• Asian Pacific Journal of Cancer Prevention
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• v.13 no.7
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• pp.3103-3107
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• 2012

Osteosarcoma, the most common primary mesenchymal malignant tumor, usually has bad prognosis in man, with cancer stem-like cells (CSCs) considered to play a critical role in tumorigenesis and drug-resistance. It is known that phosphatidylinositol 3-kinase (PI3K) is involved in regulation of tumor cell fates, such as proliferation, cell cycling, survival and apoptosis. Whether and how PI3K and inhibitors might cooperate in human osteosarcoma CSCs is still unknown. We therefore evaluated the effects of LY294002, a PI3K inhibitor, on the cell cycle and apoptosis of osteosarcoma CSCs in vitro. LY294002 prevented phosphorylation of protein kinase B (PKB/Akt) by inhibition of PI3K phosphorylation activity, thereby inducing G0/G1 cell cycle arrest and apoptosis in osteosarcoma CSCs. Further studies also demonstrated that apoptosis induction by LY294002 is accompanied by activation of caspase-9, caspase-3 and PARP, which are involved in the mitochondrial apoptosis pathway. Therefore, our results indicate PI3K inhibitors may represent a potential strategy for managing human osteosarcoma via affecting CSCs.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.7
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• pp.3471-3476
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• 2012

Background and Objective: Histone deacetylase (HDAC) inhibitors represent a promising class of potential anticancer agents for treatment of human malignancies. In this study, we investigated the effect of trichostatin A (TSA), one such HDAC inhibitor, in combination with docetaxel (TXT), a cytotoxic chemotherapy agent or erlotinib, a novel molecular target therapy drug, on lung cancer A549 cells. Methods: A549 cells were treated with TXT, erlotinib alone or in combination with TSA, respectively. Cell viability, apoptosis, and cell cycle distribution were evaluated using MTT (3- (4, 5-dimethylthiazol-2-yl) -2, 5-diphenyltetrazolium bromide) assay, Hochst33258 staining and flow cytometry. Moreover, immunofluorescent staining and Western blot analysis were employed to examine alterations of ${\alpha}$-tubulin, heat shock protein 90 (hsp90), epidermal growth factor receptor (EGFR), and caspase-3 in response to the different exogenous stimuli. Results: Compared with single-agent treatment, co-treatment of A549 cells with TSA/TXT or TSA/erlotinib synergistically inhibited cell proliferation, induced apoptosis, and caused cell cycle delay at the $G_2/M$ transition. Treatment with TSA/TXT or TSA/erlotinib led to a significant increase of cleaved caspase-3 expression, also resulting in elevated acetylation of ${\alpha}$-tubulin or hsp90 and decreased expression of EGFR, which was negatively associated with the level of acetylated hsp90. Conclusions: Synergistic anti-tumor effects are observed between TXT or erlotinib and TSA on lung cancer cells. Such combinations may provide a more effective strategy for treating human lung cancer.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.4
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• pp.1947-1959
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• 2016

Background: Colorectal cancer (CRC) is a major cause of morbidity and mortality, being the second most common type of cancer worldwide in both men and women. It accounts yearly for approximately 9% of all new cases of cancers. Furthermore, the current chemotherapeutic regimens seem unsatisfactory, so that exploration of novel therapeutic modalities is needed. The present study was undertaken to investigate the inhibitory effects of a crude alkaloid extract (CAERS) of a medicinal herb, Rhazya stricta, on proliferation of CRC HCT116 cells and to elucidate mechanisms of action. To achieve these aims, we utilized MTT, comet, DNA laddering and gene reporter assays, along with Western blot and RT-PCR analyses. Results: We found that CAERS inhibited cell proliferation and induced apoptotic cell death in HCT116 cells. Hallmarks of morphological and biochemical signs of apoptosis were clearly evident. CAERS down-regulated DNA-binding and transcriptional activities of NF-${\kappa}B$ and AP-1 proteins, while up-regulating expression of the Nrf-2 protein. It also down-regulated expression levels of the ERK MAPK, Bcl-2, cyclin D1, CDK-4, survivin and VEGF and up-regulated levels of Bax, caspase-3/7 and -9, p53, p21, Nrf-2. Markedly, it promoted mRNA expression levels of cytoprotective genes including the hemeoxygenase-1, NAD(P)H quinine oxidoreductase 1 and UDP-glucuronyltransferase. Conclusions: These findings indicate that CAERS exerts antiproliferative action on CRC cells through induction of apoptotic mechanisms, and suggest CAERS could be a promising agent for studying and developing novel chemotherapeutic agents aimed at novel molecular targets for the treatment of CRC.

