• Asian Pacific Journal of Cancer Prevention
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• 제15권3호
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• pp.1163-1169
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• 2014

Astragalus, a commonly used traditional Chinese medicine, has exhibited antitumor actions in patients. In this study, in vitro and in vivo antitumor effects of astragalus and synergistic antitumor efficacy in combination with pterostilbene were investigated. Melanoma cells were treated with pterostilbene (Pt), graduated doses of astragalus injection (AI), or these in combination. Cell viability was measured using a MTT assay. Released nucleosomes and caspase activity were measured using enzyme-linked immunosorbent assay. Growth inhibition in vitro and in vivo was also assessed. Analysis of variance and t tests were used for statistical analysis. Significant reduction (p<0.05) in cellular proliferation were observed with AI and AI-Pt in a time- and concentration-dependent manner. Apoptosis and caspase-3/7 activity were significantly increased by AI and AI-Pt treatment (p<0.05). In vivo, AI inhibited melanoma tumor growth, with inhibition rates ranging from 36.5 to 62.3%, by inducing apoptosis via up-regulation Bax expression and the Bax/Bcl-2 ratio and down-regulating Bcl-2 expression. AI significantly inhibits the growth of melanoma in vitro and in vivo by inducing apoptosis. These data suggest that combined treatment of astragalus with pterostilbene enhances antitumor efficacy.

• 대한한의학회지
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• 제25권4호
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• pp.95-107
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• 2004

Objective : The inappropriate or excessive apoptosis has been known to be associated with neurodegenerative disorders including intracranial hemorrhage(ICH). Paeoniae radix, in traditional Korean medicine, has played its role as blood­nourisher and yin-astringent. In the present study, the effect of Paeoniae radix on the inhibition of neurodegeneration in the brain of rats after artificial ICH and on the resulting apoptosis was investigated. Methods : 30 rats were divided into 6 equal groups ; the sham-operation group, the hemorrhage-induction group, the hemorrhage-induction with 10, 50, 100, and 200 mg/kg Paeoniae radix-treated group, respectively. Stereotactic surgery was performed and collagenase was infused to induce ICH in the region of CA1 of hippocampus of rats. The sham group took only saline infusion. For 7 days after the surgery, 4 testing groups had intraperitoneal injections of Paeoniae radix extract. The step-down inhibitory avoidance task, measurement of neurodegeneration degree in the CA1 region of the hippocampus, and detection of caspase-3 and newly generated cells in the dentate gyrus were done after animal sacrifice. Results : Rats receiving Paeoniae radix extract showed increased latency time in the inhibitory avoidance task. The extension of neuron-deprived areas in the CA1 region was significantly suppressed in the Paeonia treated groups. Also expressions of caspase-3 in the CA1 region and cortex were significantly inhibited in the Paeonia treated groups. The cell proliferation was evaluated by means of BrdU methods and proved to be decreased in the Paeonia treated groups. Conclusion : These results suggest that Paeoniae radix has potential to suppress short-tenn memory loss after devastating neurologic accidents. Also it was proved that Paeoniae radix has a neuroprotective effect and alleviates central nervous complications following intracerebral hemorrhage. Furthermore, it may imply that this medicinal plant can be widely used for vascular dementia and other neurodegenerative disorders.

• Asian Pacific Journal of Cancer Prevention
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• 제13권11호
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• pp.5887-5895
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• 2012

Background: To determine the effect of essential oil obtained from a traditionally used medicinal plant Tridax procumbens L, on lung metastasis developed by B16F-10 melanoma cells in C57BL/6 mice. Materials and Methods: Parameters studied were toxicity, lung tumor nodule count, histopathological features, tumor directed capillary vessel formation, apoptosis and expression levels of $P^{53}$ and caspase-3 proteins. Results: In vitro the MTT assay showed cytotoxicity was found to be high as 70.2% of cancer cell death within 24hrs for $50{\mu}g$. In vivo oil treatment significantly inhibited tumor nodule formation by 71.7% when compared with untreated mice. Formation of tumor directed new blood vessels was also found to be inhibited to about 39.5%. TUNEL assays also demonstrated a significant increase in the number of apoptotic positive cells after the treatment. $P^{53}$ and caspase-3 expression was also found to be greater in the essential oil treated group than the normal and cancer group. Conclusions: The present investigation showed significant effects of the essential oil of Tridax procumbens L in preventing lung metastasis by B16F-10 cell line in C57BL/6 mice. Its specific preventive effect on tumor directed angiogenesis and inducing effect on apoptosis warrant further studies at the molecular level to validate the significance of Tridax procumbens L for anticancer therapy.

