Taxol(paclitaxel) is used in chemotherapy against several cancer. Treatment of tumor cell lines with taxol induces apoptosis, but exact mechanism is not sufficiently understood. The purpose of this study was to investigate apoptosis by the inhibitory effect of paclitaxel on the motility properties of human salivary gland adenocarcinoma cell lines. Paclitaxel inhibited cell motility induced by soluble and immobilized attractant. It suggested that paclitaxel would be a potent inhibitor of salivary gland adenocarcinoma cell motility independent of its cytotoxic and apoptotic activity.

This study investigated the developmental ability and gene expression of somatic cell nuclear transfer embryos using ear skin fibroblast cells derived from miniature pig. When miniature pig (m) and landrace pig (p) were used as donor cells, there were no differences in cleavage (79.2 vs. 78.2%) and blastocyst rates (27.4 vs. 29.7%). However, mNT blastocysts showed significantly higher apoptosis rate than that of pNT blastocysts (6.1 vs. 1.7%) (p<0.05). The number of nuclei in pNT blastosysts was significantly higher than that of mNT (35.8 vs. 29.3) (p<0.05). Blastocysts were analyzed using Realtime RT-PCR to determine the expression of Bax-, Bcl-xl, H19, IGF2, IGF2r and Xist. Bax- was higher in mNT blastocyst than pNT blastocyst (p<0.05). There was no difference in Bcl-xl between two NT groups. Bax-/Bcl-xl was, however, significantly higher in mNT blastocyst compared to pNT. The expression of imprinting genes were aberrant in blastocysts derived from NT compared to in vivo blastocysts. H19 and IGF2r were significantly lower in mNT blastocysts (p<0.05). The expression of IGF2 and Xist was similar in two NT groups. However, imprinting genes were expressed aberrantly in mNT compared to pNT blastocysts. The present results suggest that the NT between donor cells derived from miniature pig and recipient oocytes derived from crossbred pig might affect reprogramming of donor cell, resulting in high apoptosis and aberrant expression patterns of imprinting genes.

Cordyceps militarisis well known as a traditional herbal ingredient, which has been used for patients suffering from cancer in oriental medicine. In this study we have investigated the biochemical mechanisms of anti-proliferative effects by C. militarisextract(CME) in human breast cancer MDA-MB-231 cells. It was found that CME treatment induced chromatin condensation, mitochondrial energization, annexin V staining and sub-G1 phase DNAcontent. These indicators of apoptosis correlate with the mitochondrial dependent pathway, which results in the activation of caspase-3 activity. Both the cytotoxic effect by CME treatment were significantly inhibited by z-DEVD-fmk, a caspase-3 inhibitor,demonstrating the important role of caspase-3 in the observed cytotoxic effect. Cotreatment of CME and LY294002, resulted in significantly induction of apoptosis. These results indicate that caspase-3 is a key regulator of apoptosis in response to CME in human breast cancer MDAMB- 231 through downregulation of Akt, and that the C. militaris extract may therefore have therapeutic potential against human breast cancer.

Agaricus blazei is well known as a traditional medicinal mushroom and it has been shown to exhibit immunostimulatory and anti-cancer activity. However, the cellular and molecular mechanism of apoptosis of cancer cells is poorly understood. In this study, we have investigated whether A. blazei extract (ABE) exerts anti-proliferative and apoptotic effects on human leukemia THP-1 cells. It was found that ABE induced a time- and dose-dependent increase in leukemia cells apoptosis through caspase-3 activation and PARP cleavage. Activation of caspase- 9 induced by ABE suggested that ABE-induced signaling was mediated through a mitochondrial death pathway. In addition, we observed an elevation of ROS and a consequent loss of mitochondrial membrane potential, further suggesting that ABE-induced death signaling was mediated through a mitochondrial oxygen stress pathway. The antioxidant Nacetylcysteine, however, opposed ABE-mediated mitochondrial dysfunction, caspase activation, and apoptosis, supporting the role of ROS in the apoptotic process. We conclude that ABE induces apoptosisin human leukemia cells through a reactive oxygen species and caspase-dependent mitochondrial pathway.

In this study, the apoptotic effects of the actin disruption agent, latrunculin B(LB) have been investigated on p53 deficient chronic myeloid leukemia cell line K562. A dose-dependent decrease in K562 cell proliferation was observed after LB treatment with maximum decrease in cell proliferation being at 1.5μM where the percent inhibition was 66.53%. F-actin stained with TRITC-phalloidin was shown as a peripheral ring or appeared diffusely distributed throughout the cytoplasm in untreated cells, this actin ring was decreased following LB treatment, and even large focal actin aggregates were formed. Treatment of K562 with LB(1.5μM) generated ROS substantially. LB activated expression in a dose-dependent manner. Therefore it can be concluded that LB, depolymerising agent of actin, induces apoptosis by producing ROS and up-regulating NF-kB and COX-2 activation.

