extract was treated on cells. Most of the methanol soluble fractions showed negligible toxicity on human liver cell line.

Study on the Anti-cancer, Anti-metastasis and Immune response improvement of Aqua-acupuncture with Cistanches Herba infusion solution (육종용약침의 항암작용(抗癌作用) 및 면역효과(免疫效果)에 대(對)한 실험적(實驗的) 연구(硏究))

  • Lee, Eun-Yong;Lee, Byung-Ryul
    • Journal of Acupuncture Research
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    • v.17 no.1
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    • pp.251-286
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    • 2000
  • To study the effects of anti-cancer, anti-metastasis and immune response improvement of aqua-acupuncture with Cistanches Herba infusion solution, we used Cistanches Herba infusion solution(taken by water-alcohol method) put into Chung-wan(CV12) and Chok-Samni(S36) of BALB/c or C57BL/6 which are corresponding to human body. We observed the cytotoxicity, the effect on the expression of MMP-9 gene, the ability to control cancer cell proliferation, change of body weight, surviving number, MST, ILS, changes in amount of WBC, RBC, PLT, total protein, creatinine, glucose and LDH, weight of spleen, number of pulmonary colony, histological analysis on tissue metastasis of lung and liver, splenic cell proliferation, the expression of cytokine gene, the number of $CD4^+$, $CD8^+$, $CD19^+$ and NK cell, and concluded like this. The results were obtained as follows : 1. The cytotoxicity about B16-F10 cell line of $2^0$, $2^{-1}$, $2^{-2}$, $2^{-5}$ diluent groups in Cistanches Herba infusion solution treatment was inhibited significantly, compared with control group. 2. The cytotoxicity about HT1080 cell line of $2^0$, $2^{-1}$, $2^{-2}$, $2^{-3}$, $2^{-5}$, $2^{-8}$ diluent groups in Cistanches Herba infusion solution treatment was inhibited significantly, compared with control group. 3. The effect on expression of MMP-9 gene was decreased in all the sample groups, compared with control group. 4. The effect on the control-ability on the cancer cell proliferation showed cytotooicity significantly in $2^0$, $2^{-1}$, $2^{-2}$, $2^{-3}$, $2^{-4}$, $2^{-5}$ diluent groups. 5. S-180 cancer cell line transplants in BALB/c mice were inhibited significantly in weight increase in all the sample groups, compared with control group. The surviving number increased in almost sample groups, except one group put into Chok-Samni(S36) with 20% Cistanches Herba infusion solution treatment group that showed same number of the control group. 6. S-180 cancer cell line transplants in BALB/c mice showed high MST and ILS significantly in almost sample groups, compared with control group. But one group put into Chok-Samni(S36) with 20% Cistanches Herba infusion solution treatment group showed low MST and ILS than control group. 7. The sample group injected in vein with B16-F10 cancer cell line in C57BL/6 mice showed increased ILS compared with control group significantly in anti-metastasis test. 8. The sample group injected in vein with B16-F10 cancer cell line in C57BL/6 mice were increased significantly in the number of WBC and glucose, and decreased significantly in the amount of LDH, compared with control group. However, there's no significant increase or decrease in number of RBC, PLT, total protein and creatinine. 9. We couldn't find any significant relation in spleen weight of the sample group. 10. In pulmonary colony, sample group was decreased significantly, compared with control group. 11. Histological analysis of sample group inhivited compared with that of control group in both of lung and liver. 12. In immune system, all the sample groups showed having more relevancy to the effect on splenic cell proliferation than normal group. 13. The effect on cytokine gene expression of all the sample groups were increased than control group. 14. In flow cytometry there's no significant relation in number of $CD8^+$, $CD19^+$ cell, however, the number of $CD4^+$ cell and NK cell in sample groups were increased than in control group. Above the results showed that aqua-acupuncture of Cistanches Herba infusion solution has effects of anti-cancer, anti-metastasis and immune response improvement.

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Effects of Loquat (Eriobotrya japonica Lindl.) Ethanol Extracts of Different Aerial Parts on Antioxidant Activity and Antiproliferation of Human Cancer Cells (비파 부위별 에탄올 추출물의 항산화 활성 및 암세포 증식 억제효과)

  • Lee, Hwan;Kim, Yeon-Kyoung;Lee, Hyun-Joo;Lee, Jae-Joon
    • The Korean Journal of Community Living Science
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    • v.27 no.2
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    • pp.211-220
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    • 2016
  • The current study was carried out to determine the effects of the seed, flesh (seedless fruit), and leaf of loquat (Eriobotrya japonica Lindle.) on antioxidative activity and anti-proliferation in human cancer cells. Total polyphenol contents of loquat seed, flesh, and leaf ethanol extracts were found to be 17.77, 32.32, and 28.08 mg/g, respectively. Also, total flavonoid contents of loquat seed, flesh, and leaf ethanol extracts were found to be 18.77, 28.73, and 21.35 mg/g, respectively. The $IC_{50}$ values of DPPH hydroxyl scavenging of loquat seed, flesh, and leaf ethanol extracts were 0.049, 0.063, and 0.042 mg/mL, respectively. Antioxidative indexes of loquat leaf and seed ethanol extracts was highly and it was similar to the BHA and BHT. The antioxidative activities in loquat seed and leaf ethanol extracts were higher in loquat flesh. The antiproliferation effect of loquat seed and leaf ethanol extracts on liver cancer cell line (H460), stomach cancer cell line (AGS) and lung cancer cell line (A549) showed higher values compared with the flesh ethanol extracts. These results indicate that loquat ethanol extracts may play a positive role in antioxidative properties and cancer prevention.

