• Microbiology and Biotechnology Letters
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• v.17 no.3
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• pp.231-235
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• 1989

Possibility of using microcarriers for the growth of a transformed human embryonic kidney cell line 293 was investigated. The cell grew well in a static culture such as T-flasks with medium of DME/F12 (3:1) mixture supplemented with 5% FBS, but it was most difficult to make the cells grow on microcarriers mainly due to the low attachment efficiency and poor spreading at initial stage of the culture. Consequently, 30-50% of the cells were lost upon inoculation into microcarrier suspension and significant fraction of the mirrocarrier became bald. The medium supplemented with the concentrated conditioned medium by hepatoma cell line HpG2 supported the active growth of the cells on microcarrier and the cells showed a very healthy and well spreading morphology. It was probable that some spreading and attachment factors of HpG2 conditioned medium were effective for 293 cells.

• Journal of Life Science
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• v.15 no.3 s.70
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• pp.323-331
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• 2005

The objective of the present study was to investigate the effect of $\beta-lapachone$, a quinone obtained from the bark of the lapacho tree (Tabebuia avellanedae) in South America, on the cell growth of human hepatoma (HepG2) and bladder (T24) carcinoma cells. Exposure of cancer cells to $\beta-lapachone$ resulted in growth inhibition, morphological changes and apoptosis in a concentration-dependent manner, which could be proved by MTT assay and flow cytometry analysis. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analyses revealed that $\beta-lapachone$ did not affect the levels of tumor suppressor p53 and cyclin-dependent kinase (Cdk) inhibitor p21 (WAFl/CIPl) expression. However, the transcriptional factor Sp-l and proliferating cell nuclear antigen (PCNA) protein levels were significantly down-regulated by $\beta-lapachone$ in both cell lines. Moreover, $\beta-lapachone$ treatment caused a dose-dependent inhibition of the expression of telomere regulatory gene products such as human telomere reverse transcriptase (hTERT) and telomerase-associated protein-l (TEP-l). Taken together, these findings suggest that $\beta-lapachone$-induced inhibition of human hepatoma and bladder carcinoma cell proliferation is associated with the induction of apoptotic cell death via modulation of several major growth regulatory gene products, and provide important new insights into the additional mechanisms of the anti-cancer activity of $\beta-lapachone$.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.167.1-167.1
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• 2003

Here we show that panaxadiol, a ginseng saponin with a dammarane skeleton, induces acute apoptotic cell death in human hepatoma SK-HEP-1 cells as evidenced by analysis of DNA fragmentation, caspase activation, and changes in cell morphology. The kinetic study showed that panaxadiol-induced apoptosis is associated with depolarization of mitochondrial membrane potential and cytochrome c release. Sequential activations of caspases-depolarization of mitochondrial membrane potential and cytochrome c release. Sequential activations of caspases-9, and -3, or -7, but not of caspase 8 coincide well in a time dependent manner with mitochondrial membrane depolarization and cytochrome c release from mitochondria during apoptosis of SK-HEP-1 cells induced by treatment with panaxadiol. (omitted)

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.3
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• pp.1423-1428
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• 2014

Iso-suillin, a natural product isolated from Suillus luteus, has been shown to inhibit the growth of some cancer cell lines. However, the molecular mechanisms of action of this compound are poorly understood. The purpose of this study was to investigate how iso-suillin inhibits proliferation and induces apoptosis in a human hepatoma cell line (SMMC-7721). We demonstrated the effects of iso-suillin on cell proliferation and apoptosis in SMMC-7721 cells, with no apparent toxicity in normal human lymphocytes, using colony formation assays and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) analysis. Western blotting was used to examine the expression of $G_1$ phase-regulated and apoptosis-associated protein levels in iso-suillin treated SMMC-7721 cells. The results indicated that iso-suillin significantly decreased viability, induced $G_1$ phase arrest and triggered apoptosis in SMMC-7721cells. Taken together, these results suggest the potential of iso-suillin as a candidate for liver cancer treatment.

• Archives of Pharmacal Research
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• v.27 no.12
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• pp.1245-1252
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• 2004

Although ascorbate (vitamin C) has been shown to have anti-cancer actions, its effect on human hepatoma cells has not yet been investigated, and thus, the exact mechanism of this action is not fully understood. In this study, the mechanism by which ascorbate induces apoptosis using HepG2 human hepatoblastoma cells is investigated. Ascorbate induced apoptotic cell death in a dose-dependent manner in the cells, was assessed through flow cytometric analysis. Contrary to expectation, ascorbate did not alter the cellular redox status, and treatment with antioxidants (N-acetyl cysteine and N,N-diphenyl-p-phenylenediamine) had no influence on the ascorbate-induced apoptosis. However, ascorbate induced a rapid and sustained increase in intracellular $Ca^{2+}$ concentration. EGTA, an extracellular $Ca^{2+}$ chelator did not significantly alter the ascorbate-induced intracellular $Ca^{2+}$ increase and apoptosis, whereas dantrolene, an intracellular $Ca^{2+}$ release blocker, completely blocked these actions of ascorbate. In addition, phospholipase C (PLC) inhibitors (U-73122 and manoalide) significantly suppressed the intracellular $Ca^{2+}$ release and apoptosis induced by ascorbate. Collectively, these results suggest that ascorbate induced apoptosis without changes in the cellular redox status in HepG2 cells, and that the PLC-coupled intracellular $Ca^{2+}$ release mechanism may mediate ascorbate-induced apoptosis.

