• Biomedical Science Letters
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• v.11 no.3
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• pp.281-287
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• 2005

Although actinomycin D (AMD) is known to induce apoptotic cell death to various cell lines, the mechanism of apoptosis induced by AMD is still unclear. Understanding this mechanism may improve its therapeutic efficacy. The present study has been performed to elucidate expression of Bcl-2 and Caspase-3 proteins related to apoptosis in human leukemia K-562 cells. Five different assays were performed in this study; DNA fragmentation analysis by agarose gel electrophoresis, quantitative assay of fragmented DNA, morphological assessment of apoptotic cells, quantification of apoptosis by annexin V (AV) and propidium iodide (PI) staning, and expression of Bcl-2 and Caspase-3 proteins by the western blot analysis. The number of apoptotic cells and amount of fragmented DNA in this cell line treated with AMD was increased at 6 hour. DNA ladder pattern was also appeared at 6 hour. The expression of Bcl-2 was decreased, and disappeared from 12 hours after AMD treatment. Precursor of Caspase-3 was degraded, and 20 kDa cleavage products were detected. These results suggest that AMD induced apoptosis of K-562 cells is Caspase-3-dependent fashion, and this apoptosis is related to the degradation of Bcl-2 proteins.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.4
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• pp.955-959
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• 2005

We previously demonstrated that the methylene chloride fraction of Scutellaria barbata suppessed human leukemic U937 cell proliferation by inducing apoptosis. In the present study, we have isolated luteolin from Scutellaria barbata and evaluated its apoptotic mechanism in Lewis lung carcinoma cells. Luteolin inhibited the proliferation of Lewis lung carcinoma cells in a concentration-dependent manner. Luteolin effectively increased the portion of $sub-G_1$ DNA content (apoptotic portion) and apoptotic Annexin-V positive cells in a concentration-dependent manner by FACS analysis. Caspase 9 and caspase 3 were activated and PARP was effectively cleaved by luteolin. It also increased the ratio of Bax to Bcl-2 through the decrease of Bcl-2 expression by Western blotting and reduced mitochondrial membrane potential following TMRE staining. These results suggest that luteolin can induce apoptosis through the mitochondrial mediated pathway.

• Nutrition Research and Practice
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• v.10 no.4
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• pp.393-397
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• 2016

BACKGROUND/OBJECFTIVES: Artemisinin, a natural product isolated from Gaeddongssuk (artemisia annua L.) and its main active derivative, dihydroartemisinin (DHA), have long been used as antimalarial drugs. Recent studies reported that artemisinin is efficacious for curing diseases, including cancers, and for improving the immune system. Many researchers have shown the therapeutic effects of artemisinin on tumors such as breast cancer, liver cancer and kidney cancer, but there is still insufficient data regarding glioblastoma (GBM). Glioblastoma accounts for 12-15% of brain cancer, and the median survival is less than a year, despite medical treatments such as surgery, radiation therapy, and chemotherapy. In this study, we investigated the anti-cancer effects of DHA and transferrin against glioblastoma (glioblastoma multiforme, GBM). MATERIALS/METHODS: This study was performed through in vitro experiments using C6 cells. The toxicity dependence of DHA and transferrin (TF) on time and concentration was analyzed by MTT assay and cell cycle assay. Observations of cellular morphology were recorded with an optical microscope and color digital camera. The anti-cancer mechanism of DHA and TF against GBM were studied by flow cytometry with Annexin V and caspase 3/7. RESULTS: MTT assay revealed that TF enhanced the cytotoxicity of DHA against C6 cells. An Annexin V immune-precipitation assay showed that the percentages of apoptosis of cells treated with TF, DHA alone, DHA in combination with TF, and the control group were $7.15{\pm}4.15%$, $34.3{\pm}5.15%$, $66.42{\pm}5.98%$, and $1.2{\pm}0.15%$, respectively. The results of the Annexin V assay were consistent with those of the MTT assay. DHA induced apoptosis in C6 cells through DNA damage, and TF enhanced the effects of DHA. CONCLUSION: The results of this study demonstrated that DHA, the derivative of the active ingredient in Gaeddongssuk, is effective against GBM, apparently via inhibition of cancer cell proliferation by a pharmacological effect. The role of transferrin as an allosteric activator in the GBM therapeutic efficacy of DHA was also confirmed.

