• Asian Pacific Journal of Cancer Prevention
• /
• v.13 no.11
• /
• pp.5339-5343
• /
• 2012

The induction of apoptosis in target cells is a key mechanism for most anti-tumor therapies. Bufalin is a cardiotonic steroid that has the potential to induce differentiation and apoptosis of tumor cells. Research on bufalin has so far mainly involved leukemia, prostate cancer, gastric cancer and liver cancer, and has been confined to in vitro studies. The bufadienolides bufalin and cinobufagin have been shown to induce apoptosis in a wide spectrum of cancer cell. The present article reviews the anticancer effects of bufalin. It induces apoptosis of lung cancer cells via the PI3K/Akt pathway and also suppressed the proliferation of human non-small cell lung cancer A549 cell line in a time and dose dependent manner. Bufalin, bufotalin and gamabufotalin, key bufadienolides, significantly sensitize human breast cancer cells with differing ER-alpha status to apoptosis induction by the TNF-related apoptosis-inducing ligand (TRAIL). In addition, bufadienolides induce prostate cancer cell apoptosis more significantly than that in breast epithelial cell lines. Similar effects have been observed with hepatocellular carcinoma (HCC) but the detailed molecular mechanisms of inducing apoptosis in this case are still unclear. Bufalin exerts profound effects on leukemia therapy in vitro. Results of multiple studies indicate that bufalin has marked anti-tumor activities through its ability to induce apoptosis. Large-scale randomized, double-blind, placebo or positive drug parallel controlled studies are now required to confirm the efficacy and apoptosis-inducing potential of bufalin in various cancers in the cliniucal setting.

• The Journal of Korean Obstetrics and Gynecology
• /
• v.31 no.4
• /
• pp.1-15
• /
• 2018

Objectives: Osteoporosis is considered a serious human disease. We developed an extract of mixed herbs containing root of Platycodon grandiflorum (ExMH-PGR), which is expected to be effective in preventing or treating osteoporosis. The aim of this study was to investigate the anti-osteoporotic effect of ExMH-PGR in osteoblastic MC3T3-E1 cells and osteoclastic RAW 264.7 cells. Methods: To examine the anti-osteoporotic effect of ExMH-PGR, osteoblast and osteoclast differentiation were induced and cultured with various concentrations of ExMH-PGR. Alkaline phosphatase (ALP) activity, collagen synthesis, osteocalcin production, and mineralization in MC3T3-E1 cells were analyzed. Tartrate-resistant acid phosphatase (TRAP) activity and the formation of actin ring in RAW 264.7 cells were analyzed. Results: ExMH-PGR at concentration up to $25{\mu}g/mL$ significantly increased ALP activity, collagen synthesis, osteocalcin production, and mineralization in MC3T3-E1 cells. ExMH-PGR at 50 to $200{\mu}g/mL$ significantly inhibited TRAP activity and the formation of actin ring in RAW 264.7 cells. Conclusions: These results demonstrate that ExMH-PGR stimulates osteoblastic activities and inhibits osteoclastic activities in in vitro systems, suggesting that ExMH-PGR might be considered as an anti-osteoporotic candidate for treatment of osteoporosis disease.

• Journal of Life Science
• /
• v.18 no.3
• /
• pp.336-343
• /
• 2008

Histone deacetylases (HDACs) inhibitors have emerged as the accessory therapeutic agents for various human cancers, since they can block the activity of specific HDACs, restore the expression of some tumor suppressor genes and induce cell differentiation, cell cycle arrest and apoptosis in vitro and in vivo. In the present study, we investigated that the effect of trichostatin A (TSA), an HDAC inhibitor, on the cell growth and apoptosis, and its effect on the nuclear factor-kappaB $(NF-{\kappa}B)$ activity in 267B1 human prostate epithelial cells. Exposure of 267B1 cells to TSA resulted in growth inhibition and apoptosis induction in and dose-dependent manners as measured by fluorescence microscopy, agarose gel electrophoresis and flow cytometry analysis. TSA treatment inhibited the levels of IAP family members such as c-IAP-1 and c-IAP-2 and induced the proteolytic activation of caspase-3, -8 and -9, which were associated with concomitant degradation of poly (ADP-ribose)-polymerase, ${\beta}-catenin$ and laminin B proteins. The increase in apoptosis by TSA was connected with the translocation of $NF-{\kappa}B$ from cytosol to nucleus, increase of the DNA binding as well as promoter activity of $NF-{\kappa}B$, and degradation of cytosolic inhibitor of KappaB $(I{\kappa}B)-{\alpha}$ protein. We therefore concluded that TSA demonstrated anti-proliferative and apoptosis-inducing effects on 267B1 cells in vitro, and that the activation of caspases and $NF-{\kappa}B$ may play important roles in its mechanism of action. Although further studies are needed, these findings provided important insights into the possible molecular mechanisms of the anti-cancer activity of TSA.

