• International Journal of Oral Biology
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• v.39 no.1
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• pp.23-33
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• 2014

Several studies have shown that curcumin, which is derived from the rhizomes of turmeric, possesses antimicrobial, antioxidant and anti-inflammatory properties. The antitumor properties of curcumin have also now been demonstrated more recently in different cancers. This study was undertaken to investigate the modulation of cell cycle-related proteins and the mechanisms underlying apoptosis induction by curcumin in the SCC25 human tongue squamous cell carcinoma cell line. Curcumin treatment of the SCC25 cells resulted in a time- and dose-dependent reduction in cell viability and cell growth, and onset of apoptotic cell death. The curcumin-treated SCC25 cells showed several types of apoptotic manifestations, such as nuclear condensation, DNA fragmentation, reduced MMP and proteasome activity, and a decreased DNA content. In addition, the treated SCC25 cells showed a release of cytochrome c into the cytosol, translocation of AIF and DFF40/CAD into the nuclei, a significant shift in the Bax/Bcl-2 ratio, and the activation of caspase-9, caspase-7, caspase-6, caspase-3, PARP, lamin A/C, and DFF45/ICAD. Furthermore, curcumin exposure resulted in a downregulation of G1 cell cycle-related proteins and upregulation of $p27^{KIP1}$. Taken together, our findings demonstrate that curcumin strongly inhibits cell proliferation by modulating the expression of G1 cell cycle-related proteins and inducing apoptosis via proteasomal, mitochondrial, and caspase cascades in SCC25 cells.

• The Korean Journal of Cytopathology
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• v.7 no.1
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• pp.44-50
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• 1996

Small cell carcinoma of the lung is characterized by cells with finely stippled chromatin and scanty cytoplasm as well as a particularly aggressive clinical course and favorable response to the chemotherapy. Recently percutaneous fine needle aspiration (FNA) biopsy has become both widely established and highly respected for the diagnosis of lung cancer. However metastatic small cell carcinoma of lymph node should be cytologically differentiated from the small round cell tumor of particular sites, especially malignant lymphoma, because small ceil carcinoma of classic oat cell type nay simulate small cell non-Hodgkin's lymphoma. We report five cases of metastatic small cell carcinoma of in-termediate cell type diagnosed by FNA of the enlarged lymph nodes of the neck and axilla. The cytologic smears contained diffuse small neoplastic cells larger than lymphocytes with dense, pyknotic nuclei and extremely scanty cytoplasm. Apparently viable large tumor cells have vesicular nuclei with granular, sometimes very coarse chromatin. The characteristic cytologic features of small cell carcinoma as compared to malignant lymphoma were as follows.: 1) small cells with dense pyknotic nuclei are evenly distributed in the background of apparently viable larger tumor cells, admixed with mature lymphocytes and phagocytic macrophages. 2) small loose aggregates of cells with nuclear melding are indicative of small cell carcinoma rather than non-Hodgkin's lymphoma. 3) the cytoplasmic and nuclear fragments of tumor necrosis are more dominant in the smears of small cell carcinoma. 4) nuclear membrane and nucleoli are generally indistinct in small cell carcinoma due to condensation of chromatin.

• International Journal of Oral Biology
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• v.40 no.1
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• pp.51-61
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• 2015

Shikonin, a major ingredient in the traditional Chinese herb Lithospermumerythrorhizon, exhibits multiple biological functions including antimicrobial, anti-inflammatory, and antitumor effects. It has recently been reported that shikonin displays antitumor properties in many cancers. This study was aimed to investigate whether shikonin could inhibit oral squamous carcinoma cell (OSCC) growth via mechanisms of apoptosis and cell cycle arrest. The effects of shikonin on the viability and growth of OSCC cell line, SCC25 cells were assessed by MTT assay and clonogenic assays, respectively. Hoechst staining and DNA electrophoresis indicated that the shikonin-treated SCC25 cells were undergoing apoptosis. Western blotting, immunocytochemistry, confocal microscopy, flow cytometry, MMP activity, and proteasome activity also supported the finding that shikonin induces apoptosis. Shikonin treatment of SCC25 cells resulted in a time- and dose-dependent decrease in cell viability, inhibition of cell growth, and increase in apoptotic cell death. The treated SCC25 cells showed several lines of apoptotic manifestation as follows: nuclear condensation; DNA fragmentation; reduced MMP and proteasome activity; decrease in DNA contents; release of cytochrome c into cytosol; translocation of AIF and DFF40 (CAD) onto the nuclei; a significant shift in Bax/Bcl-2 ratio; and activation of caspase-9, -7, -6, and -3, as well as PARP, lamin A/C, and DFF45 (ICAD). Shikonin treatment also resulted in down-regulation of the G1 cell cycle-related proteins and up-regulation of $p27^{KIP1}$. Taken together, our present findings demonstrate that shikonin strongly inhibits cell proliferation by modulating the expression of the G1 cell cycle-related proteins, and that it induces apoptosis via the proteasome, mitochondria, and caspase cascades in SCC25 cells.

