• BMB Reports
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• v.49 no.1
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• pp.63-68
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• 2016

The serine/threonine kinase mTOR is essential for the phosphoinositide 3-kinases (PI3K) signaling pathway, and regulates the development and function of immune cells. Aberrant activation of mTOR signaling pathway is associated with many cancers including leukemia. Here, we report the contributions of mTOR signaling to growth of human leukemic cell lines and mouse T-cell acute leukemia (T-ALL) cells. Torin, an ATP-competitive mTOR inhibitor, was found to have both cytotoxic and cytostatic effects on U-937, THP-1, and RPMI-8226 cells, but not on Jurkat or K-562 cells. All cells were relatively resistant to rapamycin even with suppressed activity of mTOR complex 1. Growth of T-ALL cells induced by Notch1 was profoundly affected by torin partially due to increased expression of Bcl2l11 and Bbc3. Of note, activation of Akt or knockdown of FoxO1 mitigated the effect of mTOR inhibition on T-ALL cells. Our data provide insight on the effect of mTOR inhibitors on the survival and proliferation of leukemic cells, thus further improving our understanding on cell-context-dependent impacts of mTOR signaling. [BMB Reports 2016; 49(1): 63-68]

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.12
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• pp.1686-1692
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• 2009

Garlic oil (GAR, Allium sativum L.) has been studied as a feed additive to improve animal production performance and decrease methane emission in ruminants. The present study was designed to determine the possible effect of GAR on fatty acid composition and accumulation in animal fat tissue using a cell model. 3T3-L1 preadipocytes at $2{\times}10^{4}\;mL^{-1}$ were seeded to 24-well plates and allowed to proliferate to reach confluence. The cells were then treated with media containing 0, 2.5, 5, 10, 20 and 40 $\mu{g}$ $mL^{-1}$ of GAR during the differentiation period for 8 days. Media containing dexamethasone, methyl-isobutylxanthine and insulin was applied during the first 2 days of the early differentiation period. On day 8 sub-sets of the wells were stained with oil red-O and the remaining cells were harvested for determination of glycerol-3-phosphate dehydrogenase [EC 1.1.1.8] (GPDH) activity (n = 6) and cellular fatty acid concentration (n = 6). It was found that supplementation of GAR increased (p<0.05) the ratio of monounsaturated fatty acids/saturated fatty acids in the adipocytes and showed inhibitory effect (p<0.05) on the post-confluent proliferation. With relative low dosage, GAR (5-20 $\mu{g}$ $mL^{-1}$) increased (p<0.05) the GPDH activity without affecting the cellular fatty acid concentration, while a high dosage (40 $\mu{g}$ $mL^{-1}$) inhibited (p<0.05) fatty acid accumulation and decreased GPDH activity. Supplementation of GAR had an effect on cell post-confluent proliferation, differentiation and fatty acid accumulation. However, the effect may be diverse and depends on the dose applied.

• The Journal of Korean Medicine
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• v.19 no.1
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• pp.327-338
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• 1998

To investigate effect of water extract of PaIMuITang(PMT) on human cancer cell-lines and immunocytes, this research estimated proliferation of A431 cell line, KHOS-NP cell line, mouse thymocytes and mouse splenocytes, Nitric Oxide(NO) from macrophage, apoptosis and subpopulation of the mouse thymocytes. The results were obtained as follows; 1. PMT inhibited the. proliferation of A431 cell line, but it is not significant. 2. PMT inhibited the proliferation of KHOS-NP cell line, but it is not significant. 3. PMT stimulated the proliferation of mouse thymocytes, being compared Con A non-treated group. 4. PMT stimulated the proliferation of mouse splenocytes, being compared LPS treated group. 5. PMT l00g/mQ inhibited the production of NO from macrophages in vitro, being compared NPS IFN treated group. 6. PMT inhibited the production of NO from macrophages in vivo, being compared LPS|IFN treated group. 7. PMT accelerated the induction of apoptosis of the mouse thymocytes. 8. In subpopulation PMT decreased $T_H$ of the mouse thymocytes, but increased T /dT s of the mouse thymocytes.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.3
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• pp.419-427
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• 2016

