To analyze the effects of a new unknown peptide DEF on the growth of tumor cells, a fused polypeptide TAT-DV1-DEF was designed and synthesized. The lung adenocarcinoma cell line GLC-82 treated with TAT-DV1-DEF was analyzed with a cell counting kit 8, and the location of polypeptides in cells was observed under laser confocal microscopy. The efficiency of polypeptide transfection and changes in nuclear morphology were analyzed by flow cytometry and fluorescence microscopy, respectively. Finally, the mechanism of tumor cell growth inhibition was evaluated by Western blotting. We found that TAT-DV1-DEF could significantly inhibit the growth of the lung adenocarcinoma cell line GLC-82, but not the normal human embryonic kidney cell line HEK-293. Polypeptides were found to be mostly localized in the cytoplasm and some mitochondria. The efficiency of polypeptide transfection in the two cell types was approximately 99%. Apoptotic nuclei were observed under fluorescence microscopy upon treatment with polypeptides and DAPI staining. Western blot analyses indicated that the polypeptide inhibition of tumor cell growth was apoptosis dependent. In the present study, we demonstrated that fused polypeptides could induce apoptosis of the lung adenocarcinoma cell line GLC-82, indicating that the new unknown peptide DEF has antitumor effects.
Objectives : In this study, we investigate that Ulmi cortex extract contributes to growth inhibitory effect and anti-cancer activity on the HT-29 human colon cancer cells. Methods : Ulmi cortex was extracted from the leaves of the plant using water. The Ulmi cortex extract was treated to different concentrations for 24 hr. Growth inhibitory effect was analyzed by measuring FACS study and MTT assay. Cell cycle inhibition was confirmed by kinases assay. Cell apoptosis was confirmed by surveying caspases cascades activation using Western blot. Results : Exposure to Ulmi cortex extract (0.4mg/ml) results in an inhibitory effect on cell growth in HT-29 cells. Growth inhibition by Ulmi cortex extract in HT-29 cells was related with the inhibition of proliferation and induction of apoptosis. The Ulmi cortex extract induces G1-cell cycle arrest and DNA fragmentation in HT-29 cells. Furthermore, Ulmi cortex extract induces cell apoptosis through the activation of caspases-3 and PARP cleavage. Conclusion : Ulmi cortex extract induces apoptosis in human colon cancer cells, therefore, we suggest that Ulmi cortex extract can be used as a novel class of anti-cancer drugs.
Purpose: To evaluated the effect of the gambogic acid (GA), one of the effective components of Garcinia, in combination with a new multi-targeted oral medication, sunitinib (SU) on renal cancer cell proliferation in vitro and on tumor growth in vivo. Methods: After treatment with GA or SU, either alone or in combination, MTT and FACS analysis were used to examine cell viability and cycle distribution of the renal carcinoma cell lines 786-0 and Caki-1. Western blotting was employed to examine the expression of proteins related to the cell cycle and vascular formation. Furthermore, a xenograft model was applied to study the antitumor efficacy of SU or GA alone or in combination, with immunohistochemistry to detect expression of proteins related to xenograft growth and angiogenesis. Western blotting was used to examine NF-${\kappa}B$ signaling pathway elements in xenografts. Results: Treatment of 786-0 and Caki-1 cells with GA or SU resulted in decreased tumor cell proliferation, especially with joint use. Cells accumulated more strongly in the sub-G1 phase after joint treatment with GA and SU than treatment of GA and SU alone. Western blotting arrays showed 1 protein significantly upregulated, 2 proteins downregulated, and 2 proteins unchanged. Moreover, combined use of GA and SU inhibited the growth and angiogenesis of xenografts generated from Caki-1 significantly. Immunohistochemistry arrays showed downregulation of the expression of proteins promoting xenograft growth and angiogenesis, and Western blotting showed inhibition of the NF-${\kappa}B$ signaling pathway after treatment by GA alone and in combination with SU in xenografts. Conclusions: Our results show that the joint use of GA and SU can provide greater antitumor efficacy compared to either drug alone and thus may offer a new treatment strategy for renal cell carcinoma.
Journal of the Korean Society of Food Science and Nutrition
/
v.32
no.6
/
pp.931-936
/
2003
We investigated inhibitory effects of Hericium erinaceus on the growth of cancer cells and the expression of cell cycle regulators, cyclins. Anticancer effects of Hericium erinaceus extract and fractions against cancer cell lines including HepG2 and HT29 were investigated. The methanol extract, the hexane fraction, the chloroform fraction and the ethylacetate fraction of Hericiu erinacew inhibited growth of cancer cells but they had no effect on the cytotoxicity of normal human liver cells under the same conditions. As shown by western blot analysis, the expression of cyclin B1 known as cell cycle regulator was markedly decreased after treatment with Hericium erinaceus extract in HepG2 cells. These results suggest that antiproliferative effect of Hericaum erinaceus extract is associated with markedly decreased expression of cyclin B1.