• International Journal of Oral Biology
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• v.35 no.2
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• pp.51-60
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• 2010

Few studies have evaluated the apoptosis-inducing efficacy of NaF on cancer cells in vitro but there has been no previous investigation of the apoptotic effects of NaF on human oral squamous cell carcinoma cells. In this study, we have investigated the mechanisms underlying the apoptotic response to NaF treatment in the YD9 human squamous cell carcinoma cell line. The viability of YD9 cells and their growth inhibition were assessed by MTT and clonogenic assays, respectively. Hoechst staining, DNA electrophoresis and TUNEL staining were conducted to detect apoptosis. YD9 cells were treated with NaF, and western blotting, immunocytochemistry, confocal microscopy, FACScan flow cytometry, and MMP and proteasome activity assays were performed sequentially. The NaF treatment resulted in a time- and dose-dependent decrease in YD9 cell viability, a dose-dependent inhibition of cell growth, and the induction of apoptotic cell death. The apoptotic response of these cells was manifested by nuclear condensation, DNA fragmentation, the reduction of MMP and proteasome activity, a decreased DNA content, the release of cytochrome c into the cytosol, the translocation of AIF and DFF40 (CAD) into the nucleus, a significant shift of the Bax/Bcl-2 ratio, and the activation of caspase-9, caspase-3, PARP, Lamin A/C and DFF45 (ICAD). Furthermore, NaF treatment resulted in the downregulation of G1 cell cyclerelated proteins, and upregulation of p53 and the Cdk inhibitor $p27^{KIP1}$. Taken collectively, our present findings demonstrate that NaF strongly inhibits YD9 cell proliferation by modulating the expression of G1 cell cycle-related proteins and inducing apoptosis via mitochondrial and caspase pathways.

• YAKHAK HOEJI
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• v.41 no.1
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• pp.86-93
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• 1997

Even though both N-methyl-4-phenyl-1,2,5,6-tetrahydropyridine (MPTP) and 6-hydroxydopamine have been widely used to establish the experimental model for dopaminergic neuronal ce ll death. mechanisms underlying this phenomenon have not been firmly explored. To investigate how these dopaminergic neurotoxins induce neuronal cell death, murine dopaminergic neuronal cell line, MN9D cells were treated with various concentration of either 6-hydroxydopamine or active form of MPTP, N methyl-4-phenylpyridinium (MPP$^+$). Treatment of cells with 5-100 uM 6-hydroxydopamine resulted in apoptotic cell death whereas cell death induced by 5~50 uM MPP$^+$ was not demonstrated typical apoptotic characteristics such as cell shrinkage, apoptotic body and nuclear condensation. Cell death induced by 6-hydroxydopamine was partially blocked in the presence of antioxidants including soluble form of vitamin E or desferrioxamine suggesting that generation of oxidative stress may be associated with 6-hydroxydopamine-induced cell death in MN9D cells. In contrast, MPP$^+$-induced cell death was not blocked by treatment with any of antioxidants tested. As previously demonstrated that MPP$^+$ caused metabolic alterations such as glucose metabolism, removal of glucose from the medium partially inhibited MPP$^+$-induced cell death suggesting excessive cycles of glycolysis may be associated with MPP$^+$-induced cell death. Taken together, these studies demonstrate that two types of dopaminergic neurotoxins recruit distinct neuronal cell death pathways.

• Korean Journal of Biological Psychiatry
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• v.5 no.1
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• pp.66-70
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• 1998

Apoptosis is a form of cell death in which the cells shrink and exhibit nuclear chromatin condensation and DNA fragmentation, and yet maintain membrane integrity. Many lines of evidence have shown that brain neurons are vulnerable to degeneration by apoptosis. Also it has been suggested that apoptosis is one of the mechanism contributing neuronal loss in Alzheimer's disease(AD), since the conditions in the disease($A{\beta}$ peptide, oxidative stress, low energy metabolism) are the inducers that activate apoptosis. Indeed some neurons in vulnerable regions of the AD brain show DNA damage, chromatin condensation, and apoptic bodies. Consistently, mutations in AD causative genes(Amyloid precursor protein, Presenilin-1 and Presenilin- 2) increase $A{\beta}$ $peptide_{1-42}(A{\beta}_{1-42})$ and sensitize neuronal cell to apoposis. However, several lines of evidence have shown that the location of neuronal loss and $A{\beta}$ peptide deposition is not correlated in AD brain and transgenic mice brain over-expressing $A{\beta}_{1-42}$. Taken together, these data may indicated that $A{\beta}$ peptide(and other causative factors of AD) can interact with other cellular insults or risk factors to exacerbate pathological mechansim of AD through apoptosis. Thus, this review discusses possible role and mechanism of apoptosis in AD.

• Journal of Yeungnam Medical Science
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• v.12 no.2
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• pp.331-338
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• 1995

Laserthermia is a new method of local hyperthermia using fiber optic guided probe with computer controlled Nd-YAG laser system. We used a synthetic sapphire probe and allowed irradiation with contolled low power laser energy (less than 5W), in different thermal condition (temprature: 38.5~50 degrees C) for 10 minutes, in the normal brain tissue of 18 rabbits. In results, the histological changes of brain tissue was variable (myelin condensation, chromatin condensation, nuclear waving and palisading, RBC discoloration, cell necrosis) in microscopic findings after laser irradiation, but changing area was not occured proportionally in thermal condition level. Cell necrosis appears to over 44.5 degrees C and the distance was about 1.25 mm. This study, using computer controlled laserthermia system for interstitial local hyperthermia, may offer many advantages in the experimental treatment and clinical management of tumor. Minimizing normal tissue damage is now being developed.

