• Asian Pacific Journal of Cancer Prevention
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• v.14 no.3
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• pp.1787-1790
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• 2013

This study evaluated the effects of ZD1839, an orally active, selective epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, on nasopharyngeal carcinoma (NPC) both in vitro and in vivo. Influence of ZD1839 alone or combined with cisplatin on the NPC cell line CNE2 was detected by MTT assay with flow cytometry assessment of cell cycle distribution and apoptosis rates. Nude mice NPC xenografts were also used to evaluate the effects of ZD1839 alone or combined with cisplatin. The Student's t test evaluated statistical significance. ZD1839 alone or combined with cisplatin inhibited CNE2 cell line proliferation. ZD1839 induced CNE2 cell cycle arrest in the G1 phase, and higher concentrations induced apoptosis. Xenograft tumors were significantly smaller when treated with 200 mg/kg ZD1839, cisplatin, or cisplatin combined with 100 mg/kg ZD1839 than untreated controls. ZD1839 (200 mg/kg) alone showed good tumor inhibition effects, reduction of tumor weights, and smaller tumor volume without loss of body weight. ZD1839 (200 mg/kg) might provide a good and effective therapeutic reagent for NPC.

• Journal of Acupuncture Research
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• v.18 no.1
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• pp.88-99
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• 2001

Objectives : This study was performed to determine if Orostachys japonicus A. Berger aquacupuncture (OjB) provides the protective effect against the loss of celi viability and DNA damage induced by oxidant in renal proximal tubular cells. Methods : The cell viability was evaluated by a MTT reduction assay and DNA damage was estimated by measuring double stranded DNA breaks in opossum kidney (OK) cells, an established proximal tubular cell line. Lipid peroxidation was determined by measuring malondialdehyde (MDA), a product of lipid peroxidation. Results : $H_2O_2$ increased the loss of cell viability in a time-dependent manner, which were prevented by 0.1% OjB. The protective effect of OjB was dose-dependent over concentration range of 0.05-0.5%. $H_2O_2$ caused ATP depletion and DNA damage, which were prevented by OjB and the hydrogen peroxide scavenger catalase. The loss of cell viability by $H_2O_2$ was not affected by the antioxidant DPPD, but lipid peroxidation by the oxidant was completely inhibited by DPPD. Conclusions : These data suggest that $H_2O_2$-induced death results from a lipid peroxidation-independent mechanism and the protective effect of OjB is not associated with its antioxidant activity.

• Herbal Formula Science
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• v.17 no.1
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• pp.175-185
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• 2009

Petiolus Nelumbinis, branches of lotus leaf or lotus flower is a traditional oriental herbal medicine widely used for treating a superheat or disorder of qi flow. Although there are many clinical results and literature study, it has been rarely conducted to evaluate the immuno-biological activity. The present study was conducted to examine the anti-inflammatory effects of PNM (Petiolus Nelumbinis MeOH extract) in vitro. To determine cytotoxic concentration of PNM, the cells were treated with PNM for 24 h after LPS addition, and the cell viability was tested by MTT assay. Both of dosages (30 and 100 ${\mu}g/ml$) of PNM had no cytotoxicity. In these concentrations, PNM significantly reduced the elevated levels of NO and $PGE_2$ by LPS. These inhibitory effects of PNM were due to the reduced expressions of iNOS and COX-2 protein. TNF-$\alpha$, IL-1$\beta$ and IL-6 are frequently encountered pro-inflammatory cytokines, and LPS plays a key role in inducing to the massive production of these cytokines. Thus, we next determined the levels of these cytokines. Although PNM had no significant inhibitory effect on the production of TNF-$\alpha$, the elevated levels of IL-1$\beta$ and IL-6 by LPS were dose-dependently reduced in PNM-treated groups. These results demonstrate that PNM has anti-inflammatory effects by inhibiting the production of proinflammatory cytokines, NO and $PGE_2$ in LPS-activated macrophage. Moreover, the reduction of NO and $PGE_2$ levels was due to the inhibition of iNOS and COX-2 protein expression by PNM.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.10
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• pp.5849-5854
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• 2013

