• The Korea Journal of Herbology
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• v.21 no.4
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• pp.27-36
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• 2006

Objectives : This study was performed for the investigation of anticancer effects of methanol extract of Rheum Rhizoma (MeOH-RR) on a human liver cancer cell line (HepG2). Methods : To study the cytotoxic effect of MeOH-RR on HepG2 cells, the cell viability was determined by XTT reduction method and trypan blue exclusion assay. The cleavage of poly ADP-ribose polymerase (PARP), a substrate for caspase-3 and a typical sign of apoptosis, and the activation of procaspase-3, -8 and -9 were examined by western blot analysis. Furthermore, MeOH-RR-induced apoptosis was confirmed by DNA fragmentation. The release of cytochrome c from mitochondria to cytosol, the level of Bcl-2 and Bax were examined by western blot analysis. Results : MeOH-RR reduced proliferation of HepG2 cells in a dose-dependent manner at 24 h and 48 h treatment. MeOH-RR induced the activation of caspase-3, -8, and -9 and the cleavage of poly ADP-ribose polymerase (PARP), a substrate for caspase-3. Furthermore, treatment with MeOH-RR resulted in internucleosomal DNA fragmentation, evidenced by the formation of a DNA ladder on agarose gel, a hallmark of cells undergoing apoptosis. MeOH-RR downregulated Bcl-2, upregulated Bax, and increased the release of cytochrome c from the mitochondria into cytosol in a dose-dependent manner. Moreover, MeOH-RP increased caspase-3 activity. Conclusion : There results suggest that MeOH-RR induce apoptosis via mitochondrial pathway and caspase-3-dependent pathway in HepG2 cells. There results suggest that MeOH-RR is potentially useful as a chemotherapeutic agent in human liver cancer.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.11
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• pp.6363-6367
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• 2013

Atractylis lancea (Thunb.) DC. (AL), an important medicinal herb in Asia, has been shown to have anti-tumor effects on cancer cells, but the involved mechanisms are poorly understood. This study focused on potential effects and molecular mechanisms of AL on the proliferation of the Hep-G2 liver cancer cell line in vitro. Cell viability was assessed by MTT test in Hep-G2 cells incubated with an ethanol extract of AL. Then, the effects of AL on apoptosis and cell cycle progression were determined by flow cytometry. Telomeric repeat amplification protocol (TRAP) assays was performed to investigate telomerase activity. The mRNA and protein expression of human telomerase reverse transcriptase (hTERT) and c-myc were determined by real-time RT-PCR and Western blotting. Our results show that AL effectively inhibits proliferation in Hep-G2 cells in a concentrationand time-dependent manner. When Hep-G2 cells were treated with AL after 48h,the $IC_{50}$ was about 72.1 ${\mu}g/mL$. Apoptosis was induced by AL via arresting the cells in the G1 phase. Furthermore, AL effectively reduced telomerase activity through inhibition of mRNA and protein expression of hTERT and c-myc. Hence, these data demonstrate that AL exerts anti-proliferative effects in Hep-G2 cells via down-regulation of the c-myc/hTERT/telomerase pathway.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.7
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• pp.3123-3128
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• 2014

The liver is normally the major site of glucose metabolism in intact organisms and the most important target organ for the action of insulin. It has been widely accepted that insulin resistance (IR) is closely associated with postoperative recurrence of hepatocellular carcinoma (HCC). However, the relationship between IR and drug resistance in liver cancer cells is unclear. In the present study, IR was induced in HepG2 cells via incubation with a high concentration of insulin. Once the insulin-resistant cell line was established, the stability of HepG2/IR cells was further tested via incubation in insulin-free medium for another 72h. Afterwards, the biological effects of insulin resistance on adhesion, migration, invasion and sensitivity to cis-platinum (DDP) of cells were determined. The results indicated that glucose consumption was reduced in insulin-resistant cells. In addition, the expression of the insulin receptor and glucose transportor-2 was downregulated. Furthermore, HepG2/IR cells displayed markedly enhanced adhesion, migration, and invasion. Most importantly, these cells exhibited a lower sensitivity to DDP. By contrast, HepG2/IR cells exhibited decreased adhesion and invasion after treatment with the insulin sensitizer pioglitazone hydrochloride. The results suggest that IR is closely related to drug resistance as well as adhesion, migration, and invasion in HepG2 cells. These findings may help explain the clinical observation of limited efficacy for chemotherapy on a background of IR, which promotes the invasion and migration of cancer cells.