Inhibition of proteasome activity may reduce many types of cancer, so it's pathway is effective in cancer as well as in clinical fields. Here the author has carried out experiment targeting on the elevation of apoptosis in oral cancer cells by combination of proteasome inhibitor, lactacystin, and DNA replication inhibitor, etoposide. The growth of KB cells was measured by MTT methods and apoptosis was analyzed by DNA fragmentation and Hochest nucleus staining. The proteasome activity was analyzed by fluorescent tagged peptide and cellular protein expression was detected by Western hybridization. Though lactacystin and etoposide inhibited KB cell growth alone, but low combined doses inhibited cell growth more strongly and induced apoptosis. The proteasome activity was also seriously inhibited by the combination of both chemicals. Tumor suppressor proteins and apoptosis inducing proteins were highly increased under the combination of both chemicals. From above studies we can conclude that proteasome inhibitors may be used for the treatment of oral cancer and proteasome inhibitors with DNA replication inhibitors may be effective in clinical trials of oral cancer.

The prominent side effect of cyclosporin A, immunosuppressive agent, in oral tissues is gingival overgrowth(GO). It is characterized by the gingival enlargement with epithelial thickening, a large number of proliferating fibroblasts and overproduction of ECM components. Fibroblast accumulation in Cs A induced GO is caused by the Inhibition of apoptosis. But CsA effect on normal human oral keratinocyte(NHOK) remains unclear. Transglutaminase 2(TGase 2) which is expressed and activated in tissue where epithelial cells undergo apoptosis has been used as a marker for apoptotic cells. The purpose of this study were to study the effect of CsA on the proliferation and apoptosis of cultured NHOK by TGase 2 expression. After primary cultured NHOK was treated by 0.1, 1.0 and 10ug/ml Cs A, growth curve, MTT assay for succinyl dehydrogenase activity and RT-PCR for TGase 2 mRNA expression were done. The obtained results were as follows. MTT assay showed about 65% cell viability at 1.0μg/ml and 40% at 10μg/ml CsA. Growth curve showed normal S curve on control & DMSO, while decreased growth rate after 3 days of higher CsA tx. TGase 2 mRNA expression of cultured NHOK was the highest at 10μg/ml Cs A. TEM showed chromosome margination, and vacuole formation and clustered mitochodria in cytoplasm of cultured NHOK after CsA tx. It suggested that higher CsA might induce apoptosis of NHOK correlated with increased TGase 2 mRNA expression

Oral squamous cell carcinoma (OSCC) is the most common malignancy and is a major cause of worldwide cancer mortality. The proto-oncogene c-myb plays an important role in regulation of cell growth and differentiation, and it is expressed at high levels in hematopoietic cells and many other types of cancers. However, the function of c-myb is not well known in OSCC. The present study aimed to reveal the function of c-myb and to test the alternation of cell growth and signaling by c-myb in OSCC. In this study, c-myb and dominant-negatibe myb(DNmyb) were expressed in an adenovirus-mediated gene delivery system to KB cells. The over-expressed c-myb brought increased cellular proliferation compared with control cells. However, DN-myb infected KB cells showed significant reduction of cell growth and enhanced induction of apoptosis to activate PARP and caspase 9. c-myb induced increase of IGF-I, -II and IGF-IR expressions while DN-myb down-regulated these expression. Activation of ERK and Akt/PKB pathway was shown only in c-myb transduced cells. These findings suggest that the role of c-myb in cell growth of oral cancer cells is partially mediated through the modulation of IGFs, ERK and Akt/PKB. From this results, DN-myb is strongly recommended as a curable gene for the treatment of c-myb dependent malignancies such as OSCC.

This study was conducted to examine the mRNA expression of apoptosis-related and imprinted genes and methylation pattern of the differentially methylated region (DMR) of H19 gene in day 35 of SCNT pig fetuses. The day 35 of natural mating (control) or cloned (clone) pig fetuses were recovered from uterus. Endometrium from dam and liver from fetus were obtained, respectively. mRNA expression was evaluated by real-time PCR and methylation pattern was analyzed by bisulfite sequencing method. The Bcl-2 mRNA expression in clone was significantly lower than that of control (p<0.05). The mRNA expression of H19 gene in both endometrium and liver was significantly higher in clone than that of control, respectively (p<0.05). The level of IGF-2 mRNA in liver of clone was significantly lower than that of control (p<0.05), whereas the mRNA expression of IGF2-R gene in liver of clone was significantly higher than that of control (p<0.05). The DMR of H19 was lower methylation pattern in clone than that of control. These results suggest that the aberrant mRNA expression of apoptosis-related and imprinted genes and the lower DMR methylation pattern of imprinted gene may be closely related to the inadequate fetal development of cloned fetus.