Background: Apoptosis may be induced after Bcl-2 expression is inhibited in proliferative cancer cells. This study focused on the effect of autophagy activation by ABT737 on anti-tumor effects of epirubicin. Methods: Cytotoxic effects of ABT737 on the HepG2 liver cancer cell line were assessed by MTT assay and cell apoptosis through flow cytometry. Mitochondrial membrane potential was measured by fluorescence microscopy. Monodansylcadaverin (MDC) staining was used to detect activation of autophagy. Expression of p53, p62, LC3, and Beclin1, apoptotic or autophagy related proteins, was detected by Western blotting. Results: ABT737 and epirubicin induced growth inhibition in HepG2 cells in a dose- and time-dependent manner. Both ABT737 and epirubicin alone could induce cell apoptosis with a reduction in mitochondrial membrane potential as well as increased apoptotic protein expression. Further increase of apoptosis was detected when HepG2 cells were co-treated with ABT373 and epirubicin. Furthermore, our results demonstrated that ABT373 or epirubicin ccould activate cell autophagy with elevated autophagosome formation, increased expression of autophagy related proteins and LC3 fluorescent puncta. Conclusions: ABT737 influences cancer cells through both apoptotic and autophagic mechanisms, and ABT737 may enhance the effects of epirubicin on HepG2 cells by activating autophagy and inducing apoptosis.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.10
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• pp.4357-4361
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• 2015

Thunbergia Laurifolia Linn. (TL) is one of the most familiar plants in Thai traditional medicine that is used to treat various conditions, including cancer. However, the antitumor activity of TL or its constituents has never been reported at the molecular level to support the folklore claim. The present study was designed to investigate the antitumor effect of an aqueous extract of TL in human breast cancer cells and the possible mechanism(s) of action. An aqueous crude extract was prepared from dried leaves of TL. Folin-Ciocalteu colorimetric assays were used to determine the total phenolic content. Antiproliferative and cell cycle effects were evaluated in human breast adenocarcinoma MCF-7 cells by MTT reduction assay, cell growth inhibition, clonogenic cell survival, and flow cytometric analysis. Free radical generation by the extracts was detected using electron paramagnetic resonance spectroscopy. The exposure of human breast adenocarcinoma MCF-7 cells to a TL aqueous extract resulted in decreases in cell growth, clonogenic cell survival, and cell viability in a concentration-dependent manner with an $IC_{50}$ value of $843{\mu}g/ml$. Treatments with extract for 24h at $250{\mu}g/ml$ or higher induced cell cycle arrest as indicated by a significant increase of cell population in the G1 phase and a significant decrease in the S phase of the cell cycle. The capability of the aqueous extract to generate radical intermediates was observed at both high pH and near-neutral pH conditions. The findings suggest the antitumor bioactivities of TL against selected breast cancer cells may be due to induction of a G1 cell cycle arrest. Cytotoxicity and cell cycle perturbation that are associated with a high concentration of the extract could be in part explained by the total phenolic contents in the extract and the capacity to generate radical intermediates to modulate cellular proliferative signals.

• Herbal Formula Science
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• v.28 no.1
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• pp.15-27
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• 2020

Objectives : We selected three herb-combined remedies, Bigihwan (BGH), Daechilgitang (DCGT) and Mokwhyangbinranghwan (MHBRH), through Donguibogam text-mining analysis, and evaluated their anti-cancer effects on human hepatocellular carcinoma Hep3B cells. Methods : Cytotoxicity was assessed by an MTT assay. Apoptosis rate, autophagy and ROS level were detected by flow cytometry. The autophagy was also observed by Cyto-ID staining fluorescence microscopy. The expression of autophagy, mitophagy and pexophagy regulatory proteins was detected by Western blot analysis. Results : BGH showed the strongest effect among the three prescriptions in inhibiting Hep3B cell viability, which was associated with the induction of apoptosis and autophagy. Autophagy blockers improved cell viability and reduced apoptosis after BGH and DCGT treatments, indicating that autophagy by these prescriptions enhanced Hep3B cells against their cytotoxicity. However, MHBRH enhanced the reduction of cell viability and apoptosis by autophagy blockers. Induction of autophagy by BGH treatment was associated with mitophagy due to mitochondrial dysfunction than DCGT and MHBRH-treated cells. In addition, induction of apoptosis by BGH treatment was ROS-dependent and showed the possibility of pexophagy involvement. Conclusion : Although further studies need to be conducted to study the efficacy and mechanism of accurate anticancer activity, the present results will serve as important sources of understanding the mechanism of action of herbal remedies prescribed for liver disease as documented in Donguibogam.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.2
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• pp.747-752
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• 2013

Oncostatin M (OSM) is a multifunctional cellular regulator acting on a wide variety of cells, which has potential roles in the regulation of gene activation, cell survival, proliferation and differentiation. Previous studies have shown that OSM can induce morphological and/or functional differentiation and maturation of many tumor cells. However, the action of OSM on the induction of differentiation of human hepatocellular carcinoma (HCC) has not been reported. Here, we investigated the effects of different concentrations of OSM on human HCC cell line SMMC-7721 growth, proliferation, cell cycling, apoptosis and differentiation in vitro. Cell growth was determined via MTT assay, proliferation by cell cycle analysis, apoptosis by flow cytometry, morphology by transmission electronic microscopy, and cell function by detection of biochemical markers. Our results demonstrated that OSM strongly inhibited the growth of SMMC-7721 cells in a dose-dependent manner, associated with decreased clonogenicity. Cell cycle analysis revealed a decreased proportion of cells in S phase, with arrest at G0/G1. The apotosis rate was increased after OSM treatment compared to the control. These changes were associated with striking changes in cellular morphology, toward a more mature hepatic phenotype, accompanied by significant reduction of the expression of AFP and specific activity of ${\gamma}$-GT, with remarkable increase in secretion of albumin and ALP activity. Taken together, our findings indicate that OSM could induce the differentiation and reduce cell viability of SMMC-7721 cells, suggesting that differentiation therapy with OSM offers the opportunity for therapeutic intervention in HCC.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.3
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• pp.599-607
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• 2009