There is a high association of heat shock on the alteration of energy and lipid metabolism. The alterations associated with thermal stress are composed of gene expression changes and adaptation through biochemical responses. Previous study showed that Angelica gigas Nakai (AGN) root extract promoted adipogenic differentiation in murine 3T3-L1 preadipocytes under the normal temperature condition. However, its effect in heat shocked 3T3-L1 cells has not been established. In this study, we investigated the effect of AGN root hot water extract in the adipogenic differentiation of murine 3T3-L1 preadipocytes following heat shock and its possible mechanism of action. Thermal stress procedure was executed within the same stage of preadipocyte confluence (G0) through incubation at $42^{\circ}C$ for one hour and then allowed to recover at normal incubation temperature of $37^{\circ}C$ for another hour before AGN treatment for both cell viability assay and Oil Red O. Cell viability assay showed that AGN was able to dose dependently (0 to $400{\mu}g/mL$) increase cell proliferation under normal incubation temperature and also was able to prevent cytotoxicity due to heat shock accompanied by cell proliferation. Confluent preadipocytes were subjected into heat shock procedure, recovery and then AGN treatment prior to stimulation with the differentiation solution. Heat shocked preadipocytes exhibited reduced differentiation as supported by decreased amount of lipid accumulation in Oil Red O staining and triglyceride measurement. However, those heat shocked preadipocytes that then were given AGN extract showed a dose dependent increase in lipid accumulation as shown by both evaluation procedures. In line with these results, real-time polymerase chain reaction (RT-PCR) and Western blot analysis showed that AGN increased adipogenic differentiation by upregulating heat shock protection related genes and proteins together with the adipogenic markers. These findings imply the potential of AGN in heat shock amelioration among 3T3-L1 preadipocytes through heat shock factor and proteins augmentation and enhanced adipogenic marker expression.

• Macromolecular Research
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• v.11 no.1
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• pp.42-46
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• 2003

Cell attachment and proliferation on the polymer films of triblock copolymer(ester-ether)s comprising po1y (L-1actide) (PLLA) and poly (oxyethylene-co-oxypropylene)(PN) were investigated using 3T3 fibroblasts. It was found that on the tissue culture polystyrene(TCPS) and the PLLA control film the cells could spread well while on the copolymer films the cells showed a rounded morphology without spreading and proliferated weakly. Especially, little cells proliferated on the films of copolymer having a LN composition of 20 wt%. While the water absorption of the copolymer films increased with increasing PN content, the contact angle against water of copolymer films immersed in aqueous medium was almost identical, being slightly lower than that of the PLLA film. These properties were compatible with the results of cell attachment. The in vitro hydrolysis of the films of triblock and multiblock type copolymers was faster with increasing PN content. The increased hydrolyzability, the flexibility and the decreased cell attachment suggested that these copolymers may have high potential as biodegradable materials for medical use.

• The Korea Journal of Herbology
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• v.29 no.4
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• pp.77-82
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• 2014

Objectives : The effects of ethanol extract of Plantago asiatica L. were investgated on adipocyte differentiation, lipopogenesis, lipolysis and apoptosis using differnentiated 3T3-L1 adipocytes. Methods : Plantago asiatica L. was extracted with ethanol (CCE). We carried on MTT assay for cell proliferation, Oil Red O staining for determination of cell differentiation and intracelluar adipogenesis. TUNEL staining assay for cell apoptosis, and Western blot analysis for measurement of pAMPK and pACC, $C/EBP{\alpha}$, $PPAR{\gamma}$ protein expressions were performed. Results : The addition of CCE up to 0.2 mg/ml into cell culture media showed no cytotoxicity. Treatment of 0.2 mg/ml CCE significantly inhibited differentiation in 3T3-L1 preadipocytes. Lipid accumulation of the CCE treated cells was decreased compared with that of control. Induction of cell apoptosis was increased in CCE treated cells compared with that of control. AMPK and ACC levels of the cells with 0.2 mg/ml CCE were led to phosphorylation and also expressions of $C/EBP{\alpha}$ and $PPAR{\gamma}$, as adipogenic transcription factors, were suppressed compared with those of control. Conclusions : Taken together, these results provide evidence that CCE has a regulatory role in lipid metabolism that is related to differentiation into adipocytes, adipogenesis and apoptosis.

• Preventive Nutrition and Food Science
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• v.12 no.1
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• pp.1-6
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• 2007

Colostrum basic proteins (CBP) (MW 1$\sim$30 kDa) were isolated from bovine colostrum using a series of ultrafiltration processes and their effects on osteoblast cell proliferation and bone metabolism were investigated in cell line and animal models. Treatments with CBP (1, 10, 100 $\mu$g/mL) dose-dependently increased cell proliferation of osteoblastic MC3T3 cells. Alkaline phosphatase activity, a marker of osteoblastic phenotype, in the cells was also increased after treatments with CBP in a dose-dependent manner. Significant increases in bone density were observed in femur of ovariectomized rats which were fed a diet with 1% and 10% CBP, compared to rats fed a normal diet. These results suggest that CBP may increase bone mass and density and be useful for the prevention of bone-related diseases.