• Korean Journal of Agricultural Science
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• v.49 no.2
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• pp.227-237
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• 2022

Amyloid beta (Aβ) is produced from an amyloid precursor protein by the activation of the amyloidogenic pathway, and it is widely known to cause Alzheimer's disease (AD). In this study, we investigated the neuroprotective effects of three flavonoids, quercitrin, isoquercitrin, and afzelin, from Acer okamotoanum against Aβ-induced neurotoxicity in SH-SY5Y neuronal cells. Aβ_25-35 treatments resulted in decreased cell viability and increased levels of nuclei condensation and fragmentation. However, an isoquercitrin treatment dose-dependently increased cell viability and decreased nuclei condensation and fragmentation levels. SH-SY5Y cells treated with Aβ_25-35 showed increased reactive oxygen species (ROS) production compared to that from cells not treated with Aβ_25-35. However, treatment with the three flavonoids significantly inhibited ROS production compared to an Aβ_25-35-treated control group, indicating that the three flavonoids blocked neuronal oxidative stress. For a closer examination of the neuroprotective mechanisms, we measured the expressions of the non-amyloidogenic pathway-related proteins of a disintegrin and metalloprotease 10 (ADAM10) and the tumor necrosis factor-α converting enzyme (TACE). An isoquercitrin treatment enhanced the expressions of ADAM10 compared to the control group. In addition, the three flavonoids activated the non-amyloidogenic pathway via the upregulation of TACE. In conclusion, we demonstrated neuroprotective effects of three flavonoids from A. okamotoanum, in particular isoquercitrin, on neurotoxicity by the regulation of the non-amyloidogenic pathway in Aβ_25-35-treated SH-SY5Y cells. Therefore, we suggest that flavonoids from A. okamotoanum may have some potential as therapeutics of AD.

• The Journal of Korean Obstetrics and Gynecology
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• v.20 no.2
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• pp.139-154
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• 2007

Purpose: This study examined the Cell differentiation and the anti-oxidative effect of Dioscoreae Rhizoma on HeLa cells. We are interested in whether Dioscoreae Rhizoma is capable of causing apoptosis processes on HeLa cell, and whether cotreatment of NCS with Dioscoreae Rhizoma reduces cell viability. Methods: We used aqueous extract to treat HeLa cell with different concentrations treated with a water or a MeOH extract of Dioscoreae Rhizoma (0, x10, x20, x40, x80). The MTT (3, (4, 5-dimethyl-thiazol) 2, 5-diphenyl-tetraxolium bromide) reduction assay was employed to quantify the differences in cell activity and viability. Cells were stained with DAPI and visualized by fluorescent Microscope. The caspase-3, Bcl-2, PARP, p53 expression level was monitored using western-blotting techniques. The patterns of the changes in expression were scanned and analyzed. Results: The survival rate of cells treated with Dioscoreae Rhizoma extracts increased by 20% at specific concentration. The other side Dioscoreae Rhizoma extracts induced apoptotic features including chromatin condensation and fragmentation. And Dioscoreae Rhizoma extracts increased the expression of caspase-3, p53 and the cleavage of PARP protein. However, co-treatment with Dioscoreae Rhizoma with NCS attenuated the activations of p53 and PARP protein that were mediated by NCS treatment alone. This is indicated Dioscoreae batatas extracts attenuated cytotoxicity induced by oxidative agents including NCS. Conclusion: Our results suggest Dioscoreae Rhizoma extracts induce cell differentiation or apoptosis connected with concentration. Further elucidation of concentration of Dioscoreae Rhizoma awaits many other biochemical investigative studies.

• Toxicological Research
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• v.21 no.4
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• pp.355-360
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• 2005

Epidemiological and laboratory studies provide insight into the anti-carcinogenic potential of garlic and its constituent compounds. Garlic is appealing as an anti-carcinogenic agent due to its ability to induce apoptosis in vitro. Diallyl disulfide (DADS) is one of the major components of garlic that used to determine inhibition of cell proliferation and induced apoptosis in human colon cell lines. In this study, human colorectal cancer cell lines (LOVO, HCT-116, SW-480) were exposed to DADS. The inhibitory effects of DADS dose level more than $50\;{\mu}M$ in the cell viability of all cell lines. Cell growth activity inhibits of human colon cancer cell lines. The inhibitory effects of DADS dose level more than $25\~50\;{\mu}M$ in the cell growth using MTT assay. We found that DADS may have the apoptosis action (chromatin condensation, DNA fragmentation) using DAPI staining and increased the expression of caspase-3 at the dose level more than $100\;{\mu}M$, decreased the expression level of $\beta-catenin$ at dose dependent in the western blotting. We suggest that DADS may have a potential candidate as cancer chemopreventive agents.