• International Journal of Oral Biology
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• 제36권3호
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• pp.155-162
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• 2011

Eugenol (4-allyl-2-methoxyphenol) is a naturally occurring phenolic compound that is widely used in dentistry as a component of zinc oxide eugenol cement that is commonly applied to the mouth environment. Cisplatin is one of the most potent known anticancer agents and shows significant clinical activity against a variety of solid tumors. This study was undertaken to investigate the synergistic apoptotic effects of co-treatments with eugenol and cisplatin on human melanoma (G361) cells. To investigate whether this co-treatment efficiently reduces the viability of G361 cells compared with each single treatment, an MTT assay was conducted. The induction and augmentation of apoptosis were confirmed by DNA electrophoresis, Hoechst staining and an analysis of DNA hypoploidy. Western blot analysis and immunofluorescent staining were also performed to evaluate the expression levels and the translocation of apoptosis-related proteins following this co-treatment. Furthermore, proteasome activity and mitochondrial membrane potential (MMP) changes were also assayed. The results indicated that a co-treatment with eugenol and cisplatin induced multiple pathways and processes associated with an apoptotic response in G361 cells including nuclear condensation, DNA fragmentation, a reduction in MMP and proteasome activity, the increase and decrease of Bax and Bcl-2, a decreased DNA content, the release of cytochrome c into the cytosol, the translocation of AIF and DFF40 (CAD) into the nucleus, and the activation of caspase-9, caspase-7, caspase-3, PARP and DFF45 (ICAD). In contrast, separate treatments of 300 ${\mu}M$ eugenol or 3 ${\mu}M$ cisplatin for 24 h did not induce apoptosis. Our present data thus suggest that a combination therapy of eugenol and cisplatin is a potential treatment strategy for human melanoma.

• International Journal of Oral Biology
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• 제38권3호
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• pp.101-110
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• 2013

We investigated the synergistic apoptotic effects of co-treatments with Chios gum mastic (CGM) and eugenol on G361 human melanoma cells. An MTT assay was conducted to investigate whether this co-treatment efficiently reduces the viability of G361 cells compared with each single treatment. The induction and augmentation of apoptosis were confirmed by DNA electrophoresis, Hoechst staining, and analyses of DNA hypoploidy. Western blot analysis and immunofluorescent staining were also performed to evaluate expression and translocation of apoptosis-related proteins following CGM and eugenol co-treatment. Proteasome activity and mitochondrial membrane potential (MMP) changes were also assayed.The results indicated that the co-treatment of CGM and eugenol induces multiple pathways and processes associated with an apoptotic response in G361 cells. These include nuclear condensation, DNA fragmentation, a reduction in MMP and proteasome activity, an increase of Bax and decrease of Bcl-2, a decreased DNA content, cytochrome c release into the cytosol, the translocation of AIF and DFF40 (CAD) into the nucleus, and the activation of caspase-9, caspase-7, caspase-3, PARP and DFF45 (ICAD). In contrast, separate treatments of $40{\mu}g/ml$ CGM or $300{\mu}M$ eugenol for 24 hours did not induce apoptosis. Our present data thus suggest that a combination therapy of CGM and eugenol is a potential treatment strategy for human melanoma.

• 동의생리병리학회지
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• 제20권1호
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• pp.187-192
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• 2006

Herbal medicines have been utilized to treat a variety of diseases, including cancer. Several species of the family rubiaceae have been reported to have antitumor activity. In this study, we report the cytotoxicity and antitumor activity exhibited dy the methanol extracts prepared from Rubia radix (RRME), Uncaria gambir (UGME) and Oldenlandia diffusa (ODME) (family: Rubiaceae) against human promyleloid leukemia cell line, HL-60. The cytotoxicity of RRME (2~20 ${\mu}g/ml$), UGME (20~200 ${\mu}g/ml$) and ODME (20~200 ${\mu}g/ml$) were assessed dy the MTT reduction assay. IC50 values for RRME, UGME and ODME were 11.0, 99.5 and 106.1 ${\mu}g/ml$, respectively. When the HL-60 cells were treated with RRME (10 ${\mu}g/ml$), UGME (120 ${\mu}g/ml$) and ODME (140 ${\mu}g/ml$) for 24 h, several apoptotic characteristics such as DNA fragmentation and morphologic changes were observed. Furthermore, flow cytometric analysis was peformed to determine the percent of apoptotic cells. The poupulation of sub-G1 hypodiploid cells was increased 37.49% in RRME treatment, 12.49% in UGME treatment and 7.21% in ODME treatment compared with untreated control cells (2.64%). To further confirm apoptotic cell death, we assayed caspase-3, -8 and -9 activities in RRME, UGME and ODME-treated cells. After treatment of RRME, UGME and ODME for 12 h, caspase-3, -8 and -9 activities significantly increased.compared to untreated control cells. These results show that RRME, UGME and ODME induced apoptotic cell death in HL-60 cells and may have a possibility of potential antitumor activities.

• 대한화장품학회지
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• 제41권1호
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• pp.1-7
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• 2015

말채나무는 한국의 민간요법으로 사용되던 약재이다. 자외선은 피부의 광손상을 일으키는 것으로 알려져 있다. 본 연구에서는 자외선에 의한 손상된 세포를 회복시키기 위해서 효소처리 된 말채나무잎추출물(CWE)을 사용하였다. 섬유아세포에 UVB를 조사한 후, CWE를 처리하여 세포의 회복을 조사하였다. UVB를 조사한 섬유아세포에는 caspase-3 활성, phospho-p53, ${\gamma}H2AX$, cyclobutane pyrimidine dimers (CPDs) formation, 그리고 DNA fragmentation이 증가하게 된다. 그러나 CWE를 UVB가 조사된 섬유아세포에 12 h 처리하였을 때 caspase-3 활성, phospho-p53, ${\gamma}H2AX$, CPDs formation, 그리고 DNA fragmentation이 감소하였다. 또한 CWE은 인체첩포시험을 통해 인체피부에 자극을 유발하지 않음을 확인하였다. 이러한 결과를 종합할 때 CWE는 자외선에 대한 광보호 효과가 있는 원료로서 가능성을 가지고 있다고 판단된다.

