• Journal of Applied Biological Chemistry
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• v.63 no.2
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• pp.147-152
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• 2020

Farrerol is a flavanone isolated from the traditional Chinese herb 'Man-shan-hong' (Rhododendron dauricum L.). Farrerol has been reported to have various bioactivities including anti-oxidant, anti-inflammation, and anti-fungal. However, anti-cancer effect of farrerol has not yet been reported in MCF-7 breast cancer cells. In the present study, we investigated the anti-cancer effect of farrerol on MCF-7 cells. Farrerol decreased viability and induced apoptosis of MCF-7 cells in a dose dependent manner. Ferrerol exhibited a significant anti-proliferation effect with a half-maximal inhibitory concentration (IC₅₀) values of 145.04±1.4 μM in MTT assay, when MCF-7 cells were treated with ferrerol for 48 h. Also, ferrerol induced apoptotic bodies of MCF-7 cells as evaluated by TUNEL assay and Annexin V/PI staining using FACS. By mechanism of action, ferrerol regulated the activation of p38 mitogen-activated protein kinase and altered the expression level of BAX, Bcl-2, and Poly ADP Ribose Polymerase in MCF-7 cells. In summary, our finding demonstrated that ferrerol has anti-cancer effect through regulating the activation and expression of apoptosis-related proteins in MCF-7 cells.

• Korean Journal of Food Science and Technology
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• v.44 no.4
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• pp.452-459
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• 2012

This study was conducted in order to investigate the effect of resveratrol on the induction of apoptosis in MDA-MB-231 breast cancer cells. The result of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl terazolium bromide (MTT) assay shows that cell viability significantly decreased in a dose and time-dependent manner. 4',6-diamidino-2-phenylindole (DAPI) staining shows significantly increased chromatin condensation in a dose and time-dependent manner. Resveratrol increased the expression of p53, cleaved-caspase-3, and cleaved-caspase-9, whereas the expression of PI3K/Akt decreased in a time-dependent manner. We investigated the in vivo tumor growth inhibitory effect of resveratrol. Tumor volume was significantly decreased in the 50 mg/kg resveratrol-administration group compared to the control group. In the 50 mg/kg treated group. Apoptosis cells were frequently observed by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assay. Immunohistochemistry staining shows increased the expression of p53, cytochrome-C, and cleaved-caspase-3 in the 50 mg/kg treated group. These results indicate that resveratrol induced apoptosis through PI3K/Akt and p53 signal pathway in MDA-MB-231 cell.

• Journal of Embryo Transfer
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• v.29 no.4
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• pp.345-350
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• 2014

We investigate the effect of L-glutathione (GSH), an antioxidant, treatment during the somatic cell nuclear transfer (SCNT) procedures on the in vitro development and DNA methylation status of bovine SCNT embryos. Bovine in vitro matured (IVM) oocytes were enucleated and electrofused with a donor cell, then activated by a combination of Ca-ionophore and 6-dimethylaminopurine. The recipient oocytes or reconstituted oocytes were treated with $50{\mu}M$ GSH during these SCNT procedures from enucleation to activation treatment. The SCNT embryos were cultured for 7 days to evaluate the in vitro development, apoptosis and DNA methylation in blastocysts. The apoptosis was measured by TUNEL assay and caspase-3 activity assay. Methylated DNA of SCNT embryos at the blastocyst stages was detected using a 5-methylcytidine (5-MeC) antibody. The developmental rate to the blastocyst stage was significantly higher (P<0.05) in GSH treatment group ($32.5{\pm}1.2%$, 78/235) than that of non-treated control SCNT embryos ($22.3{\pm}1.8%$, 50/224). TUNEL assay revealed that the numbers of apoptotic cells in GSH treatment group ($2.3{\pm}0.4%$) were significantly lower (P<0.05) than that of control ($3.8{\pm}0.6%$). Relative caspase-3 activity of GSH treated group was $0.8{\pm}0.06$ fold compared to that of control. DNA methylation status of blastocysts in GSH treatment group ($13.1{\pm}0.5$, pixels/embryo) was significantly lower (P<0.05) than that of control ($17.4{\pm}0.9$, pixels/embryo). These results suggest that antioxidant GSH treatment during SCNT procedures can improve the embryonic development and reduce the apoptosis and DNA methylation level of bovine SCNT embryos, which may enhance the nuclear reprogramming of bovine SCNT embryos.

