• The Korea Journal of Herbology
• /
• v.31 no.1
• /
• pp.77-82
• /
• 2016

Objectives : The study was designed to observe the effect of Polygalae Radix Preparata Cum Glycyrrhizae Radix on 4-Hydroxynonenal (4-HNE)-induced apoptosis in PC-12 cell.Methods : A cytotoxic test on Polygalae Radix Preparata Cum Glycyrrhizae Radix (PG) was conducted and another MTT assay was conducted to observe the cytoprotective effect against 4-HNE that cause oxidative stress. In addition, in order to observe the expression of Bax, Bcl-2, Caspase-3 and TNF-α protein involved with apoptosis, western blot was conducted.Results : The groups treated with 25 ㎍, 50 ㎍ and 100 ㎍ of PG water extract had no toxicity for PC-12 cell. The groups treated with 25 ㎍, 50 ㎍ and 100 ㎍ of PG water extract showed a significant increase of cell survival rate in comparison with the control group injected by only 4-HNE. The groups treated with 25 ㎍ and 50 ㎍ of PG water extract showed a significant supression on increase of Bax protein expression in the control group. The group treated with 100 ㎍ of PG water extract showed a significant promotion on decrease of Bcl-2 protein expression in the control group. The group treated with 50 ㎍ of PG water extract showed a significant supression on increase of Caspase-3 protein expression in the control group. The group treated with 25 ㎍ of PG water extract showed a significant supression on increase of TNF-α protein expression in the control group.Conclusions : These results suggest that Polygalae Radix Preparata Cum Glycyrrhizae Radix is effective in reducing apoptosis by 4-HNE-dameged cell.

• Asian Pacific Journal of Cancer Prevention
• /
• v.15 no.24
• /
• pp.10825-10830
• /
• 2015

The present study was conducted to investigate effects and mechanisms of action of phenolic alkaloids of Menispermum dauricum (PAMD) on gastric cancer in vivo. In vitro, cell apoptosis of human gastric cancer cell line SGC-7901 was observed using fluorescence staining. In vivo, a mice model was constructed to observe tumor growth with different doses. Cell apoptosis was examined using flow cytometry and K-RAS protein expression using Western blotting. The mRNA expression of P53, BCL-2, BAX, CASPASE-3, K-RAS was examined by real-time PCR. PAMD significantly suppressed tumor growth in the xenograft model of gastric cancer in a dose-dependent manner (p<0.01). Functionally, PAMD promoted cell apoptosis of the SGC-7901 cells and significantly increased the rate of cell apoptosis of gastric tumor cells (p<0.05). Mechanically, PAMD inhibited the expression of oncogenic K-RAS both at the mRNA and protein levels. In addition, PAMD affected the mRNA expression of the cell apoptosis-related genes (P53, BCL-2, BAX, CASPASE-3). PAMD could suppress gastric tumor growth in vivo, possibly through inhibiting oncogenic K-RAS, and induce cell apoptosis possibly by targeting the cell apoptosis-related genes of P53, BCL-2, BAX, CASPASE-3.

• The Journal of Internal Korean Medicine
• /
• v.27 no.2
• /
• pp.429-443
• /
• 2006

Objectives : The Purpose of this study was to identify anti-tumor effects of Curcuma longa L. on some kinds of cancer cells through molecular biologic methods. Materials & Methods : We used 4 kinds of cancer cell lines such as glioma cells(A172), cervical cancer cells(HeLa), Prostate cancer cells(PC3), lung cancer cells(A549). We injected the boiled extract of Curcuma longa L. $5{\mu}g,\;10{\mu}g$ to culture media(ml) for 24 hours. We measured the cytotoxic effect on 4 kinds of cancer cells through trypan blue exclusion test and the suppressive effect on viability of 4 kinds of cancer cells via MTT assay. We measured the change of mitochondria membrane potential via flow cytometry. The quantitative RT-PCR was used to examine the effect on the revelation of Bcl-2 and Bax which genes are related to apoptosis. We examined the effect on the revelation of Bcl-2 protein and Bax protein by western blot analysis. Results: 1. Extract of Curcuma longa L. showed significant cytotoxic effect on A172, HeLa, PC3 compared to the control group with density dependent manner. 2. Extract of Curcuma longs L. showed significant suppressive effect on viability of A172, HeLa, PC3 compared to the control group with density dependent manner. 3. Curcuma longs L. induced apoptosis by decreasing the membrane potential of mitochondria in A172, HeLa, PC3. 4. In the test about the revelation of genes related to apoptosis, the revelation of Bcl-2 decreased and the revelation of Bax increased in A172. HeLa, PC3 treated with Curcuma longa L. with density dependent manner. 5. In the test about the revelation of protein related to apoptosis, the protein levels of Bcl-2 decreased and the protein levels of Bax increased in A172, HeLa, PC3 treated with Curcuma longa L. Conclusions: This experiment shewed that Curcuma longs L. has anti-tumor effect on glioma, cervical, Prostate cancer cells except on lung cancer. We hope that anti-tumor effects of Curcuma longa L. will be more Practically identified.