Antioxidant and Cell Proliferation Effects of Acanthopanax senticosus for Extract in Human Osteoblast-like MG-63 Cell Line (가시오가피 추출물의 항산화활성 및 MG-63 조골세포 증식과 alkaline phosphatase 활성에 미치는 효과)

  • Lim, So-Young;Leem, Jae-Yoon;Lee, Choong-Soo;Jang, Yu-Jung;Park, Jeong-Woo;Yoon, Sun
    • Korean Journal of Food Science and Technology
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    • v.39 no.6
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    • pp.694-700
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    • 2007
  • Acanthopanax senticosus is a common Asian herb also known as "Siberian Ginseng". It is often used as a traditional herbal medicine for reducing damage in the liver, kidney, bone and muscle. In the present study we investigated the ferric reducing/antioxidant power and total phenolic contents of the ethanol-/water-extracts obtained from the stems and leaves of Acanthopanax senticosus. Osteoblast cellular proliferation was evaluated using the MTT and alkaline phosphatase activity assays in the human osteoblast-like MG-63 cell line. Acanthopanax senticosus extracts exerted remarkable ferric reducing/antioxidant power and contained high amount of phenolics. Among the extracts the stem-/ethanol-extract showed the highest antioxidant activity and total phenol content. Interestingly a highly positive correlation was found between antioxidant activity and total phenol content (p < 0.01). Proliferation of MG-63 osteoblast cells was highest in the stem-/ethanol-extract and alkaline phosphatase activity significantly increased in the water-extract of the stems (p < 0.05). These findings suggest that Acanthopanax senticosus extracts have antioxidant activity for preventing oxidative stress-related diseases and may have beneficial effects on bone health through the proliferation of osteoblast cells.

Effects of Epothilone A in Combination with the Antidiabetic Drugs Metformin and Sitagliptin in HepG2 Human Hepatocellular Cancer Cells: Role of Transcriptional Factors NF-κB and p53

  • Rogalska, Aneta;Sliwinska, Agnieszka;Kasznicki, Jacek;Drzewoski, Jozef;Marczak, Agnieszka
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.3
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    • pp.993-1001
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    • 2016
  • Type 2 diabetes mellitus patients are at increased risk of many forms of malignancies, especially of the pancreas, colon and hepatocellular cancer. Unfortunately, little is known of the possible interaction between antidiabetic drugs and anticancer agents. The present study investigates the influence of metformin (MET) and sitagliptin (SITA) on the in vitro anticancer activity of the microtubule depolymerization inhibitor agent epothilone A (EpoA). Hepatocellular liver carcinoma cell line (HepG2) viability and apoptosis were determined by the MTT test and by double staining with PO-PRO-1 and 7-aminoactinomycin D, respectively, after treatment with EpoA, metformin or sitagliptin. The levels of nuclear factor NF-${\kappa}B$ and p53 were evaluated in the presence and absence of inhibitors. While EpoA and MET inhibited HepG2 cell proliferation, SITA did not. EpoA and SITA induced higher p53 levels than MET. All tested drugs increased the level of NF-${\kappa}B$. Only MET enhanced the proapoptotic effect of EpoA. The EpoA+MET combination evoked the highest cytotoxic effect on HepG2 cells and led to apoptosis independent of p53, decreasing the level of NF-${\kappa}B$. These findings support the link between NF-${\kappa}B$ and p53 in the modulation of apoptotic effects in HepG2 cells treated by EpoA. Our studies indicate that the combination of EpoA and MET applied in subtoxic doses has a stronger cytotoxic effect on liver cancer cells than each of the compounds alone. The therapeutic advantages of the combination of EpoA with MET may be valuable in the treatment of patients with diabetes mellitus type 2 (T2DM) and liver cancer.

Effects of KHchunggan-tang on the Nonalcoholic Fatty Liver Disease in Palmitate-induced Cellular Model (Palmitate로 유발된 비알코올성 지방간 모델에 대한 KH청간탕(淸肝湯)의 효과 연구)

  • Han, Chang-Woo;Lee, Jang-Hoon
    • The Journal of Korean Medicine
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    • v.32 no.1
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    • pp.109-120
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    • 2011
  • Objectives: The aim of this investigation was to evaluate the efficacy of KHchunggan-tang aqueous extract on the experimental nonalcoholic fatty liver disease(NAFLD) induced by palmitate. Materials and Methods: To generate a cellular model of NAFLD, we used HepG2 cells, a human hepatoma cell line, treated with 0.5 mM palmitate. By this cellular model, effects of KHchunggan-tang aqueous extract were evaluated. Intracellular lipid accumulation, free radical formation, and apoptosis were detected by Nile red staining, 2',7'-dichloroflourescin diacetate(H2DCF-DA), and 4',6-diamidino-2-phenylindole(DAPI)/propidium iodide(PI) staining, respectively. Some proteins related with NAFLD were determined by western blot. Results: Typical pathological features of NAFLD occurred in the cellular model. Palmitate increased the levels of intracellular lipid vacuoles, decreased cell viability, and increased apoptosis. Palmitate increased free radical formation and lipid peroxidation, too. However, KHchunggan-tang aqueous extract reduced palmitate-induced pathologic features, i.e. steatosis, free radical formation, and apoptosis. In addition, KHchunggan-tang aqueous extract suppressed palmitate-activated c-Jun N-terminal kinase(JNK) signaling, and SP600125, a JNK inhibitor, significantly reversed the palmitate-induced pathologic changes as KHchunggan-tang aqueous extract. It means that the signaling pathway other than JNK can be involved in the KHchunggan-tang mediated cellular protection of palmitate-treated Hep G2 cells. Conclusions: These results suggest that KHchunggan-tang aqueous extract has hepatoprotective effects on NAFLD with combined properties in cellular steatosis, ROS production, and cytoprotection, and thus may have valuable clinical applications for treatment of this chronic liver disease.