Cruciferous vegetables including diindolylmethane (DIM) have been shown to have anticancer activity. Especially, DIM-pPhBr and DIM-pPhF used in this study was reported to have more effective and less toxic effects than DIM. However, there is no report presenting their anti-tumorigenic activity in oral cancer. In the present study, we examined the effects of DIM-pPhBr and DIM-pPhF on the cell proliferation and apoptosis in KB human oral cancer cells. DIM-pPhBr and DIM-pPhF decreased cell proliferation and induced apoptosis evidenced by western blot analysis, DAPI staining and sub-G1 population. This provides the first evidence that DIM-pPhBr and DIM-pPhF originating from cruciferous vegetables induce apoptotic cell death in human oral cancer cells to inhibit cancer cell proliferation.

본 연구에서는 효모에서 과 발현하는 Bax inhibior와 관련된 유전자를 동정하여 특성화 하였다. Yeast functional screening이라는 방법을 이용하여, 일반적은 환경에서 재배된 벼의 cDNA를 QX95001에 형질전 환하여 SD-galactose-Leu--Ura-배지에서 생성된 8개의 클론을 선발하였다. 그 중 AtBI-1과 같은 domain이 있는 D2-234를 포함하여 5개의 클론을 선발하였다. D2-243는 741bp의 염기서열과 247개의 아미노산으로 구성되었고 5 membrane-spanning 단편으로 되어 있음을 확인하였다. D2-234는 SD-galactose--배지에서 세포성장이 왕성하였다. 본 실험에서 얻어진 결과는 벼 식물에서 나타나는 세포예정사와 관련된 단백질을 선발하는데 유용하게 이용될 것으로 생각된다.

(‐)‐Epigallocatechin 3‐gallate (EGCG) is a potent antioxidant polyphenol in green tea that acts as an anticancer agent via both direct and indirect pathways. Although the relationship between EGCG’s anticancer effects and its antioxidant activity is not fully understood, it is known that EGCG stimulates production of reactive oxygen species (ROS), which induce oxidative stress leading to cell death. In IM9 multiple myeloma cells, EGCG acted in a dose‐ and time‐dependent manner to induce apoptotic cell death. Among the antioxidant enzymes expressed in IM9 cells, levels of peroxiredoxin (Prdx) V were selectively and significantly reduced by EGCG. Moreover, the ROS scavenger NAC completely inhibited EGCG‐induced apoptosis and PrdxV reduction, while overexpression of PrdxV, but not a PrdxVC48S mutant, protected IM9 cells from EGCG‐induced apoptosis. EGCG‐induced reductions in cell viability and PrdxV levels were also observed in primary CD138+ multiple myeloma cells from patients. These results suggest that PrdxV is a key target via which EGCG mediates its anticancer effects.

In this study, we analyzed expression patterns of apoptotic and autophagic gene products in culture follicular cells of normal and miniature pigs to assess the effect of hormones on the choice for programmed cell death. Autophagic activity progressively increased from control cultures to luteinizing hormone (LH)-treated cultures of follicular cells of normal pigs, but decreased from the LH to follicle stimulating hormone (FHS) +LH-treated cultures. Expression of Casp-3 protein in follicular cells was highest in LHtreated cultures, but the activity of Casp-3 decreased in the control, FSH-treated, and FSH+LH-treated cultures. The activity of the apoptosis protein was highly expressed in the control, LH-treated, and FSH+LH-treated follicular cells of miniature pigs, but autophagy- associated proteins were expressed at low levels in all treatments groups of the miniature pig. The expression of autophagy and apoptosis proteins appeared similar in control and rapamycin-treated cells. In addition, stimulation with FSH triggered the activation of autophagy in the follicular cells of normal pigs, but induced apoptosis in the follicular cells of miniature pigs. A similar effect was obtained when LH was applied. These results suggest that the autophagy process and FSH stimulation is more effective for stable and innovative follicular cell development.

