• Applied Biological Chemistry
• /
• v.49 no.2
• /
• pp.91-96
• /
• 2006

Antioxidant activities and biological properties such as antimutagenic and cytotoxic effect of Phellinus linteus extracts from different extraction conditions were measured against Salmonella typhimurium and human cancer cell lines. DPPH free radical scavenging activities of the extracts were higher in the solutions extracted with ethanol (17.14) and ethanol after water (17.79), respectively. In the Ames test, ethanol extract of P. linteus alone did not exhibit any mutagenicity but showed substantial inhibitory effect against mutations induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), 4-nitroquinoline-1-oxide (4NQO), and benzo $({\alpha})$ pyrene $(B({\alpha})P)$. The extracts of ethanol and ethanol after water of P. linteus $(200\;{\mu}g/plate)$ had the highest inhibitory effect of 61.5 and 60.9%, respectively, on the mutagenesis on S. typhimurium TA98 strain induced by $B({\alpha})P$. Extracted solutions of ethanol and ethanol after water of P. linteus showed high antimutagenic effect against MNNG, 4NQO, Trp-P-1 and $B({\alpha})P$, causing mutations in S. typhimurium TA100 strain. The anticancer effects of P. linteus extracts were investigated against human fibrosarcoma HT-29 and human hepatocellular carcinoma HepG2. The treatment of 0.5 mg/ml of ethanol, ethanol after water and water extracts of P. linteus had the highest cytotoxicity of 59, 57, 54%, respectively against HT-27 cell line, whereas low cytotoxicity effects were observed against HepG2 cell line in the range of $10{\sim}30%$. The ethanol and water extracts of P. linteus also showed the nitrate scavenging ability at different pHs. The ethanol extract showed higher nitrate-scavenging ability compared to water extract of P. linteus.

• Journal of Life Science
• /
• v.21 no.11
• /
• pp.1518-1525
• /
• 2011

Sorafenib is the only approved systemic, therapeutic agent for hepatocellular carcinoma (HCC). The use of Ginseng Extract (GE) in cancer patients is growing worldwide; however, drug interaction between sorafenib and GE has not been illuminated. Four different human cancer cell lines including HepG2 were used and immunocompetent mice were implanted subcutaneously with a mouse HCC cell line. Treatment with low dose GE stimulated cell growth, while a high dose inhibited growth. pERK (phosphorylation of extracellular signal-regulated kinase) was concomitantly increased and decreased respective of different doses of GE. Antitumoral effect of sorafenib decreased in non-proliferating phase cells but was sensitized after low dose GE (LDG) treatment. PD98059 (ERK phosphorylation inhibitor) efficiently blocked ERK phosphorylation, resulting in loss of sorafenib sensitization even after LDG treatment. In the HCC mouse model, LDG alone slightly increased tumor size while sorafenib alone significantly decreased it. However, a combination of LDG and sorafenib significantly decreased tumor size compared with sorafenib alone. Increase of pERK was observed in some normal mice organs and mild inflammatory change was observed in some of these organs, suggesting pERK activation by LDG may cause unexpected toxicity in normal cells. GE, dose-dependently, induced stimulation or inhibition in some human cancer cell lines. Combinational use of GE and sorafenib possibly potentiated an antitumoral response to sorafenib. pERK level has been provided as a potential predictive marker for sorafenib. Our result may suggest GE's dual effects in relation to pERK level in HCC cancer cell lines, and that certain doses of GE can sensitize sorafenib.

• Nuclear Medicine and Molecular Imaging
• /
• v.41 no.3
• /
• pp.226-233
• /
• 2007

Purpose: Dual reporter gene imaging has several advantages for more sophisticated molecular imaging studies such as gene therapy monitoring. Herein, we have constructed hepatoma cell line expressing dual reporter genes of sodium iodide symporter (NIS) and enhanced green fluorescence protein (EGFP), and the functionalities of the genes were evaluated in vivo by nuclear and optical imaging. Materials and Methods: A pRetro-PN vector was constructed after separating NIS gene from pcDNA-NIS. RSV-EGFP-WPRE fragment separated from pLNRGW was cloned into pRetro-PN vector. The final vector expressing dual reporter genes was named pRetro-PNRGW. A human hepatoma (HepG2) cells were transfected by the retrovirus containing NIS and EGFP gene (HepG2-NE). Expression of NIS gene was confirmed by RT-PCR, radioiodine uptake and efflux studies. Expression of EGFP was confirmed by RT-PCR and fluorescence microscope. The HepG2 and HepG2-NE cells were implanted in shoulder and hindlimb of nude mice, then fluorescence image, gamma camera image and I-124 microPET image were undertaken. Results: The HepG2-NE cell was successfully constructed. RT-PCR showed NIS and EGFP mRNA expression. About 50% of cells showed fluorescence. The iodine uptake of NIS-expressed cells was about 9 times higher than control. In efflux study, $T_{1/2}$ of HepG2-NE cells was 9 min. HepG2-NE xenograft showed high signal-to-background fluorescent spots and higher iodine-uptake compared to those of HepG2 xenograft. Conclusion: A hepatoma cell line expressing NIS and EGFP dual reporter genes was successfully constructed and could be used as a potential either by therapeutic gene or imaging reporter gene.