• The Journal of Internal Korean Medicine
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• v.26 no.1
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• pp.93-106
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• 2005

Objectives : The aim of this study is to characterize the effect of Chungganhaeju-tang on $TGF-{\beta}l$-induced hepatic fibrosis. Materials and Methods : mRNA and protein expression levels of $TGF-{\beta}l$ in Chungganhaeju-tang treated HepG2 cells were compared to untreated cells using quantitative RT-PCR and ELISA assay, respectively. mRNA expression levels of the $TGF-{\beta}l$ signaling pathway genes $(T{\beta}R-I,\;T{\beta}R-II,\;Smad2,\;Smad3,\;Smad4,\;and\;PAI-1)$ and fibrosis-associated genes (CTGF, fibronectin, and collagen type $l{\alpha}$) were evaluated by quantitative RT-PCR. The effect of Chungganhaeju-tang on cell proliferation of T3891 human fibroblast was evaluated using $[^3H]Thymidine$ Incorporation Assay. Results : Inhibition of $TGF-{\beta}l$ mRNA expression and protein production was observed with treatment of Chungganhaeju-tang and seen to be dose and time dependent. Whereas $TGF-{\beta}l$-mediated induction of PAI-1 was suppressed with treatment of Chungganhaeju-tang, expression of the $TGF-{\beta}l$ signaling pathway genes such as $T{\beta}R-I$, $T{\beta}R-II$, Smad2, Smad3, and Smad4 was not affected. With treatment of Chungganhaeju-tang, inhibition of $TGF-{\beta}l$-induced cell proliferation of T3891 human fibroblast was observed, as well as abrogation of $TGF-{\beta}l$-mediated transcriptional up-regulation of CTGF, fibronectin, and collagen type $I{\alpha}$. Conclusion : This study strongly suggests that the liver cirrhosis-suppressive activity of Chungganhaeju-tang may be derived at least in part from its inhibitory effect on $TGF-{\beta}l$ functions, such as blockade of $TGF-{\beta}l$ stimulation of fibroblast cell proliferation and fibrosis-related gene expression as well as expression of $TGF-{\beta}l$ itself.

• Journal of Microbiology and Biotechnology
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• v.18 no.12
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• pp.1895-1902
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• 2008

Adipose tissue is an important endocrine organ involved in the control of whole body energy homeostasis and insulin sensitivity. Considering the increased incidence of obesity and obesity-related disorders, including diabetes, it is important to understand thoroughly the process of adipocyte differentiation and its control. Therefore, we performed a differential proteome mapping strategy using two-dimensional gel electrophoresis combined with peptide mass fingerprinting to identify intracellular proteins that are differentially expressed during adipose conversion of 3T3-L1 pre-adipocytes in response to an adipogenic cocktail. In the current study, we identified 46 differentially expressed proteins, 6 of which have not been addressed previously in 3T3-L1 cell differentiation. Notably, we found that phosphoribosyl pyrophosphate synthetase (PRPS), a regulator of cell proliferation, was preferentially expressed in pre-adipocytes than in fully differentiated adipocytes. In conclusion, our results provide valuable information for further understanding of the adipogenic process.

• Journal of Animal Science and Technology
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• v.51 no.5
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• pp.413-420
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• 2009

The research has been aimed to investigate the effect of different sera including fetal bovine serum (FBS), male bovine serum (MS), female bovine serum (FS), and castrated-male bovine serum (C-MS) on cell proliferation, follicular maturation and ovulation in vitro. Established cell lines and primary cells were cultured in the culture media supplemented with different sera and cells proliferation was observed by cell counting and MTT assay. The results indicated that cell proliferation was significantly different for different serum source. Proliferation of bovine and human myogenic satellite cells was highest in MS. In contrast, proliferation of breast cancer cells and immune cells were the highest in FS and FBS, respectively. There was no difference in the rate of follicular growth, whereas the rate of ovulation was higher in FBS and C-MS. Finally, the wound healing effect and cell proliferation of 3T3-L1 cells showed that wound healing was fastest in FS and cell proliferation was higher in MS. These results suggest the importance of an optimal serum selection in the experiments involving cell culture system, and gender-specific Hanwoo sera may be used as a substitute to FBS.