• Toxicological Research
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• v.35 no.1
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• pp.83-91
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• 2019

Nanoparticles (NPs) have been recognized as both useful tools and potentially toxic materials in various industrial and medicinal fields. Previously, we found that zinc oxide (ZnO) NPs that are neurotoxic to human dopaminergic neuroblastoma SH-SY5Y cells are mediated by lipoxygenase (LOX), not cyclooxygenase-2 (COX-2). Here, we examined whether human bone marrow-derived mesenchymal stem cells (MSCs), which are different from neuroblastoma cells, might exhibit COX-2- and/or LOX-dependent cytotoxicity of ZnO NPs. Additionally, changes in annexin V expression, caspase-3/7 activity, and mitochondrial membrane potential (MMP) induced by ZnO NPs and ZnO were compared at 12 hr and 24 hr after exposure using flow cytometry. Cytotoxicity was measured based on lactate dehydrogenase activity and confirmed by trypan blue staining. Rescue studies were executed using zinc or iron chelators. ZnO NPs and ZnO showed similar dose-dependent and significant cytotoxic effects at concentrations ${\geq}15{\mu}g/mL$, in accordance with annexin V expression, caspase-3/7 activity, and MMP results. Human MSCs exhibited both COX-2 and LOX-mediated cytotoxicity after exposure to ZnO NPs, which was different from human neuroblastoma cells. Zinc and iron chelators significantly attenuated ZnO NPs-induced toxicity. Conclusively, these results suggest that ZnO NPs exhibit both COX-2- and LOX-mediated apoptosis by the participation of mitochondrial dysfunction in human MSC cultures.

• Clinical and Experimental Reproductive Medicine
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• v.51 no.3
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• pp.192-204
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• 2024

Objective: Cisplatin (CP) is a widely used chemotherapeutic agent, but its severe side effects impact testicular function. We investigated the potential protective effects of bilberry extract against CP-induced testicular toxicity. Methods: Forty adult male albino rats were divided into four groups. Control animals received a single oral dose of 0.9% saline. Bilberry-treated rats received oral bilberry extract (200 mg/kg body weight [BW] dissolved in 1 mL of saline) daily for 10 consecutive days. CP-treated animals were administered a single intraperitoneal dose (7.5 mg/kg BW). Finally, a bilberry+CP group received oral bilberry extract (200 mg/kg BW) daily for 10 consecutive days, with one intraperitoneal dose of CP (7.5 mg/kg BW) on day 2. We assessed sperm count, motility, viability, and abnormalities, along with testis weight, testis weight-to-BW ratio, antioxidant activity, levels of oxidative stress markers (malondialdehyde [MDA] and hydrogen peroxide [H₂O₂]), sex hormones (follicle-stimulating hormone [FSH], luteinizing hormone [LH], and testosterone), and apoptotic and anti-apoptotic markers, and DNA damage. Testicular tissue underwent histopathological examination. Results: Among CP-treated rats, significantly lower values were observed for testis weight; testis weight-to-BW ratio; levels of FSH, LH, testosterone, superoxide dismutase, catalase, glutathione S-transferase, glutathione, and B-cell lymphoma 2; and sperm count, motility, and proportion of normal sperm. CP administration was associated with higher MDA, H₂O₂, p53, Bax, cytochrome c, caspase 9, and caspase 3 levels, along with elevated tail moment. However, bilberry extract administration significantly improved all altered parameters. Conclusion: Bilberry treatment demonstrated protective effects and reduced CP-induced testicular toxicity via antioxidant activity and cytoprotection.

• Toxicological Research
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• v.21 no.2
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• pp.107-113
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• 2005

[ $BARODON^{(R)}$ ] is a multi-purpose, high functional alkali solution made by mixing and liquid-ionizing silicon, calcium, sodium, borax, organic carbon chemicals and silver. In this study, we have investigated the apoptotic potential and mechanistic insights of $BARODON^{(R)}$ in human gastric cancer cell line (AGS cells). In MTT assay, $BARODON^{(R)}$ reduced cell viability in AGS cells. Morphological features of apoptosis with marked cytoplasmic vacuolation and appearance of apoptotic peaks in flow cytometry were observed in AGS cells with$BARODON^{(R)}$ treatment. In addition, $BARODON^{(R)}$ induced apoptosis of stomach cancer cell is related to bax up-regulation, caspase 7 protease activation and subsequent cleavage of poly (ADP-ribose) polymerase (PARP). These results suggest that BARODON can induce the apoptosis of AGS cells through modulation of bcl-2 family and the activation of intrinsic caspase cascades, indicating that it is potentially useful as a anti-cancer agent.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.7
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• pp.1022-1028
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• 2013

Chaga mushroom (Inonotus obliquus) was considered a functional food with an anti-cancer effect in colon, gastric, and lung cancer. Therefore, this study was conducted in order to elucidate the effect of chaga mushroom extract in brain cancer. Glioblastoma U-87 MG cells were used in investigation of cell survivability, apoptosis, and cell cycle arrest analysis. Treatment with various concentrations of chaga mushroom extract resulted in inhibition of cell proliferation and cell cycle arrest. Although caspase-3 expression was increased over $100{\mu}g/mL$ of chaga mushroom extract treatment, apoptosis factors with Bcl-2, Bax and p53 did not change. In analysis of cell cycle regulatory factors, expression of cyclin D1 and CDK2 decreased in a dose-dependent manner. We have demonstrated the anti-cancer effect of chaga mushroom extract in glioblastoma, which may be mediated by activation of the caspase pathway and induction of cell cycle arrest.