Prunellae Spica is the spike or whole plant of Prunella vulgaris Linne, which has been used for clearing heat from the liver, brightening the eyes and treating headache in traditional oriental medicines. This study was conducted to evaluate the inhibitory effects of the aqueous extract of Prunellae Spica (PSE; PS extract) on the production of NO and PGE2 in LPS-activated Raw 264.7 cells. Cell viability was determined by MTT assay, and all three doses of PS extract (0.03, 0.1 and 0.3 mg/ml) had no significant cytotoxicity during the entire experimental period. The cells were treated with 1 ${\mu}g/ml$ of LPS 1 h before adding PS extract, and increased NO and PGE2 production were detected in LPS-activated cells compared to control. However, these increases were dose-dependently attenuated by treatment with PS extract. The inhibition of NO by PS extract was due to the suppression of iNOS expression via inhibition of $NF{\kappa}B$ nuclear translocation and proteolytic degradation of $I{\kappa}B{\alpha}$. The decreased level of PGE2 was derived from inhibition of COX-2 activity, but expression of COX-2 protein was not affected by PS extract. Moreover, PS extract reduced the elevated production of IL-${\beta}$ and IL-6 by LPS. These results demonstrate that PS extract has inhibitory effects on the production of NO and PGE2 as a consequence of the reduction of proinflammatory cytokines, especially IL-${\beta}$ and IL-6 in LPS-activated Raw 264.7 cells.

• Korean Journal of Medicinal Crop Science
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• v.22 no.5
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• pp.369-377
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• 2014

In order to find out anticancer activity of Korean pepper (Capsicum annuum L.), the cytotoxicity against 8 cell lines including 293 (normal kidney cells) and A-431 (epidermoid carcinoma cells) of extracts by extraction solvents and plant parts were investigated using MTT assay. Also the correlation between content of capsaicin known as anticancer ingredient and cytotoxicity of extracts from pepper were analyzed. The distilled water extracts from seed and germinated seed showed very high cytotoxicity against 6 cancer cell lines including A549 (lung carcinoma cells), AGS (stomach adenocarcinoma cells), HeLa (cervix adenocarcinoma cells), HepG2 (hepatoblastoma cells), HT-29 (colon adenocarcinoma cells), and MCF-7 (breast adenocarcinoma cells). But 80% ethanol and methanol extracts showed cytotoxicity against 293 and AGS. The $RC_{50}$, that was, the concentration of sample required for 50% reduction of cell viability, of seed and germinated seed extracts against AGS were $33.4{\sim}389.1{\mu}g/m{\ell}$ and $63.9{\sim}1,316.7{\mu}g/m{\ell}$, respectively, so anticancer activity was higher in seed than in germinated seed. In capsaicin contents, seed with high cytotoxicity and pericarp with a little cytotoxicity contained $47.4{\sim}1,260.0{\mu}g/g$ and $58.3{\sim}1,498.0{\mu}g/g$, respectively. As these results, the correlation was not between cytotoxicity and capsaicin content.

• Korean Journal of Food Science and Technology
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• v.43 no.3
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• pp.396-400
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• 2011

We examined the in vitro mutagenic and antimutagenic effects of hexane-extracted hemp (Cannabis sativa L. subsp. sativa var. sativa) seed oil (HO) with and without S9-mediated metabolic activation, using the TA98 and TA100 Salmonella Typhimurium strains. The MTT assay revealed no cytotoxicity in HepG2 cells for HO quantities $\leq400g$/mL. In the mutagenicity test, revertant colonies did not exceed spontaneous colonies in number. Colony numbers did not increase in either strain after HO treatment, with or without metabolic activation. HO showed no mutagenic effects and did not induce a mutation in either strain. In the antimutagenicity test, HO reduced the number of mutated colonies induced by 4NQO in both strains. The inhibition rates of HO (TA98, 21-91%; TA100, 21-85%) indicated a potent reduction in mutagenicity induced by 4NQO. HO showed no significant mutagenicity and may have antimutagenic effects, as assessed by Ames testing.