• Journal of Life Science
• /
• v.25 no.3
• /
• pp.307-316
• /
• 2015

Beetle larvae have been used as a traditional medicine to treat various human liver diseases. To prove the liver protective function of Allomyrina dichotoma larvae (ADL), we induced liver damage by the intraperitoneal injection of a hepatotoxic reagent, diethylnitrosamine (DEN), to C3H/HeN male mice and orally administered freeze-dried ADL powder. ADL powder lessened DEN-induced hepatotoxicity considering the reduced signs of acute and chronic hepatotoxicities, such as the ALP level in the blood serum, TUNEL-positive hepatocytes, ductural reactions, steatotic hepatocytes, and collagen deposition of the Masson’s trichrome staining. In addition to hepatoprotection, the anti-cancer activity of ADL has been examined. The ADL powder was extracted with ethanol and then fractionated with hexane, ethyl acetate, and water by a solvent partition technique. The ethyl acetate fraction showed cytotoxicity to various cancer cells through induction of apoptosis and necrosis, as well as the perturbed metabolism of the cancer cell to trigger autophagy. Collectively, ADL contains bioactive substances that can protect hepatocytes from toxic chemicals and trigger cell death in cancer cells. Thus, further purification and analyses of ADL fractions could lead to the identification of novel bioactive compounds.

• Journal of Life Science
• /
• v.33 no.12
• /
• pp.1002-1014
• /
• 2023

The root of Cirsium japonicum var. maackii (Maxim.) has long been used in traditional medicine to prevent the onset and progression of various diseases and has been reported to exert a wide range of physiological effects, including antioxidant activity. However, research on its effects on hepatocytes remains scarce. This study used the human hepatocellular carcinoma HepG2 cell line to investigate the antioxidant activity of ethanol extract of C. japonicum root (EECJ) on hepatocytes. Hydrogen peroxide (H₂O₂) was used to mimic oxidative stress. The results showed that EECJ significantly reverted the decrease in cell viability and suppressed the release of lactate dehydrogenase in HepG2 cells treated with H₂O₂. Moreover, an analysis of changes in cell morphology, flow cytometry, and microtubule-associated protein light chain 3 (LC3) expression showed that EECJ significantly inhibited HepG2 cell autophagy induced by H₂O₂. Furthermore, it attenuated H₂O₂-induced apoptosis and cell cycle disruption by blocking intracellular reactive oxygen species and mitochondrial superoxide production, indicating strong antioxidant activity. EECJ also restored the decreased levels of intracellular glutathione (GSH) and enhanced the expression and activity of superoxide dismutase and GSH peroxidase in H₂O₂-treated HepG2 cells. Although an analysis of the components contained in EECJ and in vivo validation using animal models are needed, these findings indicate that EECJ is a promising candidate for the prevention and treatment of oxidative stress-induced liver cell damage.

• Korean Journal of Radiology
• /
• v.2 no.1
• /
• pp.28-36
• /
• 2001

Objective: To provide a systematic overview of the effects of various parameters on contrast enhancement within the same population, an animal experiment as well as a computer-aided simulation study was performed. Materials and Methods: In an animal experiment, single-level dynamic CT through the liver was performed at 5-second intervals just after the injection of contrast medium for 3 minutes. Combinations of three different amounts (1, 2, 3 mL/kg), concentrations (150, 200, 300 mgI/mL), and injection rates (0.5, 1, 2 mL/sec) were used. The CT number of the aorta (A), portal vein (P) and liver (L) was measured in each image, and time-attenuation curves for A, P and L were thus obtained. The degree of maximum enhancement (Imax) and time to reach peak enhancement (Tmax) of A, P and L were determined, and times to equilibrium (Teq) were analyzed. In the computed-aided simulation model, a program based on the amount, flow, and diffusion coefficient of body fluid in various compartments of the human body was designed. The input variables were the concentrations, volumes and injection rates of the contrast media used. The program generated the time-attenuation curves of A, P and L, as well as liver-to-hepatocellular carcinoma (HCC) contrast curves. On each curve, we calculated and plotted the optimal temporal window (time period above the lower threshold, which in this experiment was 10 Hounsfield units), the total area under the curve above the lower threshold, and the area within the optimal range. Results: A. Animal Experiment: At a given concentration and injection rate, an increased volume of contrast medium led to increases in Imax A, P and L. In addition, Tmax A, P, L and Teq were prolonged in parallel with increases in injection time The time-attenuation curve shifted upward and to the right. For a given volume and injection rate, an increased concentration of contrast medium increased the degree of aortic, portal and hepatic enhancement, though Tmax A, P and L remained the same. The time-attenuation curve shifted upward. For a given volume and concentration of contrast medium, changes in the injection rate had a prominent effect on aortic enhancement, and that of the portal vein and hepatic parenchyma also showed some increase, though the effect was less prominent. A increased in the rate of contrast injection led to shifting of the time enhancement curve to the left and upward. B. Computer Simulation: At a faster injection rate, there was minimal change in the degree of hepatic attenuation, though the duration of the optimal temporal window decreased. The area between 10 and 30 HU was greatest when contrast media was delivered at a rate of 2 3 mL/sec. Although the total area under the curve increased in proportion to the injection rate, most of this increase was above the upper threshould and thus the temporal window was narrow and the optimal area decreased. Conclusion: Increases in volume, concentration and injection rate all resulted in improved arterial enhancement. If cost was disregarded, increasing the injection volume was the most reliable way of obtaining good quality enhancement. The optimal way of delivering a given amount of contrast medium can be calculated using a computer-based mathematical model.