Cordyceps militaris is well known as a traditional herbal ingredient, which has been used for patients suffering from cancer in oriental medicine. In this study we have investigated the biochemical mechanisms of anti-proliferative effects by C. militaris extract(CBE) in human breast cancer MDA-MB-231 cells. It was found that CBE treatment induced chromatin condensation, mitochondrial energization, annexin V staining and sub-G1 phase DNA content. These indicators of apoptosis correlate with the mitochondrial dependent pathway, which results in the activation of caspase-3 activity. Both the cytotoxic effect by CBE treatment were significantly inhibited by z-DEVD-fmk, a caspase-3 inhibitor, demonstrating the important role of caspase-3 in the observed cytotoxic effect. Co-treatment of CBE and LY294002, resulted in significantly induction of apoptosis. These results indicate that caspase-3 is a key regulator of apoptosis in response to CBE in human breast cancer MDA-MB-231 through down regulation of Akt, and that the C. militaris extract may therefore have therapeutic potential against human breast cancer.

Glycyrrhiza ura lensis (Leguminosae) has long been known as an ant i-i nflammatory agent for gastric 띠cers ， arthri t is, and rheumaLism. 1'he f1avonoid glycyr ol isolated from Glycyrrhiza uralensis dramatically inhibits phorbol ester (PMA)-induced NF-kB-dependent transcriptional activity, as determined by luciferase activity in HEK293T cells To in vestigate the global gene expression profiling by glycyrol, we performed high-density oligonucleotide microarrays, Our microarray analysis showed that gl ycY I 이 inhi bited phorbol ester-induced NF-kB-dependent transcriptional activity in inflarnmatory-related gene expression, RT-PCR analysis based on microarry data showed that NF-kB-dependent genes such as CCL2‘ CCL7. CD44, and HSPB8 in addition to NF-kB itself were significantly down-regulated by glycyrol Treatment with glycyrol inhibited ]-kB degradation induced by PMA, suggesting that glycyrol has a potential anti-in flammatory activity by modulating NF-kB-dependent pathways. Also, the microarray data showed that glycyr이 appears to induce gene expression related to p53-dependent apoptosis t hrough ENDO G instead of CAD/DFF and AlF/PDCD8 as down stream apoptosis factor in HEK293T tumor cell s and to induce an added function as oncogenes in connection with apoptoslS

Treatment of oral cancers with chemotherapeutic agents become evaluated as an effective method to reduce cancer cell proliferation. Anti-proliferative and anti-oral cancer activities of momordin I on oral cancer cells were evaluated in this study. Momordin I was originally purified from a natural product, Ampelopsis radix and showed the antiproliferative activity against oral carcinoma, KB cells. Obtained value was approximately 104μM/mℓ. Time-and dose-dependent chromosomal DNA fragmentations were observed in momordin I-treated KB cells. Flow cytometry analysis showed time-dependent apoptotic cell appearance after treatment of momordin I. Approximately 18.6% apoptotic cells were observed at 72 hours after of momordin I treatment. These observation were consistent with the results obtained in DNA fragmentation analysis. These data suggest that momordin I has anti-proliferative effect and induces cell death in KB cells through apoptosis.

Disruption of cell - matrix attachment results in a loss of prosurvival signals and culminates in programmed cell death, referred to as anoikis , Apoptosis signal- regulating kinase 1(ASKl)/MKK5 is a ubiquitously expressed enzyme that acti vates c-Jun N-terminal kinase/stress-activated protein kinase JNK/SAPK and p38 pathways by direct site specific Ser/Thr phosphoryl ation of their respective MKKs-MKK4/MKK7 for JNK and MKK3/MKK6 for p38 kinases, The kinase activity of ASKl is stimulated by a variety of death signals, including TNF, Fas ligation, reactive oxygen species, and antineoplastic agents , The aim of this study was to investigate the relative importance of ASKl in anOlkls 1n the present study cells which lost their adhesion showed higher rate of cell death in compared to cells which maintained anchorage. 1nterestingly the res ult showed that suspended cells expressing ASK1 were more susceptible to anoikis than suspended cells having no ASK1 1n addition, cellu lar attachment seems to have significant effect on ASKl activity and p38 MAPK protein rather than serum stimulation