• Biomolecules & Therapeutics
• /
• v.20 no.1
• /
• pp.81-88
• /
• 2012

Histone deacetylases (HDACs) are enzymes involved in the remodelling of chromatin, and have a key role in the epigenetic regulation of gene expression. Histone deacetylase (HDAC) inhibitors are emerging as an exciting new class of potential anti-cancer agents. In recent years, a number of structurally diverse HDAC inhibitors have been identifi ed and these HDAC inhibitors induce growth arrest, differentiation and/or apoptosis of cancer cells in vitro and in vivo. However, the underlying molecular mechanisms remain unclear. This study aimed at investigating the anti-tumor activity of various HDAC inhibitors, IN-2001, using T47D human breast cancer cells. Moreover, the possible mechanism by which HDAC inhibitors exhibit anti-tumor activity was also explored. In estrogen receptor positive T47D cells, IN-2001, HDAC inhibitor showed anti-proliferative effects in dose-and time-dependent manner. In T47D human breast cancer cells showed anti-tumor activity of IN-2001 and the growth inhibitory effects of IN-2001 were related to the cell cycle arrest and induction of apoptosis. Flow cytometry studies revealed that IN-2001 showed accumulation of cells at $G_2$/M phase. At the same time, IN-2001 treatment time-dependently increased sub-$G_1$ population, representing apoptotic cells. IN-2001-mediated cell cycle arrest was associated with induction of cdk inhibitor expression. In T47D cells, IN-2001 as well as other HDAC inhibitors treatment significantly increased $p21^{WAF1}$ and $p27^{KIP1}$ expression. In addition, thymidylate synthase, an essential enzyme for DNA replication and repair, was down-regulated by IN-2001 and other HDAC inhibitors in the T47D human breast cancer cells. In summary, IN-2001 with a higher potency than other HDAC inhibitors induced growth inhibition, cell cycle arrest, and eventual apoptosis in human breast cancer possibly through modulation of cell cycle and apoptosis regulatory proteins, such as cdk inhibitors, cyclins, and thymidylate synthase.

• Journal of Embryo Transfer
• /
• v.31 no.3
• /
• pp.169-177
• /
• 2016

There is a growing interest in the application of primary hepatocytes for treatment of liver diseases in humans and for drug development. Several studies have focused on long-term survival and di-differentiation blocking of primary hepatocytes in an in vitro culture system. Therefore, the present study also aimed to optimize an in vitro culture system using primary rat hepatocytes. Primary rat hepatocytes from 6-week-old male Crl:CD rats were isolated using a modified two-step collagenase perfusion. Healthy $3.5{\times}10^6$ primary rat hepatocytes were seeded into a 2 dimensional (2D) culture in a 25T culture flask coated with collagen type I or into a 3D culture in a 125-ml spinner flask for 7 days. Production of plasma protein (ALB and TF), apoptosis (BAX and BCL2), and CYP (CYP3A1) related genes were compared between the 2D and 3D culture systems. The 3D culture system had an advantage over the 2D system because of the relatively high expression of ALB and low expression of BAX in the 3D system. However, the level of CYP3A1 did not improve in the 3D culture with and without the presence of a dexamethasone inducer. Therefore, 3D culture has an advantage for albumin production and primary rat hepatocyte survivability, but a low expression of CYP3A1 indicated that primary rat hepatocytes require a high-density culture for stress reduction by continuous flow.

• Journal of Veterinary Clinics
• /
• v.28 no.4
• /
• pp.399-402
• /
• 2011

In recent years, the mesenchymal stem cells (MSC) derived from various tissues have been widely tested for developing cell therapies, tissue repair and transplantation. Although there has been much interest in the immunomodulatory properties of MSC and their immunologic reactions following autologous, allogeneic and xenogenic transplantation of MSC in vivo, up to date, the expression of immunogenic markers, such as class I and II human leukocyte antigens (HLA), after differentiation of human umbilical cord blood (hUCB)-derived MSC has been poorly investigated and require extensive in vitro and in vivo testing. In this experiment, the expression of the HLA-ABC and HLA-DR on hUCB-derived MSC have been tested by immunocytochemical staining. The undifferentiated MSC were moderately stained for HLA-ABC but very weakly for HLA-DR. In order to investigate the inhibitory effect of allogeneic lymphocytes on proliferation of MSC, the MSC were cultured in the presence or absence of peripheral allogeneic lymphocytes stimulated with concanavalin A. The allogeneic lymphocytes did not significantly inhibit MSC proliferation. We conclude that hUCB-MSC expressed moderately class I HLA antigen while almost negatively class II HLA antigen. The MSC have an immunomodulatory effect which can suppress the allogeneic response of lymphocytes. These in vitro data suggest that allogeneic MSC derived from cord blood can be useful candidate for allogeneic cell therapy and transplantation without a major risk of rejection.

