Background: DNA content analysis of human solid tumor is now widely performed by flow cytometric study. One of the most interesting and potentially important observation in this field is that proliferative activity(S-Phase fraction of cell cycle) may profoundly affect prognosis. Method: S-Phase fraction(SPF) have been measured by flow cytometric method using tumor cells isolated from paraffin embedded tissue. To evaluate the prognostic significance, SPF of squamous lung cancer cell was assessed in 21 patients who died without any specific treatment. Results: 1) Mean survival time of squamous lung cancer patients was 225(${\pm}162$) days. Survival time were shortened, when TNM stage and PS scale were advanced. 2) Mean value of SPF of squamous lung cancer patients was 23.4(${\pm}11.3$)%. SPF had nothing to do with advance of TNM stage and PS scale. 3) Mean survival time of high SPF group(more than 20% of cell proliferation cycle) and low SPF group were 153(${\pm}99$) days and 342(${\pm}180$) days(p<0.01). In each identical TNM stage and PS scale, there were also statistic significant differences in mean survival time between high and low SPF group. Conclusion: On multivariate analysis including TNM stage and performance status, SPF was the significant and independent prognostic factor in the primary squamous lung cancer patients group.
Esculetin, a coumarin compound, has been known to inhibit proliferation and induce apoptosis in several types of human cancer cells. However, the molecular mechanisms involved in esculetin-induced apoptosis are still uncharacterized in human leukemia cells. In this study, we have investigated whether esculetin exerts anti-proliferative and apoptotic effects on human leukemia U937 cells. It was found that esculetin could inhibit cell viability in a time-dependent manner, which was associated with the induction of apoptotic cell death such as increased populations of apoptotic- sub G1 phase. Apoptosis of U937 cells by esculetin was associated with an inhibition of Bcl-2/Bax binding activity, formation of tBid, down-regulation of X-linked inhibitor of apoptotic protein (XIAP) expression, and up-regulation of death receptor 4 (DR4) and FasL expression. Esculetin treatment also induced the degradation of ${\beta}$-catenin and DNA fragmentation factor 45/inhibitor of caspase-activated DNase (DFF45/ICAD). Furthermore, a caspase-3 specific inhibitor, z-DEVD-fmk, significantly inhibited sub-G1 phase DNA content, morphological changes and degradation of ${\beta}$-catenin and DEE45/ICAD. These results indicated that a key regulator in esculetin-induced apoptosis was caspase-3 in human leukemia U937 cells.
5-fluorouracil (5-FU), a pyrimidine analog, is a widely used anticancer drug, which works through irreversible inhibition of thymidylate synthase. In the present study, it was investigated the anti-proliferative effects and molecular mechanisms of 5-FU using Ewing's Sarcoma CHP-100 Cells. The present data indicated that treatment of 5-FU to CHP-100 cells induced a G1 phase arrest of the cell cycle in a time-dependent manner. 5-FU-induced G1 arrest was correlated with the accumulation of the hypophosphorylated form of the retinoblastoma protein (pRB) and association of pRB with the transcription factors E2F-1 and E2F-4. Although 5-FU treatment did affect the levels of cyclin-dependent kinases, the levels of cyclin A and B were markedly down-regulated as compared with the untreated control group. In addition, 5-FU-induced G1 arrest of CHP-100 cells was also associated with the induction of apoptosis, as determined by apoptotic cell morphologies, degradation of poly(ADP-ribose) polymerase and Annexin V staining. Furthermore, 5-FU induced the loss of mitochondrial membrane potential with up-regulated pro-apoptotic Bax expression, down-regulated anti-apoptotic Bcl-2 expression and cytochrome c release from mitochondria to cytosol. Collectively, the data suggest that 5-FU is effective in inducing cell growth reduction and apoptosis, in part, by reducing phosphorylation of pRB and activating mitochondrial dysfunction in CHP-100 cells.
