• Asian Pacific Journal of Cancer Prevention
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• v.17 no.sup3
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• pp.125-130
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• 2016

Nepeta cataria L. has been used in traditional medicine of some countries. Here the cytotoxic and apoptogenic activity of methanol extracts, n-hexane, dichloromethane, ethyl acetate, n-butanol, and acqueous extracts and the essential oil obtained from the aerial parts of the plant were evaluated with PC3, DU-145 and MCF-7 cell lines. Cell viability, histograms of PI stained fragmented DNA in apoptotic cells and Western blot analysis of proteins involved in the cascade of apoptosis were compared in all samples. Thirty components were identified as volatile, representing 99.7% of essential oil composition after GC-MS analysis of the oil obtained from aerial parts of the N. cataria by hydro-distillation. The major oil components of the essential oil were nepetalactone stereoisomers. Comparing IC50 values showed estrogen receptor positive PC3 cells were more sensitive to the cytotoxic effects of N. cataria in comparison with low hormone-receptor presenting DU-145 cells. Among multiple extracts and essential oils of the plant, only the ethyl acetate extract could significantly decrease cell viability in PC3 cells, in a concentration dependent manner. Ethyl acetate extract of N. cataria treated cells showed a sub-G1 peak in PC3 cells in a concentration dependent manner that indicates the involvement of an apoptotic process in ethyl acetate extract-induced cell death. Western blotting analysis showed that in PC3 cells treated with ethyl acetate (48 h) caspase 3 and PARP were cleaved to active forms. Overall, the results suggest that further analytical elucidation of N. cataria in respect to finding new cytotoxic chemicals with anti-tumor activity is warranted.

• Korean Journal of Pharmacognosy
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• v.40 no.1
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• pp.6-12
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• 2009

Thirty five methanol extracts from 19 natural local plants, which have been used as traditional anti-cancer medicine, were prepared. They were analyzed the cytotoxic effects on primary fibroblast cells and carcinoma cells. The root extract of Solanum nigrum were highly toxic in both cell lines with $IC_{50}$ values of less than $0.01{\mu}g/{\mu}l$, and 26 of 35 extracts were toxic in all cells with $IC_{50}$ values of $0.1{\sim}2{\mu}g/{\mu}l$. Three extracts including the fruit extracts of Solanum nigrum and Morus alba had no cytotoxic activity in both cell lines. Five of 35 extracts were highly toxic in cancer cells than in primary cells. Because primary cells were more resistant on these extracts, the five extracts were selected for anti-cancer agent candidates. Apoptosis or programmed cell death has an essential role in chemotherapy-induced tumor cell killing. Recently, inducers of apoptosis have been used in cancer therapy. When two of 5 cancer cell-specific cytotoxic extracts (Ulmus parvifolia and Zelkova serrata) were treated in concentration of $0.02{\sim}0.1{\mu}g/{\mu}l$, apoptosis were increased at 3-5 times in cancer cell lines. Finally, the apoptotic effects of these extracts were confirmed by cleavages of both poly-(ADP-ribose)-polymerase and caspase-3 as apoptotic markers. In this report, we suggested that two of 35 medicinal herb extracts can be useful anti-cancer drug candidates inducing apoptosis in several carcinoma cell lines.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.2
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• pp.290-295
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• 2008

The cytotoxic characteristics of Vitsin A isolated from Vitis amurensis L. were examined in human colorectal, breast, uterine and renal cancer cells. Vitsin A showed good cytotoxicity against various cancer cells with $IC_{50}$ of $1\;{\sim}\;30\;{\mu}M$. Among them, Vitisin A exhibited strongest cytotoxic effect against MES-SA cells with $IC_{50}$ of 1.11 ${\mu}M$ by SRB assay. To verify whether the cytotoxicity of Vitisin A may be associated with apoptosis, TdT-mediated-dUTP Nick-End Labeling (TUNEL) assay and cell cycle analysis were performed in MES-SA cells. Apoptotic bodies were observed in Vitisin A treated MES-SA cells by TUNEL assay. Also, Vitisin A effectively increased the portion of $sub-G_1$ DNA content by flow cytometric analysis. Taken together, these findings suggest that the cytotoxicity of Vitisin A against MES-SA cells is chiefly mediated by apoptosis.