• Korean Journal of Plant Resources
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• v.31 no.2
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• pp.102-108
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• 2018

Rubiae radix is root of Runia akane Nakai, it has been used to hemostasis and blood stasis in Korean and China. This study investigated that anti-oxidant and anti-colorectal cancer effect of ERA (ethanol extract of Rubiae radix) and WRA (water extract of Rubiae radix) using RAW 264.7 (murine macrophage from blood) and HCT-116 cells (human colorectal cancer cell line). ERA contained polyphenol ($45.77{\pm}2.03mg/g$) and flavonoid ($22.82{\pm}1.33mg/g$). $500{\mu}M$ $H_2O_2$-induced ROS generation was diminished by $500{\mu}g/m{\ell}$ ERA treatment in RAW 264.7 cells, but not WRA (125, 250, and $500{\mu}g/m{\ell}$). Moreover, caspase-3 activity and DNA fragmentation increased by $500{\mu}g/m{\ell}$ ERA treatment during apoptotic cell death in HCT-116. Results demonstrated that anti-cancer effect of ERA against human colorectal cancer cells is mediated apoptotic cell death and DNA fragmentation through caspase-3 activation. However, further study is required to what active ingredient of ERA are important for anti-oxidant and anti-colorectal cancer effect in vivo.

• Journal of Life Science
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• v.17 no.7 s.87
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• pp.948-955
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• 2007

Neutrophils play a key role as a first line of defense and are known to acquire the characteristics of dendritic cells (DCs) under the appropriate conditions. The spontaneous apoptosis of neutrophils was delayed by treatment with 4-(2-aminoethyl) benzensulfonylfluoride (AEBSF), a serine protease inhibitor. AEBSF inhibited both caspase-3 and serine protease activities, whereas ZVAD-fmk, a pancaspase inhibitor, inhibited only caspase-3 activity. The life span of neutrophils was prolonged up to 5 days by AEBSF in the presence or absence of granulocyte macrophage colony stimulating factor(CM-CSF). DC surface markers, such as CD80, CD83, and MHC class ll were not expressed on neutrophils treated with AEBSF alone. CM-CSF failed to prolong the survival time of neutrophils up to3 days but increased the expression levels of DC markers on neutrophils in the presence of AEBSF. Expression levels of DC markers were the highest on neutrophils treated with CM-CSF and AEBSF for 3 days. AEBSF and CM-CSF-treated neutrophils stimulated proliferation of T cells in the presence of a superantigen, Staphylococcal enterotoxin B (SEB) but produced $interferon-{\gamma}$ ($IFN{\gamma}$) in the absence of SEB. These results suggest that the inhibition of serine protease activity prolonged the life span of human neutrophils and combined treatment of neukophils with CM-CSF and serine protease inhibitor induced differentiation of neutrophils into DC-like cells.

• Journal of Life Science
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• v.23 no.7
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• pp.926-932
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• 2013

Glutamine and serum are essential for cell survival and proliferation in vitro, yet the signaling pathways that sense glutamine and serum levels in endothelial cells remain uninvestigated. In this study, we examined the effects of glutamine deprivation and serum starvation on the fate of endothelial cells using a human umbilical vein endothelial cell (HUVEC) model. Our data indicated that glutamine deprivation and serum starvation trigger a progressive reduction in cell viability through apoptosis induction in HUVECs as determined by DAPI staining and flow cytometry analysis. Although the apoptotic effects were more predominant in the glutamine deprivation condition, both apoptotic actions were associated with an increase in the Bax/Bcl-2 (or Bcl-xL) ratio, down-regulation of the inhibitor of apoptosis protein (IAP) family proteins, activation of caspase activities, and concomitant degradation of poly (ADP-ribose) polymerases. Moreover, down-regulation of the expression of Bid or up-regulation of truncated Bid (tBid) were observed in cells grown under the same conditions, indicating that glutamine deprivation and serum starvation induce the apoptosis of HUVECs through a signaling cascade involving death-receptor-mediated extrinsic pathways, as well as mitochondria-mediated intrinsic caspase pathways. However, apoptosis was not induced in cells grown in glutamine- and serum-free media when compared with cells exposed to glutamine deprivation or serum starvation alone. Taken together, our data indicate that glutamine deprivation and serum starvation suppress cell viability without apoptosis induction in HUVECs.