Park, Dong-Wook;Yang, Hyun-Won;Kwon, Hyuek-Chan;Hwang, Kyung-Joo;Yoo, Jung-Hyun;Lee, Chi-Hyeong;Kim, Sei-Kwang;Cho, Dong-Jea;Oh, Kie-Suk
Clinical and Experimental Reproductive Medicine
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v.25
no.1
/
pp.25-33
/
1998
Cyclooxygenase (COX) is an enzyme involved in the conversion of arachidonic acid to prostaglandins (PGs), and exists in two forms, COX-1 and COX-2. COX has been reported to be involved in early implantation by secretion of PGs which causes permeability of vessels and reaction of decidual cells around the implantation site. Recently, in mice and sheep studies, COX-1 and COX-2 expression in the endometrium has been reported to be different according to implantation and stages of the estrous cycle, but expression of COX-1 and COX-2 in human endometrium during the menstrual cycle has not yet been established. The purpose of this study was to observe the variances of COX-1 and COX-2 expression by immunohistochemical staining in endometrial samples obtained from human hysterectomy specimens and biopsies of women of reproductive age according to different stages of the menstrual cycle. Also, we attempted to observe COX-1 and COX-2 expression in the epithelial and stromal cells of the endometrium obtained during the mid-secretory phase, which were cultured separately. COX-2 showed a cyclic pattern of expression according to the different stages of the menstrual cycle and was strongly expressed particularly at the mid-secretory phase which corresponds to the time of implantation. However, COX-1 tended to be increased in the early proliferative, and mid- and late secretory phases, but was also expressed in the whole menstrual cycle showing no particular pattern. In the separately cultured cells COX-1 was expressed in epithilial cells and COX-2 in the stromal cells. The above results suggest that since COX-2 is expressed at the same time as implantation and cultured cells display a specific secretory pattern, COX-2 has inductive endocrine enzyme properties and has an important effect on endometrial cells during implantation. Also, COX-2 expression in endometrial cells may be utilized as a useful marker of endometrial maturation.
Pituitary tumors are usually benign but can occasionally exhibit hormonal and proliferative behaviors. Dysregulation of the G1/S restriction point largely contributes to the over-proliferation of pituitary tumor cells. F-box protein S-phase kinase-interacting protein-2 (SKP2) reportedly targets and inhibits the expression of $p27^{Kip1}$, a well-known negative regulator of G1 cell cycle progression. In this study, SKP2 expression was found to be upregulated while $p27^{Kip1}$ expression was determined to be downregulated in rat and human pituitary tumor cells. Furthermore, SKP2 knockdown induced upregulation of $p27^{Kip1}$ and cell growth inhibition in rat and human pituitary tumor cells, while SKP2overexpression elicited opposite effects on $p27^{Kip1}$ expression and cell growth. The expression of microRNA-186 (miR-186) was reported to be reduced in pituitary tumors. Online tools predicted SKP2 to be a direct downstream target of miR-186, which was further confirmed by luciferase reporter gene assays. Moreover, miR-186 could modulate the cell proliferation and $p27^{Kip1}$-mediated cell cycle alternation of rat and human pituitary tumor cells through SKP2. As further confirmation of these findings, miR-186 and $p27^{Kip1}$ expression were downregulated, while SKP2 expression was upregulated in human pituitary tumor tissue samples; thus, SKP2 expression negatively correlated with miR-186 and $p27^{Kip1}$ expression. In contrast, miR-186 expression positively associated with $p27^{Kip1}$ expression. Taken together, we discovered a novel mechanism by which miR-186/SKP2 axis modulates pituitary tumor cell proliferation through $p27^{Kip1}$-mediated cell cycle alternation.