• Journal of Chest Surgery
• /
• v.36 no.11
• /
• pp.852-857
• /
• 2003

Background: Conventional treatment for mesothelioma is largely ineffective. We evaluated the novel approach of adenoviral gene transfection of PTEN gene in mesothelioma cancer cell lines, inflammatory and epithelial subtype, which are sensitive to adenoviral p53. Material and Method: Binary adenoviral PTEN and LacZ (Ad/GT-LacZ and Ad/GV16) vectors were used for transduction of the mesothelioma cell lines, REN (p53 sensitive). Protein levels were determined by Western blotting assay. Apoptosis was assessed by fluorescence-activated cell sorter analysis of subdiploid populations. Cell viability was determined with the XTT assay. Statistical analysis was performed with analysis of variance and the Student t test. Result: 72 hours after the treatment of adenoviral PTEN gene, cell killing were 32.9% for REN compared to control cell (2.5%) at MOI of 20. Also we observed the over-expression of proapoptotic protein, bax and decreased expression of bcl-2 protein in REN cells. But the expression of BCL-xl, Bak, Bad proteins were not altered. Conclusion: Adenovirus Pten-mediated overexpression of the Bax gene induces apoptosis and decreased cellular viability in p53-sensitive mesothelioma cells. These data suggest that the transfection of PTEN gene may represent a alternative gene therapy strategy to treat mesothelioma.

• BMB Reports
• /
• v.35 no.5
• /
• pp.452-458
• /
• 2002

Exposure to ultraviolet light (UV) induces apoptosis in mammalian cells, The caspase group of proteases is required for the appotosis. This pathway is initiated by a release of cytochrome c from the mitochondria into the cytosol. Several Bcl-2 family proteins can regulate the release of cytochrome c by stabilizing the mitochondrial membrane. Here we show that expression of the endogenous bcl-xL was strongly downregulated in NIH3T3 cells within 2 h after UV-C irradiation, and that of bax was upregulated from 8 h after irradiation. Apoptosis was induced in more than 50% of the NIH3T3 cells 48 h after irradiation. Constitutive overexpression of bcl-xL in NIH3T3 cells protected the UV-induced apoptosis by preventing the loss of mitochondrial membrane potential and the activation of caspase 9. There results suggest that downregulation of Bcl-xL is relevant to UV-induced apoptosis of tibroblasts.

• BMB Reports
• /
• v.40 no.4
• /
• pp.475-485
• /
• 2007

Methamphetamine is an illicit drug that is often abused and can cause neuropsychiatric and neurotoxic damage. Repeated administration of psychostimulants such as methamphetamine induces a behavioral sensitization. According to a previous study, Bax was involved in neurotoxicity by methamphetamine, but the function of Bax in rewarding effect has not yet been elucidated. Therefore, we have studied the function of Bax in a rewarding effect model. In the present study, we treated chronic methamphetamine exposure in a Bax-deficient mouse model and examined behavioral change using a conditioned place preference (CPP) test. The CPP score in Bax knockout mice was decreased compared to that of wild-type mice. Therefore, we screened for Bax-related genes that are involved in rewarding effect using microarray technology. In order to confirm microarray data, we applied the RT-PCR method to observe relative changes of Bcl2, a pro-apoptotic family gene. As a result, using our experiment microarray, we selected genes that were associated with Bax in microarray data, and eventually selected the Tgfbr2 gene. Expression of the Tgfbr2 gene was decreased by methamphetamine in Bax knockout mice, and the gene was overexpressed in Bax wild-type mice. Additionally, we confirmed that Creb, FosB, and c-Fos were related to rewarding effect and Bax using immunohistochemistry.

• The Journal of Korean Medicine
• /
• v.20 no.1 s.37
• /
• pp.91-105
• /
• 1999

The effects of Yinjin and Yinjinsaryongsangagambang on a DNA damaging agent, etoposide-induced apoptosis, cell viability, cell cycle progression, and mRNA expression of apoptosis-related genes of human hepatocyte cell line HepG2 were investigated using tryphan blue exclusion assay, MTT assay, flow cytometry, immunocytometric analysis of PCNA, and quantitative RT-PCR analysis. MTT assay showed that Yinjin and Yinjinsaryongsangagambang increases cellular viability of HepG2 cells in a dosage-dependent manner. Stimulation of cell cycle progression by Yinjin or Yinjinsaryongsangagambang was detected by flow cytometric analysis of the DNA content and immunocytometric analysis of PCNA expression. A significant reduction of a DNA-damaging agent, etoposide-induced apoptosis were found in both Yinjin and Yinjinsaryongsangagambang-treated cells in dosage-dependent manner. In overall, 3-fold reduction of apoptosis was recognized in $10.0\;{\mu}g/ml$ of Yinjin or Yinjinsaryongsangagambang-treated cells compared to untreated cells. Although the difference is not significant, Yinjinsaryongsangagambang showed slightly higher effect on the inhibition of apoptosis than Yinjin. From flow cytometric analysis of apoptosis, while 39.9% of untreated cells showed etoposide-induced apoptotic cell death, only 19.6% or 17.4% of Yinjin or Yinjinsaryongsangagambang-treated cells were fond at apoptotic sub G1 phase, respectively. Interestingly, strong induction of Gadd45-mRNA was observed from Yinjin or Yinjinsaryongsangagambang-treated cells. However, no changes in expression levels of p53 and Waf1 were detected, demonstrating that induction of Gadd45 mRNA expression by Yinjin or Yinjinsaryongsangagambang occurs by p53-independent mechanism. Marked mRNA inductions of two apoptosis-inhibiting genes, Bcl-2 and Bcl- XL, were found in both Yinjin or Yinjinsaryongsangagambang-treated HepG2 cells while no changes was detected in expression levels of an apoptosis-promoting gene, Bax.

