• Journal of Life Science
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• v.23 no.9
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• pp.1109-1117
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• 2013

Induction of G1 Arrest by Methanol Extract of Lycopus lucidus in Human Lung Adenocarcinoma A549 Cells Lycopus lucidus, a herbaceous perennial, is used as a traditional remedy in East Asia, including China and Korea. It has been reported that L. lucidus has anti-allergic effects, inhibitory effects on cholesterol acyltransferase in high glucose-induced vascular inflammation, and anti-proliferative effects in human breast cancer cells. However, the molecular mechanisms of the anti-cancer effects of L. lucidus have not yet been fully determined. In this study, we evaluated the anti-cancer effect and the mechanism of action of L. lucidus in human lung adenocarcinoma A549 cells using methanol extracts of L. lucidus (MELL). MELL treatment showed cytotoxic activity in a dose-dependent manner and induced G1 arrest in A549 cells. The induction of G1 arrest by MELL was associated with the up-regulation of phospho-CHK2 and the down-regulation of Cdc25A phosphatase. In addition, MELL treatment induced decreased expression of G1/S transition-related proteins, including CDK2, CDK4, CDK6, cyclin D1 and cyclin E. MELL also regulated the mRNA expression of CDK2 and cyclin E. On the other hand, the expression of p53 and the cyclin-dependent kinase inhibitor p21 was not induced by MELL. Collectively, these results suggest that MELL may exert an anti-cancer effect by cell cycle arrest at G1 phase through the ATM/CHK2/Cdc25A/CDK2 pathway in A549 cells.

• Biomolecules & Therapeutics
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• v.26 no.3
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• pp.328-334
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• 2018

Because of the unsatisfactory treatment options for breast cancer (BC), there is a need to develop novel therapeutic approaches for this malignancy. One such strategy is chemotherapy using non-toxic dietary substances and botanical products. Studies have shown that Panduratin A (PA) possesses many health benefits, including anti-inflammatory, anti-bacterial, anti-oxidant and anticancer activities. In the present study, we provide evidence that PA treatment of MCF-7 BC cells resulted in a time- and dose-dependent inhibition of cell growth with an $IC_{50}$ of $15{\mu}M$ and no to little effect on normal human MCF-10A breast cells. To define the mechanism of these anti-proliferative effects of PA, we determined its effect critical molecular events known to regulate the cell cycle and apoptotic machinery. Immunofluorescence and flow cytometric analysis of Annexin V-FITC staining provided evidence for the induction of apoptosis. PA treatment of BC cells resulted in increased activity/expression of mitochondrial cytochrome C, caspases 7, 8 and 9 with a significant increase in the Bax:Bcl-2 ratio, suggesting the involvement of a mitochondrial-dependent apoptotic pathway. Furthermore, cell cycle analysis using flow cytometry showed that PA treatment of cells resulted in G0/G1 arrest in a dose-dependent manner. Immunoblot analysis data revealed that, in MCF-7 cell lines, PA treatment resulted in the dose-dependent (i) induction of $p21^{WAF1/Cip1}$ and p27Kip1, (ii) downregulation of Cyclin dependent kinase (CDK) 4 and (iii) decrease in cyclin D1. These findings suggest that PA may be an effective therapeutic agent against BC.

• Natural Product Sciences
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• v.18 no.1
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• pp.16-21
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• 2012

Honokiol, a naturally occurring neolignan mainly found in Magnolia species, has been shown to have the anti-angiogenic, anti-invasive and cancer chemopreventive activities, but the molecular mechanism of actions has not been fully elucidated yet. In the present study, we investigated the effect of honokiol on the growth inhibitory activity in cultured SNU-638 human gastric cancer cells. We found that honokiol exerted potent antiproliferative activity against SNU-638 cells. Honokiol also arrested the cell cycle progression at the G0/G1 phase and induced the apoptotic cell death in a concentration-dependent manner. The cell cycle arrest was well correlated with the downregulation of Rb, cyclin D1, cyclin A, cyclin E, and CDK4 expression, and the induction of cyclin-dependent kinase inhibitor p27. The increase of sub-G1 peak by honokiol was closely related to the induction of apoptosis, which was evidenced by the induction of DNA fragmentation, the cleavage of poly(ADPribose) polymerase, and the sequential activation of caspase cascade. These findings suggest the cell cycle arrest and induction of apoptosis might be one possible mechanism of actions for the anti-proliferative activity of honokiol in human gastric cancer cell.