• Biomedical Science Letters
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• v.26 no.4
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• pp.249-255
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• 2020

Radiation therapy is one of the primary options for the treatment of malignant tumors. Even though it is an effective anti-cancer treatment, it can cause serious complications owing to radiation-induced damage to the normal tissue around the tumor. It was recently reported that normal stem cell response to the genotoxic stress of ionizing radiation can boost the therapeutic effectiveness of radiation by repairing damaged cells. Therefore, we focused on annexin A-1 (ANXA1), one of the genes induced by low-dose irradiation, and assessed whether it can protect adipose-derived stem cells (ADSCs) against oxidative stress-induced damage caused by low-dose irradiation and improve effectively cell survival. After confirming ANXA1 expression in ADSCs transfected with an ANXA1 expression vector, exposure to hydrogen peroxide (H2O2) was used to mimic cellular damage induced by a chronic oxidative environment to assess cell survival under oxidative conditions. ANXA1-transfected ADSCs demonstrated that increased viability compared with un-transfected cells and exhibited enhanced anti-oxidative properties. Taken together, these results suggest that ANXA1 could be used as a potential therapeutic target to improve the survival of stem cells after low-dose radiation treatment.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.4
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• pp.1663-1666
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• 2012

Background and Objective: Protein expression in colon and rectal cancer (CRC) and paired normal tissues was examined by two-dimensional gel electrophoresis (2-DE) to identify differentially expressed proteins. Materials and Methods: Five fresh colorectal cancer and paired adjacent normal tissues were obtained and differentially expressed protein spots were determined using PDQuest software, with identification on the basis of MALDI-TOF mass spectra. Results: Compared with normal colorectal mucosa, protein abnormal expression of 65 spots varying more than 1.5 times were found in 2-DE gels from colorectal cancer samples (P<0.05); forty-two proteins were up-regulated and 23 were down-regulated; twelve protein spots were identified using mass spectrometry, of which 8 were up-regulated, includimng HSPB1and Annexin A4, while 4 were down-regulated, the results being consistent with Western blot findings. Conclusions: Two-dimensional electrophoresis reference maps for CRC tissues and adjacent normal mucosa (NMC) were established and 12 differentially expressed proteins were identified. Up-regulated HSPB1 and Annexin A4 may play many important roles in the pathogenesis of colorectal cancer.

• The Journal of Korean Obstetrics and Gynecology
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• v.28 no.2
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• pp.15-25
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• 2015

Objectives : In this study, we investigated the anti-proliferative and apoptosis inducing effect of water extract of Chelidonium majus (CM) on human breast cancer cell MDA-MB-231. Methods : The MTT assay was used to assess cell proliferation. The expression of apoptosis related gene was assessed by quantitative Real-time PCR. Cell apoptosis detected by flow cytometry using Annexin-V/PI staining. Results : Our data revealed that CM inhibited the cell growth in a dose dependent manner (0, 62.5, 125, 250, 500 μg/ml). CM increased mRNA expression of pro-apoptotic genes Bax, caspase-3, and caspase-9. Annexin-V/PI staining assays revealed that apoptosis-induced cell death increased in a dose-dependent manner in cells. Conclusions : CM induces cell death in MDA-MB-231 human breast cancer cell and shows potentials for use in cancer therapy as apoptosis-inducing agent.