Betulinic acid (BA), a naturally occurring triterpene found in the bark of the white birch tree, has been investigated to induce apoptosis in various cancer cells and animal models. However, there is no report of the chemopreventive effect of BA in cervical cancer cells. Using KB human cervical cancer cells as a model, we currently show that BA decreases cell viability and induces apoptotic cell death. The mechanism of the BA-induced anti-growth response in KB cells is due to the down-regulation of specificity protein 1 (Sp1) and its downstream targets, myeloid cell leukemia-1(Mcl-1) and survivin. Thus, BA acts as a novel chemopreventive agent through the regulation of Sp1that is highly expressed in tumors.

Chios gum mastic (CGM) is produced from Pistiacia lentiscus L var chia, which grows only on Chios Island in Greece. CGM is a kind of resin extracted from the stem and leaves, has been used for many centuries in many Mediterranean countries as a dietary supplement and folk medicine for stomach and duodenal ulcers. CGM is known to induce cell cycle arrest and apoptosis in some cancer cells. This study was undertaken to investigate the alteration of the cell cycle and induction of apoptosis following CGM treatment of HL-60 cells. The viability of the HL-60 cells was assessed using the MTT assay. Hoechst staining and DNA electrophoresis were employed to detect HL-60 cells undergoing apoptosis. Western blotting, immunocytochemistry, confocal microscopy, FACScan flow cytometry, MMP activity and proteasome activity analyses were also employed. CGM treatment of HL-60 cells was found to result in a dose- and time-dependent decrease in cell viability and apoptotic cell death. Tested HL-60 cells showed a variety of apoptotic manifestations and induced the downregulation of G1 cell cycle-related proteins. Taken collectively, our present findings demonstrate that CGM strongly induces G1 cell cycle arrest via the modulation of cell cycle-related proteins, and also apoptosis via proteasome, mitochondrial and caspase cascades in HL-60 cells. Hence, we provide evidence that a natural product, CGM could be considered as a novel therapeutic for human leukemia.

The majority of strokes are caused by ischemia and result in brain tissue damage, leading to problems of the central nervous system including hemiparesis, dysfunction of language and consciousness, and dysfunction of perception. The purpose of this study was to investigate the effects of Poly(ADP-ribose) polymerase(PARP) on necrosis in neuronal cells that have undergone needle electrode electrical stimulation(NEES) prior to induction of ischemia. Ischemia was induced in male SD rats(body weight 300g) by occlusion of the common carotid artery for 5 min, after which the blood was reperfused. After induction of brain ischemia, NEES was applied to Zusanli(ST 36), at 12, 24 and 48 hours. Protein expression was investigated using immuno-reactive cells, which react to PARP antibodies in cerebral nerve cells, and Western blotting. The results were as follows: In the cerebral cortex, the number of PARP reactive cells after 24 hours significantly decreased(p<.05) in the NEES group compared to the GI group. PARP expression after 24 hours significantly decreased(p<.05) in the NEES group compared to the GI group. As a result, NEES showed the greatest effect on necrosis- related PARP immuno-reactive cells 24 hours after ischemia, indicating necrosis inhibition, blocking of neural cell death, and protection of neural cells. Based on the results of this study, NEES can be an effective method of treating dysfunction and improving function of neuronal cells in brain damage caused by ischemia.

The use of bacteria in the treatment of cancer has a long and interesting history. The use of live bacteria in this way however has a number of potential problems including toxicity. Purified low molecular weight bacterial proteins have therefore been tested as anticancer agents to avoid such complications. Oral cancer is a widely occurring disease around the world and these lesions are typically very resistant to anticancer agents. In our present study we investigated the effects of purified recombinant azurin from Pseudomonas (P.) aeruginosa against YD-9 (p53-positive) human oral squamous carcinoma cells. Azurin showed cytotoxic effects against these cells in a dose dependent manner. The cell death accompanied by this treatment was found to be characterized by chromatin condensation and apoptotic bodies. Azurin treatment was further found to increase the expression of p53 The stabilization of p53 and induction of apoptosis in YD-9 cells by azurin suggests that it has potentially very strong anticancer properties in oral squamous carcinoma.