• Journal of Ginseng Research
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• 제43권4호
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• pp.692-698
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• 2019

Background: Breast cancer is a severe disease and the second leading cause of cancer death in women worldwide. To surmount this, various diagnosis and treatment options for breast cancer have been developed. One of the most effective strategies for cancer treatment is to induce apoptosis using naturally occurring compounds. Compound K (CK) is a ginseng saponin metabolite generated by human intestinal bacteria. CK has been studied for its cardioprotective, antiinflammatory, and liver-protective effects; however, the role of CK in breast cancer is not fully understood. Methods: To investigate the anticancer effects of CK in SKBR3 and MDA-MB-231 cells, cell viability assays and flow cytometry analysis were used. In addition, the direct targets of CK anticancer activity were identified using immunoblotting analysis and overexpression experiments. Invasion, migration, and clonogenic assays were carried out to determine the effects of CK on cancer metastasis. Results: CK-induced cell apoptosis in SKBR3 cells as determined through 3-(4-5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide assays, propidium iodide (PI) and annexin V staining, and morphological changes. CK increased the cleaved forms of caspase-7, caspase-8, and caspase-9, whereas the expression of Bcl-2 was reduced by CK. In assays probing the cell survival pathway, CK activated only AKT1 and not AKT2. Moreover, CK inhibited breast cancer cell invasion, migration, and colony formation. Through regulation of AKT1 activity, CK exerts anticancer effects by inducing apoptosis. Conclusion: Our results suggest that CK could be used as a therapeutic compound for breast cancer.

• 운동영양학회지
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• 제15권4호
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• pp.173-182
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• 2011

Post-traumatic stress disorder (PTSD) is a stress-related mental disorder induced by severe external stressors such as assault, disaster or severe accident. We investigated the effects of treadmill exercise on short-term memory in relation to apoptosis and cell proliferation in the hippocampus following PTSD. Stress to the pregnant rats was induced by exposure of maternal rats to the hunting dog in an enclosed room. Exposure time was 10 min, repeated three times per day, with 1 hour interval. Exposure of maternal rats to the hunting dog was continued 7 days after pregnancy until delivery. The pregnant rats in the exercise groups were forced to run on a treadmill for 30 min once a day for the same duration of stress exposure. Step-down avoidance task for short-term memory, western blot for Bcl-2, Bax, and immunohistochemistry for caspase-3, 5-bromo-2'-deoxyuridine (BrdU), and Ki-67 were conducted. Maternal rats exposed to stress during pregnancy showed short-term memory impairment. Expressions of Bax, Bcl-2, ratio of Bax to Bcl-2, and caspase-3 in the hippocampus were increased in the PTSD rats. Cell proliferation in the hippocampal dentate gyrus was decreased in the PTSD rats. Treadmill exercise alleviated short-term memory impairment and suppressed expressions of Bax, the ratio of Bax to Bcl-2, and caspase-3. Treadmill exercise also increased cell proliferation. The present results indicate that treadmill exercise alleviated PTSD-induced short-term memory impairment by suppressing apoptotic cell death and enhancing cell proliferation in the hippocampus.

• Asian Pacific Journal of Cancer Prevention
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• 제13권7호
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• pp.3073-3076
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• 2012

Terpinen-4-ol is a terpene found in the rhizome of Plai (Zingiber montanum (Koenig) Link ex Dietr.). In this study apoptogenic activity and mechanisms of cell death induced by terpinen-4-ol were investigated in the human leukemic MOLT-4 cell line. Terpinen-4-ol exhibited cytotoxicity in MOLT-4 cells, with characteristic morphological features of apoptosis by Wright's staining. The mode of cell death was confirmed to be apoptosis by flow cytometric analysis after staining with annexin V-FITC and propidium iodide. A sub-G1 peak in DNA histograms of cell cycle assays was observed. Terpinen-4-ol induced-MOLT-4 cell apoptosis mediated through an intrinsic pathway involving the loss of mitochondrial transmembrane potential (MTP) and release of cytochrome c into the cytosol. In addition, terpinen-4-ol also induced apoptosis via an extrinsic pathway by caspase-8 activation resulting in the cleavage of cytosolic Bid. Truncated-Bid (tBid) translocated to mitochondria and activated the mitochondrial pathway in conjunction with down-regulation of Bcl-2 protein expression. Caspase-3 activity also increased. In conclusion, terpinen-4-ol can induce human leukemic MOLT-4 cell apoptosis via both intrinsic and extrinsic pathways.