Oncostatin M (OSM) is a multifunctional cellular regulator acting on a wide variety of cells, which has potential roles in the regulation of gene activation, cell survival, proliferation and differentiation. Previous studies have shown that OSM can induce morphological and/or functional differentiation and maturation of many tumor cells. However, the action of OSM on the induction of differentiation of human hepatocellular carcinoma (HCC) has not been reported. Here, we investigated the effects of different concentrations of OSM on human HCC cell line SMMC-7721 growth, proliferation, cell cycling, apoptosis and differentiation in vitro. Cell growth was determined via MTT assay, proliferation by cell cycle analysis, apoptosis by flow cytometry, morphology by transmission electronic microscopy, and cell function by detection of biochemical markers. Our results demonstrated that OSM strongly inhibited the growth of SMMC-7721 cells in a dose-dependent manner, associated with decreased clonogenicity. Cell cycle analysis revealed a decreased proportion of cells in S phase, with arrest at G0/G1. The apotosis rate was increased after OSM treatment compared to the control. These changes were associated with striking changes in cellular morphology, toward a more mature hepatic phenotype, accompanied by significant reduction of the expression of AFP and specific activity of ${\gamma}$-GT, with remarkable increase in secretion of albumin and ALP activity. Taken together, our findings indicate that OSM could induce the differentiation and reduce cell viability of SMMC-7721 cells, suggesting that differentiation therapy with OSM offers the opportunity for therapeutic intervention in HCC.
Journal of Physiology & Pathology in Korean Medicine
/
v.20
no.3
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pp.701-709
/
2006
The Rhus verniciflua Stokes (乾漆-RVS) has been used in traditional East Asia medicine for the therapy of gastritis, stomach cancer, although the mechanism for the biological activity is unclear. In the present study aims to investigate RVS extract contributes to growth inhibitory effect and it's the molecular mechanism on the human gastric cancer cells. AGS (gastric cancer cells) and RIEI (normal cells) were treated to different concentrations and periods of RVS extract $(10{\;}{\sim{{\;}100{\;}ug/mil)$. Growth inhibitory effect was analyzed by measuring FACS study and MTS assay. Cell cycle inhibition was confirmed by measuring CDK2 kinase activity by immunoprecipitation and kinase assay. And apoptosis was confirmed by surveying caspase cascades activation using a pan caspase inhibitor Exposure to RVS extract (50 ug/mll) resulted in a synergistic inhibitory effect on cell growth in AGS cells. Growth inhibition was related with the inhibition of proliferation and induction of apoptosis. The extract induces Gl -cell cycle arrest through the regulation of cyclins, the induction of p27kip1, and the decrease CDK2 kinase activity. And upregulated p27kip1 level is caused by protein stability increment by the reduction of S-phase kinase-associated protein 2 (Skp2), a key molecule related with p27kip1 ubiquitination and degradation, and do novo protein synthesis. Besides, 乾漆 extract induces apoptosis through the expression of Bax, poly(ADP-ribose) polymerase (PARP) and activation of caspase-3. RVS extract induces Gl -cell cycle arrest via accumulation of p27kip1 and apoptosis in human gastric cancer cells but not in normal cells, therefore we suggest that the extract can be used as a novel class of anti-cancer drugs.
Fumonisins are a family of mycotoxins produced by the fungus Fusarium moniliforme which is a common contaminant in corn. Fumonisins are potent inhibitors of sphingosine and sphinganine N-acyltransferase (ceramide synthase), key enzymes in sphingolipid metabolism. The purpose of this study was to provide the evidence that the elevated levels of free sphingoid bases (primarily sphinganine) and depletion of complex sphingolipids were closely related to the inhibition of cell growth in LLC-$PK_1$ cells exposed to fumonisin $B_1$$(\leq 35 {\mu}M)$. Concentrations of fumonisin $B_1$ between 10 and $35 {\mu}M$ were known to inhibit cell growth without cytotoxicity in $LLC-PK_1$ cells (Yoo et al. Toxicol. Appl. Pharmacol. 114, 9-15, 1992). Cells exposed to 35$\mu M$ fumonisin B$_1$ for 48 and 72 hr developed a fibroblast-like (elongated and spindle-shaped) appearance and were less confluent than normal cells. At between 24 and 48 hr after exposure to fumonisin $B_1$ cells were beginning to show the inhibition of cell growth and at 72 hr the number of viable cells in fumonisin-treated cultures was about 50% of concurrent control cultures. During the 24 hr lag period preceding inhibition of cell growth, the free sphinganine levels in cells exposed to $35 {\mu}M$ fumonisin $B_1$ were highly elevated (approximately 230 fold higher than normal cells). The elevated levels of free sphinganine were $435\pm14$$pmoles/{10^6}$ cells at 48 hr and approximately TEX>$333\pm11$$pmoles/{10^6}$ cells in cells exposed to $35{\mu}M$ fumonisin$B_1$ at 72 hr, while the levels of free sphinganine in normal cells were less than 2$pmoles/{10^6}$ cells. Under the same condition, depletion of intracellular complex sphingolipids as a consequence of fumonisin inhibition of de novo sphingolipid biosynthesis and turnover pathway was appeared. Content of free sphingold bases in dividing cells was more elevated than in confluent cells at 24-48 hr after cells were exposed to $20{\mu}M$ fumonisin $B_1$. The dividing cells were showing the inhibition of cell growth at 48-72 hr and $20{\mu}M$ fumonisin $B_1$. The results of this study support the hypothesis that the inhibition of cell growth is very well related to the disruption of sphingolipid metabolism in $LLC-PK_1$ cells.