• Development and Reproduction
• /
• v.12 no.1
• /
• pp.67-76
• /
• 2008

Oxidative damage resulted from reactive oxygen species (ROS) is one of the main causes for the decrease of the viability during in vitro culture and cryopreservation process. This experiment was performed to determine the effects of antioxidants on the human hematopoietic stem cell (HSC) during cryopreservation procedure. HSCs cultured in vitro with or without antioxidants were frozen and then examined for stem cell potential after thawing. The cell viability of thawed HSC was increased in $\alpha$-tocopherol and ascorbic acid treatment group compared to control group ($62.7{\pm}8.0%$) and it was higher in 150 uM $\alpha$-tocopherol treatment group ($70.5{\pm}7.0%$). No significant difference was observed in the membrane integrity in all groups. In auto-differentiation rate, no significant difference was appeared in all groups, but was lower in 150 uM $\alpha$-tocopherol ($7.3{\pm}2.6%$) compared to control group ($10.1{\pm}1.6%$). These results demonstrate that treatment of antioxidants improves the efficiency of cryopreservation for HSC and $\alpha$-tocopherol may be considered effective antioxidant for the protective effect on HSC.

• Korean Journal of Plant Resources
• /
• v.12 no.2
• /
• pp.107-119
• /
• 1999

In this study, the induction regeneration of callus from immature embryo in trifoliata orange (Poncirus trifoliata RAFIN.) were accomplished. The embryogenic calli were induced from the immature embryo derived from seed when the calli were irradiated for 16hr at about 2,000 Lux in $\frac{1}{2}$ MS medium supplemented with 3% sucrose, and 44.4$\mu$M BA. Regeneration to whole plants was the most successful in MS medium containing 5.0$\mu$M BA. The yellowish callus was developed at 2 to 3 weeks of culture and the callus was changed from yellow to green at 5 to 6 weeks culture. In vitro regeneration was directly induced from embryogenic callus in MS medium containing 3% sucrose and 5.0$\mu$M BA. Multishoot was formed at 16 weeks culture. Moreover, when the root-formed plantlet was transplanted to soil, they grew to a whole plant. The compact cultured-cells were observed by light microscope after 4 weeks of cultivation and the embryogenic clumps were formed about the 5 weeks. At the same time, the neighboring cells were liquefied. In addition, differentiation of leaf and stem from the callus was observed after 12 weeks. The developed oil sacs and the profacicular cambium of the immature leaf were observed after 18 weeks. Therefore, we can see the considerable changes of cell arrangements according to the developmental stages of calli from trifoliata orange.

• Environmental Analysis Health and Toxicology
• /
• v.27
• /
• pp.10.1-10.7
• /
• 2012

Objectives: In recent years, a number of structurally diverse Histone deacetylase (HDAC) inhibitors have been identified and these HDAC inhibitors induce growth arrest, differentiation and/or apoptosis of cancer cells in vitro and in vivo. This study aimed at investigating the antitumor activity of newly synthesized HDAC inhibitor, 3-(4-dimethylamino phenyl)-N-hydroxy-2-propenamide (IN-2001) using human breast cancer cells. Methods: We have synthesized a new HDAC inhibitor, IN-2001, and cell proliferation inhibition assay with this chemical in estrogen receptor-positive human breast cancer MCF-7 cells. Cell cycle analysis on MCF-7 cells treated with IN-2001 was carried out by flow cytometry and gene expression was measured by RT-PCR. Results: In MCF-7 cells IN-2001 showed remarkable anti-proliferative effects in a dose- and time-dependent manner. In MCF-7 cells, IN-2001 showed a more potent growth inhibitory effect than that of suberoylanilide hydroxamic acid. These growth inhibitory effects were related to the cell cycle arrest and induction of apoptosis. IN-2001 showed accumulation of cells at $G_2$/M phase and of the sub-$G_1$ population in a time-dependent manner, representing apoptotic cells. IN-2001-mediated cell cycle arrest was associated with HDAC inhibitor-mediated induction of CDK inhibitor expression. In MCF-7 cells, IN-2001 significantly increased $p21^{WAF1}$ expression. Conclusions: In summary, cyclin-dependent kinase (CDK) induced growth inhibition, possibly through modulation of cell cycle and apoptosis regulatory proteins, such as CDK inhibitors, and cyclins. Taken together, these results provide an insight into the utility of HDAC inhibitors as a novel chemotherapeutic regime for hormone-sensitive and insensitive breast cancer.

• Korean Journal of Microbiology
• /
• v.47 no.4
• /
• pp.281-288
• /
• 2011

To screen downstream genes of Aspergillus nidulans MsnA showing amino acid sequence similarity to the zinc finger region of Msn2/4 stress response transcription factors in Saccharomyces cerevisiae, differentially expressed genes (DEG) in MsnA overexpressed or msnA null mutant strains compared to wild type have been isolated. The cognate gene IDs were identified by DNA sequencing of the selected DEGs. Among those, DEG6 was known as mstB encoding a putative monosaccharide transporter. Expression level of mstB mRNA was increased in MsnA overproducing strains and MsnA bound directly to the promoter region of mstB in vitro. MstB containing twelve transmembrane domains exhibited 80% of amino acid sequence identities to A. niger MstA a high-affinity monosaccharide transporter. A null mutant of mstB was phenotypically undistinguishable to wild type. On the other hand, forced overexpression of MstB caused the increased formation of sexual structure cleistothecia in 0.1% glucose condition where wild type showed almost no cleistothecia. This result implies that mstB is involved in transport of monosaccharide required for sexual differentiation.