• The Journal of Korean Medicine
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• v.31 no.3
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• pp.55-65
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• 2010

Background: Nitric oxide (NO) is a reactive free radical gas and a messenger molecule. NO has many physiological functions, but excessive NO production induces neurotoxicity. Objective: The present study investigated whether the aqueous extract of Polygala tenuifolia Willdenow possesses a protective effect on NO-induced apoptosis in human neuroblastoma cell line SK-N-MC. Method: For this study, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, 4,6-diamidino-2-phenylindole (DAPI) staining, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, DNA fragmentation assay, reverse transcription-polymerase chain reaction (RT-PCR), Western blot, and caspase-3 enzyme assay were performed. Result: Sodium nitroprusside (SNP) exposure significantly decreased the viability of cells. The cells treated with SNP exhibited several apoptotic features such as increasing of Bax expression, caspase-3 enzyme activity and inhibiting of Bcl-2 expression. On the other hand, the viability of cells pre-treated with the aqueous extract of Polygala tenuifolia Willdenow was increased dose-dependently. The cells pre-treated for 1 h with the aqueous extract of Polygala tenuifolia Willdenow followed by treatment with SNP showed a decreased occurrence of apoptotic features like decreasing Bax expressions, caspase-3 enzyme activity and increasing Bcl-2 expressions. The aqueous extract of Polygala tenuifolia Willdenow reduced apoptotic cell death in neuroblastoma cell line SK-N-MC through the inhibition of Bax-dependent caspase-3 activation and the increasing of Bcl-2 expression. Conclusion: Based on the present results, it is possible that Polygala tenuifolia Willdenow has therapeutic value for the treatment of a variety of NO-induced brain diseases.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.32 no.3
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• pp.1-12
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• 2019

Objectives: Amygdalin is abundant in the seeds of bitter almond and apricots of the Prunus genus and other rosaceous plants. Amygdalin is known to have antitussive and anticancer activities. Apoptosis, also known as programmed cell death, is an important mechanism in cancer treatment. Methods: In the present study, we investigated whether the aqueous extract of Amygdalin induces apoptotic cell death in ME-180 cervical cancer cells. For this study, 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, terminal deoxynuclotidyl transferase (TdT)-mediated dUTP nick end-labeling (TUNEL) assay, 4,6-diamidino-2-phenylindole (DAPI) staining, flow cytometric analysis, DNA fragmentation assay, Western blot, and caspase-3 enzyme assay were performed on ME-180 cervical cancer cells. Results: Through morphological and biochemical analyses, it was demonstrated that ME-180 cells treated with Amygdalin exhibit several apoptotic features. The treatment of Amygdalin increased the Bax expression and caspase-3 enzyme activity and decreased Bcl-2 expression. Here, we have shown that Amygdalin induces apoptotic cell death in ME-180 cervical cancer cells through Bax-dependent caspase-3 activation. These results suggest the possibility that Amygdalin exerts anti-tumor effect on human cervical cancer.

• The Korean Journal of Physiology and Pharmacology
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• v.5 no.2
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• pp.177-181
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• 2001

Recently, nonsteroidal anti-inflammatory drugs (NSAIDs) have been found to be useful in the chemoprevention of colon cancer. To investigate whether indomethacin, an NSAIDs, induces apoptosis and thus assess the possibility of its application in the chemoprevention of human lung cancer, we have performed MTT assay, TUNEL assay, DAPI staining, and flow cytometric analysis using human lung carcinoma cell line NCI-H1299. Through morphological and biochemical analyses, it was demonstrated that NCI-H1299 cells treated with indomethacin (0.5 mM) exhibit classical apoptotic features. These results suggest that indomethacin induces apoptosis in NCI-H1299 cells and that NSAIDs, including indomethacin, may be a useful tool for the chemoprevention of human lung cancer.