• Journal of Nutrition and Health
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• v.35 no.4
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• pp.439-445
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• 2002

This study was undertaken to investigate the inhibitory effect of prunus fume extracts on the growth of Hep G2 and HeLa cells. Prunus mums was extracted using the following solvents hexane, chloroform, ethylacetate, methanol, and hot water. The effect on the growth of each cancer cell line was examined by MTT (3-[4, 5-dimethylthiaeol-2-yl)-2,5-diphenyl tetrazolium bromide) assay, cytotoxicity testing, and microscopic observation. The ethylacetate extracts of Prunus muse at the concentration of 250 $\mu\textrm{g}$/ml exhibited the greatest inhibitory effect on the growth of Hep G2 in the MW assay. In cytotoxicity testing, the treatment of the Hep G2 cells with ethylacetate extracts (1000 $\mu\textrm{g}$/ml for 72 hrs) destroyed 75% of the cells, and morphological changes were also observed. futhermore, the hexane extracts of Prunes muse at the concentration of 250 $\mu\textrm{g}$/ml exhibited the greatest inhibitory effect on the growth of HeLa cells in the MTT assay. The treatment of the HeLa cells with the hexane extracts (1000 $\mu\textrm{g}$/ml for 72 hrs) resulted in the destruction of 68% of the cells. Fibroblasts were not affected by either ethylacetate or hexane extracts of prunus muse.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.12
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• pp.1779-1784
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• 2015

This study was performed to investigate the effects of various garlic extracts on cholesterol synthesis in HepG2 cells. Raw garlic, grilled garlic, and freeze dried garlic were subjected to cold water extraction, and extracts were incubated at room temperature for 1 min or 60 min. The extracts were treated to HepG2 cells for 4 h, and cholesterol synthesis and mRNA expression level of HMG-CoA reductase were investigated. The alliin contents were reduced when garlic was incubated at room temperature for 60 min. Raw garlic extracts showed lower intracellular cholesterol contents compared to that of the control group. However, raw garlic extracts incubated for 60 min showed no differences compared to the control group. Freeze-dried garlic extract showed minimum intracellular triglyceride and cholesterol contents. Relative mRNA expression level of HMG-CoA reductase, a rate-limiting enzyme of cholesterol synthesis, decreased in the garlic extracts. Compared with 60 min, garlic extracts incubated for 1 min showed a reduced level of HMG-CoA reductase mRNA expression. The freeze-dried garlic extract reduced mRNA expression level of HMG-CoA reductase in a dose-dependent manner in cells treated with 5% of 0.2, 0.5, 0.8, 1.0, and 1.5 mg/mL in medium, and the effect was maxed out at dose of 5% garlic extract at 1.0 mg/mL in medium.

• Korean Journal of Food Science and Technology
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• v.43 no.6
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• pp.787-790
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• 2011

The objective of this study was to investigate the anticancer effects of Acer tegmentosum M. extracts. HepG2 hepatocarcinoma cells were treated with ethanol, chloroform, ethylacetate, butanol, aqueous fraction and hot water extract. The antiproliferative effect was evaluated by trypan blue exclusion, MTT-based viability assay and morphology. The trypan blue test showed that anticancer effect of the A. tegmentosum M. extracts on HepG2 cells increased gradually in proportion to the increasing concentration of the fractions. The butanol fraction showed the highest anticancer activity against HepG2 cells (p<0.05). The MTT assay indicated that the growth inhibition by the butanol fraction was dose-dependent. These results suggest that A. tegmentosum M. has the potential to inhibit the growth of hepatocarcinoma cells.