• The Journal of Korean Obstetrics and Gynecology
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• v.19 no.1
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• pp.97-110
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• 2006

Purpose : To address the ability of Myrrha (MY) to induce cell death, we investigated the effect of MY on apoptosis. In human cervical carcinoma HeLa cells, apoptosis occurred following MY exposure in a dose-dependent manner. Methods : We have tested several kinds of anti-oxidants to investigate the MY-induced apoptotic mechanism. Among the anti-oxidants, N-acetyl cysteine(NAC) or reduced glutathione (GSH) protects MY-induced apoptosis. NAC is an aminothiol and synthetic precursor of intracellular cysteine and GSH. To confirm the role of GSH in MY-induced apoptosis, methionine and cystathionine-glutathione extrusion inhibitors were treated in the presence of MY. Results : NAC, GSH, methionine or cystathionine led to protective effect against MY-induced apoptosis in HeLa cells. The GSH and GSH-associated reagents regulate MY-induced cytochrome c release and the resultant caspase-3 activation. Furthermore, the two specific inhibitors of carrier-mediated GSH extrusion, methionine and cystathionine demonstrate GSH extrusion occurs via a specific mechanism. While decreasing GSH extrusion and protecting against MY-induced apoptosis, methionine and cystathionine failed to exert anti-apoptotic activity in cells previously deprived of GSH. Conclusion : the target of the protection is indeed GSH extrusion. This shows that the protective effect is achieved by forcing GSH to stay within the cells during apoptogenic treatment. All this evidence indicates the extrusion of GSH precedes andis responsible for the apoptosis, probably by altering the intracellular redox state, thus giving a rationale for the development of redox-dependent apoptosis in MY-treated human cervical carcinoma HeLa cells.

• Animal cells and systems
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• v.2 no.3
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• pp.403-410
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• 1998

The Tcf-1 (T cell specific factor-1) is a transcription factor uniquely expressed in T-lineage cells. Its expression is developmentally regulated, which is high in the specific stage of immature thymocytes, but is much lower in mature T cells. We cloned the Tcf-1 gene by subtractive hybridization and found it to be highly expressed in the thymus compared to the mRNA level in the spleen as expected. Since apoptosis occurs enormously in the thymus, we were interested in whether Tcf-1 gene expression could be regulated by such a high level of apoptotic assault. By using T cell hybridoma 70.7 cells, we induced apoptosis by incubating cells with anti-CD3 antibody in vitro. After apoptosis induction, Tcf-1 mRNA level was found to be significantly reduced compared to normal cells. Since Tcf-1 is a transcription factor for the CD3-e gene, we tested how CD3-e expression is regulated in apoptotic cells. The surface level of CD3-e protein is also down-regulated after apoptosis induction. Such a down-modulation of CD3-e protein would reduce the TCR/CD3 complex on the cell surface, which would be an important regulator for T cell apoptosis.

• The Journal of Korean Medicine
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• v.28 no.1 s.69
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• pp.198-210
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• 2007

Objectives : Nitric oxide(NO) is a reactive free radical and a messenger molecule in many physiological functions. However, excessive release NO of induces neurotoxicity. We investigated whether a mixture of red ginseng and paeonia radix prossesses a protective effect against sodium nitroprusside(SNP)-induced apoptosis in the human neuroblastoma cell line SK-N-MC. Methods : We performed 3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay, 4,6-diamidino-2-phenylindole(DAPD) staining, terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick end labeling(TUNEL)assay, DNA fragmentation assay, reverse transcription-polymerase chain reaction(RT-PCR), Western blot analysis, and caspase-3 enzyme activity assay in SK-N-HFC cells. Result : MTT assay showed that SNP treatment significantly reduced the viabilities of cells and that pre-treatment with the red ginseng and paeonia radix mixture alleviated SNP-induced cytotoxicity. The cells treated with SNP exhibited several apoptotic features, while those pre-treated fir 1 h with the mixture of red ginseng and paeonia radix 1 h prior to SNP expose showed reduced apoptotic features. In addition, the cells pre-treated with the red ginseng and paeonia radix mixture for 1 h prior to SNP expose increased bel-2 expressions, decreased Bax expressions, and decreased caspase-3 enzyme activity. Conclusions : These results show that the red ginseng and paeonia radix mixture exerts a protective effect against SNP-induced apoptosis in SK-N-MC cells.

• Journal of Embryo Transfer
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• v.31 no.1
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• pp.89-95
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• 2016

The present study was conduct to examine the $H_2O_2$ expression level and apoptosis-related gene expression levels inporcineskin-derived stem cell-like cells (pSSCs) after adipogenic, chondrogenic, and osteogenic differentiation induction. The pSSCs were obtained by digestion of porcine ear skin biopsy and cultured in each induction medium for 21 to 26 days to induce adipogenic, chondrogenic, and osteogenic differentiation, respectively. The $H_2O_2$ levels of pSSCs after induction culture were evaluated by staining with 2'7'-dichlorodihydrofluorescein diacetate ($H_2DCFDA$). The apoptotic gene expression of pSSCs after induction culture was also estimated by RT-PCR. The pSSCs have a potential to differentiate into three mesodermal cell types (adipocytes, chondrocytes, and osteoblasts). Non-induced control and chondrogenic-induced cells were showed higher $H_2DCFDA$ intensity (P<0.05) than adipogenic- and osteogenic-induced cells. The relative expression of Bax/Bcl-2 level was significantly low (P<0.05) in adipogenic- and osteogenic-induced cells compared to non-induced control. However, there was no difference in the relative expression of Bax/Bcl-2 level among differentiation induction groups. The result of the present study shows that the apoptosis of pSSCs is not detrimentally increased by differentiation induction culture, although chondrogenic-induced pSSCs showed high ROS generation level and apoptotic index similarly to those of non-induced cells.