Park, Gwang-Uk;Gang, Seok-Jin;Gwon, Ji-Hye;Kim, Jun-Beom;Yeo, Chan-Il;Lee, Yong-Tak
Proceedings of the Korean Vacuum Society Conference
/
2012.08a
/
pp.308-309
/
2012
One of the critical issues in the growth of multijunction solar cell is the formation of a highly doped Esaki interband tunnel diode which interconnects unit cells of different energy band gap. Small electrical and optical losses are the requirements of such tunnel diodes [1]. To satisfy these requirements, tens of nanometer thick gallium arsenide (GaAs) can be a proper candidate due to its high carrier concentration in low energy band gap. To obtain highly doped GaAs in molecular beam epitaxy, the temperatures of Si Knudsen cell (K-cell) for n-type GaAs and Be K-cell for p-type GaAs were controlled during GaAs epitaxial growth, and the growth rate is set to 1.75 A/s. As a result, the doping concentration of p-type and n-type GaAs increased up to $4.7{\times}10^{19}cm^{-3}$ and $6.2{\times}10^{18}cm^{-3}$, respectively. However, the obtained n-type doping concentration is not sufficient to form a properly operating tunnel diode which requires a doping concentration close to $1.0{\times}10^{19}cm^{-3}$ [2]. To enhance the n-type doping concentration, n-doped GaAs samples were grown with a lower growth rate ranging from 0.318 to 1.123 A/s at a Si K-cell temperature of $1,180^{\circ}C$. As shown in Fig. 1, the n-type doping concentration was increased to $7.7{\times}10^{18}cm^{-3}$ when the growth rate was decreased to 0.318 A/s. The p-type doping concentration also increased to $4.1{\times}10^{19}cm^{-3}$ with the decrease of growth rate to 0.318 A/s. Additionally, bulk resistance was also decreased in both the grown samples. However, a transmission line measurement performed on the n-type GaAs sample grown at the rate of 0.318 A/s showed an increased specific contact resistance of $6.62{\times}10^{-4}{\Omega}{\cdot}cm^{-2}$. This high value of contact resistance is not suitable for forming contacts and interfaces. The increased resistance is attributed to the excessively incorporated dopant during low growth rate. Further studies need to be carried out to evaluate the effect of excess dopants on the operation of tunnel diode.
In this work, effect of the culture medium composition on the fermentation process of Clostridium ljungdahlii, which is acetogenic bacteria to product ethanol from synthesis gas, was examined to improve the microbial growth and ethanol production. Components of the culture medium such as yeast extract, fructose, $NH_4Cl$, and $K_2HPO_4$ were selected as influence factors for the cell growth and ethanol production. As the concentration of yeast extract increased, both of the cell growth and ethanol production increased. And the ethanol productivity was the highest at an yeast extract of 0.05 g/L, which is lower than that of base medium. As the concentration of fructose increased, the cell growth increased, but the ethanol production decreased when the concentration of fructose was higher than that of base medium (5 g/L). In an experiment with the yeast extract of 5 g/L, produced ethanol concentration was the highest (0.297 g/L) when fructose concentration was 5 g/L, however, the specific ethanol productivity was higher (0.281 g/g DCW) when the fructose was not added due to very low cell mass. The cell growth and ethanol production were not significantly influenced by $NH_4Cl$ concentration, however the growth inhibition was observed at a 30 g/L of $NH_4Cl$. When the concentration of $K_2HPO_4$ increased, both of the cell growth and ethanol production increased. In experiments with $NH_4Cl$ and $K_2HPO_4$, specific ethanol productivities were higher when the low concentration of yeast extract was used.
This study was conducted to examine the effects of plug tray cell size and growth media on good seedling production of Gerbera hybrida Hort. Seedlings were grown for 60 days in 50, 72, 128, 162 cell trays contanning perlite, cocopeat and perlite+cocopeat(1:1, v/v). Perlite showed higher bulk density than cocopeat and perlite+cocopeat. Total porosity was greater in perlite, cocopeat and perlite+cocopeat in order. Cocopeat showed the highest water holding capacity. Number of leaves were greatest in 128 cell tray containing cocopeat. Leaf area was greatest in 50 cell tray containing cocopeat. Seedling growth was also better in plug tray of bigger cell size. Seedling growth of fresh and dry weight of shoot and root was much better in the growth media of perlite+cocopeat.
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