Melatonin (N-acetyl-5-methoxytryptamine) is the major neurohormone secreted during the night by the vertebrate pineal gland. The circadian pattern of pineal melatonin secretion is related to the biological clock within the suprachiasmatic nucleus (SCN) of the hypothalamus in mammals. The SCN coordinates the body's rhythms to the environmental light-dark cycle in response to light perceived by the retina, which acts mainly on retinal ganglion cells that contain the photopigment melanopsin. Calbindin-D9k (CaBP-9k) is a member of the S100 family of intracellular calcium- binding proteins, and in this review, we discuss the involvement of melatonin and CaBP-9k with respect to calcium homeostasis and apoptotic cell death. In future studies, we hope to provide important information on the roles played by CaBP-9k in cell signal transduction, cell proliferation, and homeostasis in vivo and in vitro.

Adherent cells, such as those found in epithelial tissues, must be physically associated with extracellular matrix (ECM)components to survive. Though stimulation by growth factors is an essential factor in cell survival, normal cells also requires cell adhesion to ECM proteins. The cessation of these anchorage-mediated signals seems to be a common mechanism to physiolog ically t erminate t he l ife cycle of t hese c ells b y apoptosis. This form o f cell death has been termed anoikis.In cancer, resistance to anoikis of cancer cell is important in invasion and metastasis. The present study investigated the intracellular mechanism involved in anoikis, especially in cells treated with epigallocatechin- 3-gallate(EGCG). To induce anoikis, cell culture plates were coated with 10 ㎍/ml poly-HEMA. A549 lung adenocarcinoma cells were grown in RPMI 1640 medium with/without 10% fetal bovine serum, and then cells were replated on cell culture dishes coated with poly-HEMA in the presence or absence of serum. On the other hand, EGFR inhibitor, PI3K inhibitor, and EGCG were treated to the anoikis status cells, in order to evaluate the factors of anoikis. The result revealed that growth factors or loss of adhesion can increase phosphorylate Akt. In addition, lack of cell adhesion fails to activate pro-apoptotic factors directly. Activity of Erk kinase depends on not only EGFR signaling but also cell adhesion. Akt activation is mainly affected by EGCG whereas Raf-1 activation is controlled by the presence of cell contact. In addition, EGCG increased the level of NFkB, whereas phophroylated PTEN and total PTEN were not different. In this report,increase of NFkB was correlated with Akt phosphorylation, suggesting that EGCG can protect cells from detachment–induced cell death through Akt activation and subsequent NFkB

Green tea, derived from the plant Camellia sinensis, is one of the most common beverages consumed worldwide. Epigallocatechin-3-gallate (EGCG) is the most abundant and bioactive polyphenolic constituent in green tea. Understanding how intracellular signaling pathways respond to EGCG may provide a clue to the difference of cell responses and basis for usefulness of EGCG as a chemopreventive and/or chemotherapeutic agent. In the present study, we tried to check whether EGCG could be a useful agent in chemotherapeutic treatment of oral squamous carcinoma. Furthermore, we investigated which signaling pathway is involved in biologic activities of EGCG. EGCG induced the cell death of oral squamous carcinoma cells. Furthermore, it increased phosphorylation of Akt in serum-strarved oral squamous carcinoma cells. But, initial increase of Akt activation did not affect cell survival. Activities of Raf-1 and Erk showed inconsistent response to EGCG treatment, but Erk phosphorylation is consistent with Raf-1 activity in YD 10B cells. These changes of Raf-1 and Erk activity in EGCG treated cells were different depending on cell line type. Supposedly, the difference of cell component may affect the Raf-1 and Erk reactivity to EGCG treatment. Akt activation by EGCG is independent on activities of PDK1 and PTEN, and expression of bax and bcl-2 proteins were not changed by EGCG treatment. Therefore, EGCG treatment did not induce the apoptosis of YD 10B cell. On the other hand, vascular adhesion molecule (VCAM) was decreased by EGCG treatment, so it is possible that decrease of VCAM can play certain role in survival and/or cell death in EGCG treated cells