• Korean Journal of Pharmacognosy
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• v.44 no.3
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• pp.269-274
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• 2013

In this study, we investigated the anticancer effect of rice bran extract in HL-60 human promyelocytic leukemia cells. The extract of rice bran inhibited the proliferation of HL-60 cells. When treated with the rice bran extract, we could observe the apoptotic characteristics such as apoptotic bodies and the increase of sub-G1 hypodiploid cell population, increase of Bax level, decrease of Bcl-2 expression, cleavage of procaspase-3, cleavage of procaspase-9 and cleavage of poly(ADP-ribose) polymerase(PARP) in HL-60 cells. Furthermore, the apoptosis induction of HL-60 cells treated with the rice bran extract was also accompanied by the inactivation of mitogen-activated protein kinases (MAPK) such as ERK1/2 MAPK and p38 MAPK. In addition, the rice bran extract induced the down-regulation of c-myc. These data suggested that the rice bran extract could induce the apoptosis via the inactivation of ERK1/2 MAPK and p38 MAPK, and the down-regulation of c-myc in HL-60 acute pomyelocytic leukemia cells. The results support that the rice bran extract might have potential for the treatment of acute promyelocytic leukemia.

• Parasites, Hosts and Diseases
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• v.51 no.1
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• pp.61-68
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• 2013

Entamoeba histolytica, which causes amoebic colitis and occasionally liver abscess in humans, is able to induce host cell death. However, signaling mechanisms of colon cell death induced by E. histolytica are not fully elucidated. In this study, we investigated the signaling role of NOX in cell death of HT29 colonic epithelial cells induced by E. histolytica. Incubation of HT29 cells with amoebic trophozoites resulted in DNA fragmentation that is a hallmark of apoptotic cell death. In addition, E. histolytica generate intracellular reactive oxygen species (ROS) in a contact-dependent manner. Inhibition of intracellular ROS level with treatment with DPI, an inhibitor of NADPH oxidases (NOXs), decreased Entamoebainduced ROS generation and cell death in HT29 cells. However, pan-caspase inhibitor did not affect E. histolytica-induced HT29 cell death. In HT29 cells, catalytic subunit NOX1 and regulatory subunit Rac1 for NOX1 activation were highly expressed. We next investigated whether NADPH oxidase 1 (NOX1)-derived ROS is closely associated with HT29 cell death induced by E. histolytica. Suppression of Rac1 by siRNA significantly inhibited Entamoeba-induced cell death. Moreover, knockdown of NOX1 by siRNA, effectively inhibited E. histolytica-triggered DNA fragmentation in HT29 cells. These results suggest that NOX1-derived ROS is required for apoptotic cell death in HT29 colon epithelial cells induced by E. histolytica.

• The Korea Journal of Herbology
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• v.30 no.2
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• pp.25-30
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• 2015

Objectives : Laver (Porphyra tenera), a red algae species, is one of the most widely consumed edible seaweed in Korea. Laver contains various substances such as essential amino acid, fiber, minerals and polyphenols that benefit human health. In the present study, we prepared ethanol extracts from commercially processed product of Porphyra tenera, and evaluated the growth inhibitory effect against human oral squamous carcinoma YD-10B cells. Methods : Cell viability was measured by MTT assay. Apoptosis was confirmed by TUNEL assay and flow cytometry with the green fluorescent dye FITC annexin V entering apoptotic cells and the red fluorescent dye PI not entering. The expression of the relevant proteins was detected using Western blot. Results : Ethanol extracts of Porphyra tenera (PTE, $50-200{\mu}g/m{\ell}$) caused a significant decrease of cell viability in a dose dependant manner. The cell death occurred as a result of apoptotic process as determined by TUNEL assay and flow cytometric analysis. In line with this observation, decrease in procaspase proteins and increase in cytosolic cytochrome c were observed in cells treated with PTE. In addition, exposure to PTE decreased the expression levels of Bcl-2, and induced PARP cleavage and AIF translocation from mitochondria to nucleus. Conclusions : In conclusion, PTE exerts anti-cancer effects by inducing apoptosis via caspase activation and AIF nuclear translocation in YD-10B cells. These results provide evidence for the possible therapeutic effect of Porphyra tenera in oral cancer cells.