• Molecular & Cellular Toxicology
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• v.3 no.1
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• pp.31-35
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• 2007

Hesperidin, known as a flavonoid constituent of citrus, has been known to reduce the proliferation of several cancer cells. We investigated whether hesperidin-induced cell death on SNU-668, human gastric cancer cells. The cytotoxicity of hesperidin on SNU-668 cells was determined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay at the concentration of 1, 10, 50, and 100 ${\mu}M$. Cell viability by hesperidin was 53.18$\pm$2.85% of control value at 100 ${\mu}M$. The cell death by hesperidin showed apoptotic features, which were confirmed using a combination of 4, 6-diamidino-2-phenylindole (DAPI) staining and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay. In the apoptosis-regulating genes, treatment of hesperidin decreased mRNA expression of B-cell CLL/lymphoma 2 (BCL2), whereas expression of BCL2-associated X protein (BAX) was increased. The mRNA expression and the activity of caspase3 (CASP3), a major apoptotic factor, was significantly increased by hesperidin treatment. These results suggest that hesperidin could induce apoptosis through CASP3 activation on SNU-668, human gastric cancer cells.

• Natural Product Sciences
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• v.25 no.4
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• pp.298-303
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• 2019

This study investigated the cytotoxic effects and mechanism of action of asiatic acid in pancreatic cancer cell lines. First, we confirmed the cell viability of MIA PaCa-2 and PANC-1 cells after asiatic acid administration for 48 and 72 h. The viability of MIA PaCa-2 and PANC-1 cells decreased in a dose-dependent manner following asiatic acid administration. To investigate the underlying mechanism, we performed a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, annexin V assay, and western blotting. Asiatic acid induced apoptosis and autophagy through activation of AMP-activated protein kinase (AMPK) and inhibition of mammalian target of rapamycin (mTOR) in MIA PaCa-2 cells. Finally, the expression of miR-17 and miR-21, known as oncogenes in pancreatic cancer, was decreased by asiatic acid. These results indicate that asiatic acid has potential as a new therapeutic agent against pancreatic cancer.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.9
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• pp.3779-3783
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• 2015

Dietary prevention has been known to reduce breast cancer risk. Sesamin is one of the major components in sesame seeds and has been widely studied and proven to have anti-proliferation and anti-angiogenic effects on cancer cells. In this study, the influence of sesamin was tested in the human breast cancer MCF-7 cell line for cell viability (MTT assay) and cell cycling (flow cytometry). Results showed that sesamin dose-dependently (1, 10 and $50{\mu}M$) reduced the cell viability and increased LDH release and apoptosis (TUNEL assay). In addition, there was a significant increase of sub-G1 phase arrest in the cell cycle after sesamin treatment. Furthermore, sesamin increased the expression of apoptotic markers of Bax, caspase-3, and cell cycle control proteins, p53 and checkpoint kinase 2. Taken together, these results suggested that sesamin might be used as a dietary supplement f or prevention of breast cancer by modulating apoptotic signal pathways and inhibiting tumor cell growth.

• Journal of Acupuncture Research
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• v.20 no.3
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• pp.63-74
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• 2003

목적 : 한의학에서 관절염이나 진통치료에 사용되어 왔던 봉독약침액이 인간 골육종 세포주인 MG-63 세포에서 항종양효과가 있는지 연구하고자 한다. 특히 본 실험에서는 이러한 봉독의 종양발생 억제작용이 세포사멸과 관련이 있는지, 그리고 프로스타글란딘 합성 효소인 cyclooxygenase(COX)-2의 억제와 관련이 있는지를 연구하고자 한다. 방법 : 인간 골육종 세포주에서 세포사멸의 변화를 관찰하기 위해서 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium brimide(MTT) assay, 4,6-diamidino-2-phenylindole (DAPI), DNA fragmentation assay 및 reverse transcription-polymerase chain reaction(RT-PCR) 방법을 이용하였다. 결과 : 세포독성 검사에서 봉독은 MG-63 세포에서 농도-의존적으로 세포독성을 나타내었다. 이러한 봉독의 세포독성이 세포사멸로 인한 것인지를 여러 가지 형태로 검사한 결과 봉독에 의한 세포독성은 TUNEL 검사와 DAPI 염색시 세포사멸의 특징적인 소견들을 나타내었고, flow cytometric 분석에서도 세포사멸을 의미하는 세포주기의 변화들을 나타내었다. 봉독이 COX-2의 발현에 미치는 영향을 RT-PCR로 실험한 결과 봉독은 COX-2 mRNA의 발현을 선택적으로 억제하였다. 결론 : 본 실험의 결과 봉독은 COX-2 mRNA의 발현을 억제함으로써 골육종 세포에서 세포사멸을 유발하고 그 결과 항종양효과를 나타내는 것으로 보여진다.