• Asia-pacific Journal of Multimedia Services Convergent with Art, Humanities, and Sociology
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• v.7 no.1
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• pp.395-405
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• 2017

To confirm potential anti-cancer activities of ethylacetate (EtOAc) fraction from Orostachys japonicus on the A549 human lung cancer cells, this study examined. As a result of conducting MTS assay for measuring cell viability, the EtOAc fraction inhibited the proliferation of A549 cells in a dose-dependent manner. To investigate whether the inhibiting A549 cell viability was caused by apoptosis, this study analyzed chromatin condensation in A549 cells using DAPI staining. The morphological changes such as the formation of nuclear condensation were formed in a dose-dependent manner. Also, this study performed Annexin V-FITC staining for detecting phosphatidylinositol (PS). As a result of Annexin V-FITC staining to investigate level of early and late apoptosis, the apoptosis level treated with EtOAc fraction was higher than that of control. RT-PCR was performed to study the correlation between G2/M cell cycle arrest and cell cycle control genes. The anti-cancer activity of EtOAc fraction was accompanied by inhibition of CDK1, 4, cyclin B1 and D1 mRNA. This study also examined the expression of various marker proteins: p53, Bax, Bcl-2 and pro-caspase 3. Western blotting revealed that p53 and Bax proteins were up-regulated, and Bcl-2 and pro-caspase 3 proteins down-regulated in a time and dose-dependent manner.

• 대한핵의학회:학술대회논문집
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• 2004.11a
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• pp.408.2-408.2
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• 2004

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.27 no.4
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• pp.158-176
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• 2014

Purpose : The purpose of this study is to investigate the effect of ARTEMISIAE CAPILLARIS HERBA and COPTIDIS RHIZOMA Extracts on cell death in pancreatic cancer cells. Method : Human-derived pancreatic cancer cell line, MIA PaCa-2 cells were treated by Prescription A with various concentrations and the cytotoxicity was determined by MTT assay. To investigate the effects of Prescription A on pancreatic cancer cells, the staining of Annexin V/PI, cell cycle arrest, nuclear chromatin condensation and the production of reactive oxygen species (ROS) were examined. The effect of Prescription A's effective components, ARTEMISIAE CAPILLARIS HERBA and COPTIDIS RHIZOMA Extracts on cell death were also observed. Results : The viability of MIA PaCa-2 cells treated with Prescription A were decreased in a dose dependent manner. Prescription A induced cell death in MIA PaCa-2 cells as shown by result of Annexin V/PI double staining, chromatin condensation and cell cycle arrest. In addition, production of ROS was increased by Prescription A treatment, suggesting that ROS induced by Prescription A mediated cell death. Furthermore, Prescription A's effective components, ARTEMISIAE CAPILLARIS HERBA and COPTIDIS RHIZOMA Extracts were also induced apoptosis of MIA PaCa-2 cells through ROS production. Conclusion : These results suggest that Prescription A's effective components, ARTEMISIAE CAPILLARIS HERBA and COPTIDIS RHIZOMA Extracts induced death of MIA PaCa-2 through ROS production.

• Reproductive and Developmental Biology
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• v.31 no.3
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• pp.207-213
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• 2007

This study was carried out to examine gene expression altered in endometrium of Korean cattle (Hanwoo) with endometritis using microarray. In this study, 4,560 diferentially expressed genes (DEGs) were identified in the endometrium of Hanwoo. Of 4,560 DEGs, 2,026 genes were up-regulated, while 2,536 genes were down-regulated in endometritis. Of them, top 10 regulated genes were listed. Filamin A, pancreatic anionic trypsinogen, Rho GDP dissociation inhibitor alpha, collagen type VI alpha 1, butyrate response factor 2, aggrecanses-2, annexin 14, aminopeptidease A, orphan transporter v7-3, and epithelial stromal interaction 1 were up-regulated, while MHC class II antigen, integrin-binding sialoprotein, uterine milk protein precursor, down-regulated in colon cancer 1, glycoprotein 330, dickkopf-1, cfh protein, $Ca^{2+}-dependent$ secretion activator, UL16 binding protein 3, and proenkephalin were down-regulated in the endometritis. Our results suggest that these genes could be useful biomarkers for diagnosis Hanwoo's endometritis.