Hanna Lee;Ok-Yi Jeong;Hee Jin Park;Sung-Lim Lee;Eun-yeong Bok;Mingyo Kim;Young Sun Suh;Yun-Hong Cheon;Hyun-Ok Kim;Suhee Kim;Sung Hak Chun;Jung Min Park;Young Jin Lee;Sang-Il Lee
IMMUNE NETWORK
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v.23
no.6
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pp.45.1-45.22
/
2023
Interstitial lung disease (ILD) involves persistent inflammation and fibrosis, leading to respiratory failure and even death. Adult tissue-derived mesenchymal stem cells (MSCs) show potential in ILD therapeutics but obtaining an adequate quantity of cells for drug application is difficult. Daewoong Pharmaceutical's MSCs (DW-MSCs) derived from embryonic stem cells sustain a high proliferative capacity following long-term culture and expansion. The aim of this study was to investigate the therapeutic potential of DW-MSCs in experimental mouse models of ILD. DW-MSCs were expanded up to 12 passages for in vivo application in bleomycin-induced pulmonary fibrosis and collagen-induced connective tissue disease-ILD mouse models. We assessed lung inflammation and fibrosis, lung tissue immune cells, fibrosis-related gene/protein expression, apoptosis and mitochondrial function of alveolar epithelial cells, and mitochondrial transfer ability. Intravenous administration of DWMSCs consistently improved lung fibrosis and reduced inflammatory and fibrotic markers expression in both models across various disease stages. The therapeutic effect of DW-MSCs was comparable to that following daily oral administration of nintedanib or pirfenidone. Mechanistically, DW-MSCs exhibited immunomodulatory effects by reducing the number of B cells during the early phase and increasing the ratio of Tregs to Th17 cells during the late phase of bleomycin-induced pulmonary fibrosis. Furthermore, DW-MSCs exhibited anti-apoptotic effects, increased cell viability, and improved mitochondrial respiration in alveolar epithelial cells by transferring their mitochondria to alveolar epithelial cells. Our findings indicate the strong potential of DW-MSCs in the treatment of ILD owing to their high efficacy and immunomodulatory and anti-apoptotic effects.
Seo, Bo-Yong;Kim, Young-Min;Choi, Jae-Won;Yun, Mi-Jung;Jeon, Young-Chan;Jeong, Chang-Mo;Kim, Gyu-Cheon;Huh, Jung-Bo
The Journal of Korean Academy of Prosthodontics
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v.53
no.1
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pp.9-18
/
2015
Purpose: The purpose of this study is to examine characteristics of implant surface with RBM and anodizing treatments, and to evaluate the responses of osteoblast-like cell (MG-63 cell). Materials and methods: Grade IV titanium disks were fabricated (Diameter 10 mm, thickness 3 mm). Anodizing treatment (ASD) group, RBM and anodizing treatment (RBM/ASD) group, control (machined surface) group were divided. In this study, osteoblast-like cell was used for experiments. The experiments consist of surface characteristics evaluation by FE-SEM images, energy dispersive spectroscopy and stereo-SEM. In order to evaluate cell adhesion evaluation by crystal violet assay and observe cells form by confocal laser microscopy. To assess cell proliferation by XTT assay, cell differentiation by RT-PCR and mineralization by Alizarin red S stain assay. ELISA analyzer was used for Quantitative evaluation. Comparative analysis was run by one-way ANOVA (SPSS version 18.0). Differences were considered statistically significant at P<.05. Results: In ASD group and RBM/ASD group, the surface shape of the crater was observed and components of oxygen and phosphate ions in comparison with the control group were detected. The surface average roughness was obtained $0.08{\pm}0.04{\mu}m$ in the control group, $0.52{\pm}0.14{\mu}m$ in ASD group and $1.45{\pm}0.25{\mu}m$ in RBM/ASD group. In cell response experiments, ASD group and RBM/ASD group were significantly higher values than control group in cell adhesion and mineralization phase, control group was the highest values in the proliferative phase. In RT-PCR experiments, RBM/ASD group was showed higher ALP activity than other groups. RBM/ASD group in comparison with ASD group was significantly higher value for cell adhesion and proliferation phase. Conclusion: In the limitation of this study, we are concluded that the surface treatment with RBM/ASD seems more effective than ASD alone or machined surface on cellular response.