• Journal of Life Science
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• v.20 no.6
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• pp.877-884
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• 2010

We investigated the growth inhibitory effect of internal organs of Aplysia kurodai (AK) on proliferation in cancer cell lines in vitro. The internal organs of AK were extracted with methanol (AKM), which were then further fractionated into four subfractions by using solvent partition method, resulting in hexane (AKMH), methanol (AKMM), butanol (AKMB), and aqueous (AKMA) soluble fractions. We determined the cytotoxic effect of these four fractions in four kinds of cancer cell lines - HepG2, MCF-7, HT29 and B16-F10 - by MTT assay. Among the four subfractions of AKM, AKMM showed the strongest cytotoxic effects on all cancer cell lines which were used. Morphological changes such as membrane shrinking and blebbing of cells were also observed in AKMM treatment in HepG2 cells. In addition, we also observed quinone reductase (QR) induced effect in the methanol layer (AKMM) of HepG2 cells. AKMM showed the highest induction activity of quinone reductase on HepG2 cells among the partition layers. The QR induced effect of AKMM was determined to be 2.4 at $100\;{\mu}g/ml$ level with a control value of 1.0. Although further studies are needed, the present work suggests that internal organs of Aplysia kurodai (AK) may be a chemopreventive agent for the treatment of human cells.

• Journal of Life Science
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• v.27 no.2
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• pp.233-240
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• 2017

Previously, we have shown that Schisandra chinensis (Turcz.) Baill. (S. chinensis) has a protective effect against endoplasmic reticulum (ER) stress-induced hepatic steatosis. Gomisin A is a bioactive phytoestrogen derived from S. chinensis. In the present study, the in vitro and in vivo effects of gomisin A on ER stress and hepatic steatosis were investigated. We quantified the expression of markers of ER stress, including glucose regulated protein 78 (GRP78), C/EBP homolog protein (CHOP), and X-box-binding protein-1 (XBP-1), in HepG2 cells treated with tunicamycin or palmitate. Tunicamycin treatment in HepG2 cells induced the expression of markers of ER stress, including GRP78, CHOP, and XBP-1c. However, treatment with gomisin A reduced the expression of markers of ER stress. These inhibitory effects were also observed in palmitate-incubated HepG2 cells. The in vivo inhibitory effects of gomisin A were assessed in mice injected with tunicamycin or fed with a high fat diet (HFD). Gomisin A reduced the expression of markers of ER stress and decreased triglyceride levels in the livers of mice after tunicamycin injection or HFD feeding. Furthermore, gomisin A decreased the expression of inflammatory genes in palmitate-incubated HepG2 cells and the liver of HFD-fed obese mice. These results suggest that gomisin A inhibits ER stress and ameliorates hepatic steatosis induced by ER stress.

• Korean Journal of Plant Resources
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• v.28 no.5
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• pp.635-640
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• 2015

Nonalcoholic fatty liver disease (NAFLD) is a kind of liver inflammation caused by an accumulation of fat in the liver. Patients with NAFLD have an increased risk to develop liver fibrosis, which leads to cirrhosis. To investigate hepatoprotective effects of Agrimonia eupatoria L (A. eupatoria), oleic acid-induced NAFLD in HepG2 cells was used and A. eupatoria was fractionated with ethanol (EtOH), n-hexane, dichloromethane (CH₂Cl₂), ethyl acetate (EtOAc), n-butanol (BuOH), and H₂O. Cells treated with the EtOAc fraction showed the highest lipid accumulation inhibiting effect. A. eupatoria also suppressed triglyceride accumulation and inhibited expression of lipid marker gene, such as a peroxisome proliferator activated receptor γ (PPAR-γ). Moreover, another marker, mRNA expression level of peroxisome proliferator activated receptor α (PPAR-α) was significantly increased by in a dose-dependent manner. These results suggest that A. eupatoria is a potent agent for the treatment of NAFLD.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.3
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• pp.288-293
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• 2008

In this study, we investigated the anticancer activity of Makgeoly (MG). MG was fractionated into four fractions by using solvent partition method, affording hexane (MGH), methanol (MGM), butanol (MGB) and aquous (MGA) soluble fractions. We determined the cytotoxicity of these four fractions in four kinds of cancer cell lines, such as HepG2, MCF-7, B16-F10 and HT29 by MTT assay. Among the various fractions, the MGM showed the strongest cytotoxic effects on all cancer cell lines. The morphological changes such as membrane shrinking and blebbing of cells were also observed by MGM treatment in HepG2 cell. In addition, we observed quinone reductase (QR) activity stimulating effects in all fraction layers of MG on HepG2 cells. QR activity increased approximately 2.6 and 2.1 times in MGM and MGH treated HepG2 cell at $100{\mu}g/mL$, respectively, compared to that in control value. Although further studies are needed, the present work could suggest that the fin of MG has a potential to be used as a chemopreventive agent against cancer.