• Korean Journal of Head & Neck Oncology
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• v.27 no.1
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• pp.59-65
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• 2011

Background and Objectives : Papillary thyroid carcinoma(PTC) frequently metastasize to the regional neck, however, lateral neck lymph node metastasis is less common. The aim of this study is to investigate clinical and immunohistochemical features of PTC with lateral LN metastasis, and determine the predictive factors for lateral LN metastases. Material and Methods : We undertook a retrospective study of 83 patients treated between January 2007 and December 2009 for PTC by thyroidectomy with or without lateral neck dissection. The following criteria were used to study the clinical predictive value of lateral LN. metastases : sex, age, tumor size, multifocality, extracapsular spread(ECS) and lymphovascular emboli. Immunohistochemical staining for VEGF-A, VEGF-C, Bax, Bcl-2, Cyclin D1, Cyclin E, $p27^{kip1}$ and $p57^{kip2}$ was performed, and quantified blindly by three pathologists who had no clinical information of the patients. Immunohistochemical expression was scored as high(>50% of cells stained) or low(0-49%). Results : With use of univariate and multivariate analysis, tumor size(>2cm) and ECS were independent correlates of lateral LN metastasis in PTC. Expression of VEGF-C, Bax, and Cyclin D1 in the PTC with lateral LN metastasis was scored higher than in PTC without lateral LN metastasis(p<0.05). Conclusion : The important risk factors for lateral LN metastasis in PTC are primary tumor size and the presence of ECS. And expression of VEGF-C, Bax and cyclin D1 may be considered of lateral LN metastatic potential in PTC.

• Journal of Microbiology and Biotechnology
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• v.22 no.12
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• pp.1790-1794
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• 2012

Recently, it has been suggested that $p27^{Kip1}$, the cell cycle regulatory protein, plays a pivotal role in the progression of normal differentiation in murine embryonic stem (mES) cells. In the current study, we investigated the role of $p27^{Kip1}$ in the regulation of differentiation and apoptotic induction using Western blotting, quantitative real-time RT-PCR, and small interfering RNA (siRNA) assays and confocal laser scanning microscopic analysis of H9 human ES (hES) cells and H9-derived embryoid bodies (EBs) grown for 10 ($EB_{10}$) and 20 days ($EB_{20}$). Our results demonstrate that the proteins $p27^{Kip1}$ and cyclin D3 are strongly associated with cellular differentiation, and, for the first time, show that up-regulation of $p27^{Kip1}$ protects hES cells from inducing differentiation-associated and caspase 3-dependent apoptosis.

• Journal of Periodontal and Implant Science
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• v.33 no.3
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• pp.415-437
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• 2003

Ginseng Radix(GR) had been used widely from oriental medicine and the effects of it have been investigated by many researchers. The purpose of present study was to investigate the effects of GR on the cell cycle progression and its molecular mechanism in human fetal osteoblast. The results were as follows. Increased cell proliferation was observed in cells exposed to 100 ng/ml, 10 ng/ml of GR-1 at 12 hours and 24 hours, 1 ${\mu}g$/ml of GR-1 at 48 hours, and 100 ${\mu}g$/ml, 10 ${\mu}g$/ml of GK-2 at 12 hours, all treatment groups of GR-2 at 24 hours(p<0.05). S phase and G1 phase was increased in the group of treated with 100 ng/ml of GR-1, with 10 ${\mu}g$/ml and 1 ${\mu}g$/ml of GR-2, with 100 ${\mu}g$/ ml and 10 ${\mu}g$/ml of GR-3 in the cell cycle analysis. The cell cycle regulation protein levels of Cyclin D1, Cyclin E, CDK 2. CDK 4 and CDK 6 were increased in the group of treated with 1 ${\mu}g$/ml and 100 ng/ml of GR-1, with 10 ${\mu}g$/ml and 1 ${\mu}g$/ml of GR-2, with 100 ${\mu}g$/ ml and 10 ${\mu}g$/ml of GR-3. On the other hand, p21 was decreased in the treatment group with 1 ${\mu}g$/ml and 100 ng/ml of GR-1, with 10 ${\mu}g$/ml and 1 ${\mu}g$/ml of GR-2, 10 ${\mu}g$/ml of GR-3, and p53 and p16 was decreased in the treatment group with 100 ng/ml of GR-1, 100 ${\mu}g$/ml and GR-3 and pRb was decreased in the all treatment groups except 1 ${\mu}g$/ml of GR-1. These results suggested that GR increases the cell proliferation and the cell cycle progression in human fetal osteoblast, which is linked to increased cell cycle regulation protein levels of Cyclin D1 , Cyclin E, CDK 2, CDK 4, CDK 6 and decreased cell cycle regulation protein levels of p21, pRb.

• Journal of Life Science
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• v.19 no.7
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• pp.871-877
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• 2009

We investigated the effects of sodium butyrate, a histone deacetylase inhibitor, on the cell cycle progression in human monocytic leukemia U937 cells. Exposure of U937 cells to sodium butyrate resulted in growth inhibition, G1 arrest of the cell cycle and induction of apoptosis in a dose-dependent manner as measured by MTT assay and flow cytometry analysis. The increase in G1 arrest was associated with the down-regulation in cyclin D1, E, A, cyclin-dependent kinase (Cdk) 4 and 6 expression, and up-regulation of Cdk inhibitors such as p21 and p27. Sodium butyrate treatment also inhibited the phosphorylation of retinoblastoma protein (pRB) and p130, however, the levels of transcription factors E2F-1 and E2F-4 were not markedly modulated. Furthermore, the down-regulation of phosphorylation of pRB and p130 by this compound was associated with enhanced binding of pRB and E2F-1, as well as p130 and E2F-4, respectively. Overall, the present results demonstrate a combined mechanism involving the inhibition of pRBjp130 phosphorylation and induction of Cdk inhibitors as targets for sodium butyrate that may explain some of its anti-cancer effects in U937 cells.