• The Journal of Korean Obstetrics and Gynecology
• /
• v.24 no.4
• /
• pp.31-49
• /
• 2011

Objectives: This study was performed to assess the protective effect of Polygonum multiflorum(PM) on UVB-irradiated HaCaT Keratinocytes damage. Methods: The protective effects of Polygonum multiflorum(PM) were determined by UVB-irradiated HaCaT assay. We assessed protective effects of Polygonum multiflorum(PM) on LDH release and nitrite production from HaCaT. COX-2, Bcl-2, Bax, $TNF{\alpha}$, c-jun, c-fos, NF-${\kappa}B$, iNOS, Bcl-xL gene expression were determined in HaCaT using real-time PCR method. Results: 1. PM inhibited LDH Release in UVB-irradiated HaCaT Keratinocytes. 2. PM inhibited Nitrite Production in UVB-irradiated HaCaT Keratinocytes. 3. PM suppressed the Gene Expression of COX-2 in UVB-irradiated HaCaT Keratinocytes. 4. PM increased the Gene Expression of Bcl-2 in UVB-irradiated HaCaT Keratinocytes. 5. PM didn't increase the Gene Expression of Bax in UVB-irradiated HaCaT Keratinocytes. 6. PM suppressed the Gene Expression of $TNF{\alpha}$ in UVB-irradiated HaCaT Keratinocytes. 7. PM suppressed the Gene Expression of c-jun in UVB-irradiated HaCaT Keratinocytes. 8. PM suppressed the Gene Expression of c-fos in UVB-irradiated HaCaT Keratinocytes. 9. PM suppressed the Gene Expression of NF-${\kappa}B$ in UVB-irradiated HaCaT Keratinocytes. 10. PM suppressed the Gene Expression of i-NOS in UVB-irradiated HaCaT Keratinocytes. 11. PM didn't increase the Gene Expression of Bcl-xL in UVB-irradiated HaCaT Keratinocytes Conclusions: In conclusion, these results suggest that PM inhibited the cell damage in UVB-irradiated HaCaT.

• Korean Journal of Food Science and Technology
• /
• v.53 no.1
• /
• pp.34-39
• /
• 2021

This study investigated the anti-cancer effects of Salicornia herbacea L. fractions in human ovarian cancer cells (OVCAR-3). S. herbacea powder was extracted with 95% EtOH and sequentially fractionated with hexane, dichloromethane (DCM), ethyl acetate, butanol, and H2O. Further, the growth inhibitory effects of the six fractions were determined using the MTS assay. The DCM fraction dramatically decreased cell viability. Similarly, the cell cycle was arrested at the subG1 phase in DCM-treated cells. To confirm apoptosis, the cells were stained with annexin V/FITC-PI solution. Total, early, and late apoptotic cells were significantly increased in the DCM fraction. The mRNA expression of Bcl-2 was reduced, whereas the pro-apoptotic factors Bax and Bak were increased in DCM fraction-treated cells. These results indicated that the DCM fraction in S. herbacea exhibited strong apoptotic effects through the p53-dependent signaling pathway.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.30 no.3
• /
• pp.150-156
• /
• 2016

Extract of Broussometia kazinoki Rhizodermatis has been traditionally used for geopoong, diuresis, hwalhyeol. In the present study, the apoptotic effect of methanol extract of Broussometia kazinoki (MBK) were investigated. Cell viability of A549 cells was measured by MTT assay. Apoptosis-related protein and MAPK protein levels were measured by Western blot. Chromatin condensation of A549 cells was stained with DAPI. MBK inhibited cell proliferation of A549 cell. Based on DAPI staining, MBK-treated cells manifested nuclear shrinkage, condensation and fragmentation. Treatment of A549 cells with MBK resulted in activation of the caspase-3, -8, -9 and cleavage of poly ADP-ribose polymerase (PARP). In the upstream, MBK increased the expressions Bax and Bak, decreased the expression of Bcl-2, and augmented the Bax/Bcl-2 ratio. MBK-induced apoptosis was accompanied by sustained phosphorylation of JNK, p38 MAPK and apoptosis signal-regulating kinase (ASK)-1. These results suggest that MBK induced apoptosis in A549 cells through Bcl-2 family protein-mediated mitochondria/caspase-3 dependent pathway. In addition, MBK increased the activation of ASK-1, which are critical upsteam signals for JNK/p38 MAPK activation in A549 cancer cells.