In this study, hatched male broiler chicks(Ross) were fed on a basal diet and LPS was administered via intraperitoneal injection three times every other day, on the 9th, 11th and 13th days of the experiment, and then PBMC and splenocytes were isolated on day 14. The degree of alama blue reduction was evaluated at 4, 24, 48, 96 and 120 h in the splenocytes, and at 4, 8, 12, 24 and 48 h for PBMC of incubation after the addition of alama blue solution to the media. The cell numbers used in this experiment were 103, 104 and 105 cells per well, and the con A levels were 0.0, 1.0, 5.0, and 10.0 ㎍ per ml of medium. 1. The degree of alama blue reduction was found to increase in a linear fashion with increasing incubation time and cell numbers, for both splenocytes and PBMC. 2. During acute phase response, the degree to which alama blue was reduced was significantly elevated (p<0.05) at an incubation time of 24 hr for the splenocytes, 4 hr for PBMC, and a cell number of 105 cells per well, respectively. 3. The raised reduction of alama blue to control was linear with Con A levels in medium, and higher reduction in Con A 10.0 ㎍ relative to 1.0 or 5.0 ㎍ in ml medium was shown 4. The medium with autologous serum evidenced a significantly (p<0.05) higher reduction of alama blue relative to FBS. 5. Splenocytes and PBMC from the LPS-injected birds evidenced significantly higher levels of alama blue reduction regardless of incubation time, number of cells, level of Con A added, or serum type, as compared with what was observed in normal birds. The results indicated that the assay conditions for proliferative activity using the alama blue method in birds in which the acute phase response had been activated via intraperitoneal LPS injection requires 4 hrs of incubation for PBMC, 24 hrs of incubation for splenocytes, and 10㎍ of Con A per ml of medium.
Objective: The purpose of this study was to evaluate the correlation between the expression pattern of vascular endothelial growth factor (VEGF) in endometrium and the pathogenesis of endometriosis by investigating VEGF expression patterns and their difference between eutopic endometrium of patients with endometriosis and that of normal controls without endometriosis. Methods: Endometrial sections were obtained from 64 hysterectomy specimens from women under age of 40, who had undergone hysterectomies and had histological evidence of endometriosis, with stage 3 and 4 according to the revised American Society for Reproductive Medicine classification. As for controls, 37 sections were gained from women diagnosed as having cervical intraepithelial neoplasia (CIN) of the uterine cervix and without evidence of pelvic endometriosis or adenomyosis during their operation. The VEGF content was evaluated immunohistochemically in the eutopic endometrium from 64 patients with endometriosis and 37 normal controls. Histological semiquantitative score (H-score) was calculated and compared between study group and control group throughout the menstrual cycle. Results: There was no significant difference in the H-score of VEGF in the eutopic endometrium between patients with endometriosis and controls without endometriosis when compared according to the same phase of the cycle, although the H-score of the study group was significantly higher in the secretory phase than the proliferative phase. Conclusion: The VEGF expression in the eutopic endometrium of women with endometriosis was not different from that of women without endometriosis. This study suggests VEGF expression in eutopic endometrium is unlikely associated with the pathogenesis of endometriosis.
Park Tae Yeol;Park Cheol;Yoon Hwa Jung;Choi Yung Hyun;Ko Woo Shin
Journal of Physiology & Pathology in Korean Medicine
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v.18
no.5
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pp.1449-1455
/
2004
Bufonis venenum (dried toad venom; Chinese name, Chan su) is a traditional Chinese medicine obtained from the skin venom gland of the toad. It has long been used in treating arrhythmia and other heart diseases in China and other Asian countries. Additionally, Bufonis venenum has been reported to selectively inhibit the growth of various lines of human cancer cells. In the present study, it was examined the effects of extract of Bufonis venenum (EBV) on the growth of human bladder carcinoma cell line T24 in order to investigate the anti-proliferative mechanism and induction of apoptosis by EBV. Treatment of T24 cells to EBV resulted in the growth inhibition, morphological change and induction of apoptotic cell death in a dose-dependent manner. Flow cytometric analysis revealed that EBV treatment caused G2/M phase arrest of the cell cycle and down-regulation of cyclin A, cyclin B1 and Cdc2, which was associated with a marked up-regulation of cyclin-dependent kinases (Cdks) inhibitor p21 (WAF1/CIP1) in a p53-independent manner. The Cdc25C expression was also significantly inhibited by EBV treatment, however Wee1 kinase expression was not affected. The induction of apoptotic cell death by EBV was connected with down-regulation of anti-apoptotic Bcl-XS/L expression without alteration pro-apoptotic Bax expression. Taken together, these findings suggest that EBV may be a potential chemotherapeutic agent for the control of human bladder carcinorma cells and further studies will be needed to identify the active compounds that confer the anti-cancer activity of EBV.
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