• The Journal of Korean Medicine
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• v.36 no.4
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• pp.19-34
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• 2015

Objectives: This study was performed to investigate the anti-tumor effect of O. japonicus extracts on intrahepatic cholangiocarcinoma cell line SNU-1079. Methods: Cholangiocarcinoma SNU-1079 cells were treated with various concentrations of O. japonicus DW and EtOH extracts ($0-300{\mu}g/ml$) for 24, 48 or 72 h. Cell viability was evaluated through a PMS/MTS assay, and the apoptosis rate was examined through ELISA assay and flow cytometry analysis. The mRNA expression of apoptosis- and cell cycle progression-related genes (Bcl-2, Mcl-1, Bax, Survivin, Cyclin D1, and p21) was evaluated using real-time PCR, and the caspase activity was examined using immunoblot analysis. Results: O. japonicus extracts inhibited cell proliferation and increased apoptosis rate in both ELISA assay and flow cytometry analysis. O. japonicus extracts decreased Bcl-2, Mcl-1, Survivin, and Cyclin D1 mRNA expression and increased Bax mRNA level. O. japonicus extracts also increased Caspase-3 activation. Overall, O. japonicus DW extracts were more effective than EtOH extracts. Conclusions: O. japonicus inhibited cell proliferation and induced apoptosis in SNU-1079 cells via mitochondria -mediated intrinsic pathway, which leads to Caspase-3 activation. The results indicate that O. japonicus is a potential therapeutic herb with anti-tumor effect against intrahepatic cholangiocarcinoma.

• Journal of Applied Biological Chemistry
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• v.66
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• pp.379-387
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• 2023

Dog owners seek treatment when their pets develop cancer. IMMUNIES is traditional herbal medicine-based figment made of 10 natural herbs, designed to maintain host immune function. The major component of IMMUNIES is Dendropanax morbiferus. This clinical pilot study monitored the toxicity and efficacy of IMMUNIES. Four senile dogs with spontaneously occurring mammary and liver cancers were enrolled in this study and treated orally daily for 3 months, and their blood/urine biochemical profiles were examined each month. IMMUNIES was well tolerated during the treatment period. Blood urea nitrogen, creatinine, alanine aminotransferase, alkaline phosphatase, and C-reactive protein levels decreased in all four dogs, whereas red blood cells and hematocrit were within the normal range. IMMUNIES also changed the expression of several molecular targets in the anticancer pathway, such as pro-NAG-1, p53, and cyclin D1. Although the tumors did not completely respond to IMMUNIES, the biochemical profiles and clinical examination showed a stabilized cancer status for 3 months. Thus, IMMUNIES was found to be safe and well-tolerated in the dosage range tested and exhibited cancer antiproliferative activity in canine cancer. Future studies should address other potential benefits of IMMUNIES, including correlative assessments of immune function, quality of life, and owner satisfaction.

• Journal of Pharmacopuncture
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• v.7 no.2
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• pp.5-17
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• 2004

Objective : In order to measure the efficacy of cultivated wild ginseng distilled herbal acupuncture by concentration level, we've treated A549 human lung cancer lines with different concentrations of cultivated wild ginseng distilled herbal acupuncture and examined mRNA and proteins which take parts in apoptosis. Methods : A549 human lung cancer lines were treated with various concentration levels of cultivated wild ginseng distilled herbal acupuncture and cell toxicity was carefully examined. From the analysis of DNA fragmentation, RT-PCR and Western blot, manifestation of mRNA and proteins which are associated with apoptosis were inspected. Results : The following results were obtained on apoptosis of A549 human lung cancer lines after administering various concentration levels of cultivated wild ginseng distilled herbal acupuncture. 1. Measuring cell toxicity of lung cancer cells, strong cell toxicity was detected at high concentration level(1000ul, 1200ul), but no consistent concentration dependent reliance was detected. 2. Through DNA fragmentation, we were able to confirm cell destruction in all groups. 3. Experiment groups treated with cultivated wild ginseng distilled herbal acupuncture showed inhibition of Bcl-2 and COX-2 at mRNA and Protein level, whileas increase of Bax was shown. 4. Manifestation of p21, p53, Cyclin E, and Cyclin Dl were confirmed in all groups. 5. Extrication of Cytochrome C was detected at all groups, as well as increased activity of the enzyme caspase-3 and caspase-9, and PARP fragmentation were confirmed. Conclusion : According to the results, we can carefully deduce cell destruction of A549 human lung cancer lines were induced by Apoptosis. At the fixed level, cultivated wild ginseng distilled herbal acupuncture showed decrease of Bcl-2 and COX-2, as well as increase of Bax. Since cultivated wild ginseng distilled herbal acupuncture increases manifestation of p21, p53, Cyclin E, and Cyclin Dl, it affects cellular cycle and through these phenomena, we can consider extrication of Cytochrome C, increase of caspase, and PARP fragmentation are the results.