• Food Science and Biotechnology
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• v.14 no.2
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• pp.297-300
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• 2005

Effects of allyl sulfides on kinetic behavior of cytochrome P450 1 (CYP1)-catalyzed ethoxyresorufin O-deethylase (EROD) activity were studied using microsomes from benzo[a]pyrene-treated human hepatoma cells. Apparent $K_m$ and $V_{max}$ values were calculated as $2.8\;{\mu}M$ and $3.0\;{\mu}mol$ resorufin/min/mg protein based on Lineweaver-Burk plot of microsomal EROD activity, respectively. Diallyl disulfide (DADS) and diallyl trisulfide (DATS) affected $K_m$ and $V_{max}$ values of EROD activity and acted as mixed-type inhibitors for CYP1 isozymes. Apparent Ki values of DADS and DATS were calculated as 1.07 and 0.88 mM, respectively, by re-plotting slopes of Lineweaver-Burk plot and inhibitor concentrations.

• Archives of Pharmacal Research
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• v.28 no.1
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• pp.73-80
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• 2005

The effect of capsaicin on apoptotic cell death was investigated in HepG2 human hepatoma cells. Capsaicin induced apoptosis in time- and dose-dependent manners. Capsaicin induced a rapid and sustained increase in intracellular $Ca^{2+}$ concentration, and BAPTA, an intracellular $Ca^{2+}$ chelator, significantly inhibited capsaicin-induced apoptosis. The capsaicin-induced increase in the intracellular $Ca^{2+}$ and apoptosis were not significantly affected by the extracellular $Ca^{2+}$ chelation with EGTA, whereas blockers of intracellular $Ca^{2+}$ release (dantrolene) and phospholipase C inhibitors, U-73122 and manoalide, profoundly reduced the capsaicin effects. Interestingly, treatment with the vanilloid receptor antagonist, capsazepine, did not inhibit either the increased capsaicin-induced $Ca^{2+}$ or apoptosis. Collectively, these results suggest that the capsaicin-induced apoptosis in the HepG2 cells may result from the activation of a PLC-dependent intracellular $Ca^{2+}$ release pathway, and it is further suggested that capsaicin may be valuable for the therapeutic intervention of human hepatomas.

• Journal of Microbiology and Biotechnology
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• v.14 no.5
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• pp.891-900
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• 2004

UDP-N-Acetylglucosamine(GlcNAc):$\beta$1,4-D-mannoside$\beta$-l ,4N-acetylglucosaminyltransferase-III (GnT-III) and UDP-N-GlcNAc:$\alpha$-6-D-mannosid$\beta$-1,6N-acetylglucosaminyltransferase-V(GnT - V) activities were determined in human hepatoma cell lines and metastatic colon cancer cells, and their activities were compared with those of normal liver cells and fetal hepatocytes. GnT-III activities were higher than those of GnT-V in hepatic carcinoma cells. When the two enzyme activities were assayed in highly metastatic colon cancer cells, GnT - V activities were much higher than those of GnT-III. When GlcN, GlcN-biant-PA and UDP-GlcNAc were used as substrates, the enzymes displayed different kinetic properties between hepatic and colon cancer cells, depending on their metastatic potentials. Normal cells of two origins had characteristically very low levels of GnT-III and -V activities, whereas hepatoma and colon cancer cells contained high levels of activities. These data were supported by RT-PCR and Northern blot analyses, showing that the expression of GnT-III and -V mRNAs were increased in proportion to the enzymatic activities. The increased GnT-III, md -V activities were also correlated with increased glycosylation of the cellular glycoproteins in hepatoma and colon cancer cells, as examined by lectin blotting analysis by using wheat germ glutinin (WGA), erythroagglutinating phytohemagglutinin (E-PHA), leukoagglutinating phytohemagglutinin (L-PHA), and concanavalin A (Con A). Treatment with retinoic acid, a differentiation agent, resulted in decreases of both GnT-III and -V activities of HepG2 and HepG3 cells. In colon carcinoma cells, however, treatment with retinoic acid resulted in a reduction of GnT-V activity, but not with GnT-III activity. Although the mechanism underlying the induction of these mzymes is unclear, oligosaccharides in many glycoproteins have been observed of cancer cells.

• The Journal of Korean Medicine
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• v.27 no.4
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• pp.48-56
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• 2006

Objectives : This study is aimed to elucidate anti-hepatoma activity of Coptis Chinensis Extract (CCE) and evaluate its effect on proliferation of human hepatoma Hep G2 cells. Methods : To identify CCE and control the quality, we performed fingerprinting by high-performance thin layer chromatography (HPTLC). To investigate effects of CCE on anti-hepatoma activity, we measured cytotoxicity against Hep G2 cells compared with treatment of paclitaxel and 5-fluorouracil (5-FU). To examine the mechanism of inhibitory effect of CCE on Hep G2 cell proliferation, cell cycle distribution was evaluated using fluorescent activated cell sorter (FACS) Result : CCE showed a significant effect that arrests Hep G2 cells at the G2/M phase of the cell cycle. CCE combined with paclitaxel inhibited synergistically cell growth of Hep G2 cells. Conclusion : CCE may present anticancer effects through inhibition of hepatocellular carcinoma (HCC) cell proliferation via G2/M arrest, and may be a useful anticancer agent for HCC.

• Korean Journal of Plant Resources
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• v.25 no.5
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• pp.647-651
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• 2012

The objective of this study was to investigate the anticancer effects of the extracts of oriental melon seeds. Various solvent extracts of oriental melon seeds were prepared and their anticancer effects were examined using in vitro MTT and CV assays. The anticancer effects of various extracts of oriental melon seeds were also examined in five human cancer cell lines including A549, AGS, HT-29, MCF-7 and HepG2. The ethanol extract of heated oriental melon seeds showed the potent cytotoxic effects especially against mouse hepatoma cell line(Hepa1c1c7), human hepatoma cell line(HepG2) and human breast cancer cell line(MCF-7). These data suggest that oriental melon seeds can be a promising anticancer agent against human liver and breast cancer.