The Maillard Reaction Products (MRPs) such as Glucose-tyrosine (Glu-Tyr) and Xylose-arginine (Xyl-Arg) have antioxidant, antimutagenic, and antibacterial effects. However, to date, still little is known about the other biological effects of the MRPs. In this study, we investigated whether the fructose-tyrosine MRP, 2,4-bis(p-hydroxyphenyl)-2-butenal (Fru-Tyr), could modulate cell cycle progression and NF-κB activity, and thereby induce apoptotic cell death of colon cancer cells. Treatment with different concentrations (10-40 μg/ ml) of Fru-Tyr for 24 h inhibited colon cancer cell (SW620 and HCT116) growth followed by induction of G₂/M phase cell cycle arrest and apoptosis in a dose-dependent manner. We also found that Fru-Tyr suppressed tumor necrosis factor-alpha (TNF-α)-induced NF-κB transcriptional activity. Moreover, Fru-Tyr induced the expression of apoptotic gene, cleaved caspse-3. These results suggest that Fru-Tyr inhibited colon cancer cell growth through induction of G₂/M phase cell cycle arrest and apoptotic cell death by modulating of NF-κB.

SET (Suppressor of variegation, Enhancer of zeste, and the Trithorax) domain-containing proteins are known to have methyltransferase activity at lysine residues of histone proteins. In this study, we identified a novel SET domain-containing protein from mouse and named Kodo7. Indeed, Kodo7 has methyltransferase activity at K9 residue of the H3 protein as demonstrated by a histone methyl-transferse activity assay using GST-tagged Kodo7. Confocal microscopy showed that Kodo7 is co-localized with histones in the nucleus. Interestingly, ectopic expression of Kodo7 by transient transfection induced cell death and treatment of the transfectants with a caspase-3 inhibitor, Ac-DEVD-AFC decreased Kodo7-induced apoptosis. These results suggest that Kodo7 induces apoptotic cell death through increased methylation of histones leading to transcriptional repression.

Concerns remain regarding the biocompatibility and adverse effects of dental casting alloys. The aim of this study was to understand the cytopathogenic effect of metal ions, which might be released from dental alloys, on oral squamous carcinoma(OSC) cells. The cellular morphology, viability, the type of cell death and molecular change in response to metal ion salt solutions including aluminum(Al), cobalt(Co), copper(Cu) and nickel(Ni) were examined. The values for the metal ions with the exception of AI were estimated to be between 400 and 600μM. The cells treated with the metal ions showed apoptotic change with the exception of Al ions. Metal ion-induced apoptosis was further confirmed using flow cytometric analysis. This study showed that the cytotoxicity and the mode of cell death by metal ions clearly depend on the cell type, the type of metal ion and the duration of exposure. The protein level of Rb, a tumor suppressor that affects apoptosis para-doxically, was higher in the cells treated with Co, Cu and Ni. It is believed that apoptosis and cell damage in the OSC cells treated with Co, Cu or Ni can be evoked by the regulation of Rb.

Nitric oxide (NO) has been known to inf1uence cell fate through apoptotic or necrotic cell death. Here, we investigated the role of nitric oxide on the growth and viability of immortalized human salivary gland (HSG) cells 띠 vitro. Treatrnent of HSG with a NO donor, S-nitroso-N-acetyl-DL-penκi1lamine (SNAP), significantly diminished the growth rate of HSG in a concentration dependent manner. However, this retardation of cell비ar growth rate was not corresponded to the apoptotic cell death of HSG cells, because there were no characteristic apopto디c features such as condensation of nuclear chromatin, nuclear fragmentation, and the apoptotic peak of propidium iodide (PI)-stained nuclei by flow cytome띠. 까ùs implies that HSG cells are resistant to NO-mediated 다π。to:잉city. 1n SNAP treated HSG cells, cell cycle analysis revealed that the number of G2/M phase increased markedly, according to while the percentage of cells in GO/Gl and S phases was not significantly affected. Otherwise, high concentrations of SNAP increased both P1 and annexin V positive cells. 1nterestingly, preincubation of HSG cells with iron chelator, deferoxamine (DFO), significantly diminished NO cytotoxicity more than when HSG cells are only incubated with SNAP which su잃.ests the role of iron homeostasis in NO-mediated cell death of HSG cells. 1n addi디。n ， treatrnent of HSG cells with SNAP specifically cleaved iron regulatory protein-2 (IRP2) while not affecting 1RP1. Collectively, the mπent results s멍gest that NO has a potential to control HSG cell growth through cell cycle arresting at G2/M phase. 1n addi디on ， intracellular iron homeostasis nùght play an important role in regulating cell survival of HSG cells