• Asian Pacific Journal of Cancer Prevention
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• 제15권18호
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• pp.7919-7923
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• 2014

20(S)-Protopanaxadiol (PPD), a ginsenoside isolated from Pananx quinquefolium L., has been shown to inhibit growth and proliferation in several cancer cell lines. The aim of this study was to evaluate its anticancer activity in human breast cancer cells. MCF-7 cells were incubated with different concentrations of 20(S)-PPD and cytotoxicity was evaluated by MTT assay. Occurrence of apoptosis was detected by DAPI and Annexin V-FITC/PI double staining. Mitochondrial membrane potential was measured with Rhodamine 123. The Bcl-2 and Bax expression were determined by Western blot analysis. Caspase activity was measured by colorimetric assay. 20(S)-PPD dose-dependently inhibited cell proliferation in MCF-7 cells, with an $IC_{50}$ value of $33.3{\mu}M$ at 24h. MCF-7 cells treated with 20(S)-PPD presented typical apoptosis, as observed by morphological analysis in cell stained with DAPI. The percentages of annexin V-FITC positive cells were 8.92%, 17.8%, 24.5% and 30.5% in MCF-7 cells treated with 0, 15, 30 and $60{\mu}M$ of 20(S)-PPD, respectively. Moreover, 20(S)-PPD could induce mitochondrial membrane potential loss, up-regulate Bax expression and down-regulate Bcl-2 expression. These events paralleled activation of caspase-9, -3 and PARP cleavage. Apoptosis induced by 20(S)-PPD was blocked by z-VAD-fmk, a pan-caspase inhibitor, suggesting induction of caspase-mediated apoptotic cell death. In conclusion, the 20(S)-PPD investigated is able to inhibit cell proliferation and to induce cancer cell death by a caspase-mediated apoptosis pathway.

• Nutrition Research and Practice
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• 제8권4호
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• pp.377-385
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• 2014

BACKGROUND/OBJECTIVES: Abundant consumption of seaweeds in the diet is epidemiologically linked to the reduction in risk of developing cancer. In larger cases, however, identification of particular seaweeds that are accountable for these effects is still lacking, hindering the recognition of competent dietary-based chemo preventive approaches. The aim of this research was to establish the antiproliferative potency and angiosuppressive mode of action of Stoechospermum marginatum seaweed methanolic extract using various experimental models. MATERIALS/METHODS: Among the 15 seaweeds screened for antiproliferative activity against Ehrlich ascites tumor (EAT) cell line, Stoechospermum marginatum extract (SME) was found to be the most promising. Therefore, it was further investigated for its anti-proliferative activity in-vitro against choriocarcinoma (BeWo) and non-transformed Human embryonic kidney (HEK 293) cells, and for its anti-migratory/tube formation activity against HUVEC cells in-vitro. Subsequently, the angiosuppressive activity of S. marginatum was established by inhibition of angiogenesis in in-vivo (peritoneal angiogenesis and chorioallantoic membrane assay) and ex-vivo (rat cornea assay) models. RESULTS: Most brown seaweed extracts inhibited the proliferation of EAT cells, while green and red seaweed extracts were much less effective. According to the results, SME selectively inhibited proliferation of BeWo cells in-vitro in a dose-dependent manner, but had a lesser effect on HEK 293 cells. SME also suppressed the migration and tube formation of HUVEC cells in-vitro. In addition, SME was able to suppress VEGF-induced angiogenesis in the chorio allantoic membrane, rat cornea, and tumor induced angiogenesis in the peritoneum of EAT bearing mice. A decrease in the microvessel density count and CD31 antigen staining of treated mice peritoneum provided further evidence of its angiosuppressive activity. CONCLUSIONS: Altogether, the data underline that VEGF mediated angiogenesis is the target for the angiosuppressive action of SME and could potentially be useful in cancer prevention or treatment involving stimulated angiogenesis.

• Molecules and Cells
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• 제39권7호
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• pp.566-572
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• 2016

Lysosomes are cellular organelles containing diverse classes of catabolic enzymes that are implicated in diverse cellular processes including phagocytosis, autophagy, lipid transport, and aging. Lysosome-associated membrane proteins (LAMP-1 and LAMP-2) are major glycoproteins important for maintaining lysosomal integrity, pH, and catabolism. LAMP-1 and LAMP-2 are constitutively expressed in Salmonella-infected cells and are recruited to Salmonella-containing vacuoles (SCVs) as well as Salmonella- induced filaments (Sifs) that promote the survival and proliferation of the Salmonella. LAMP-3, also known as DC-LAMP/CD208, is a member of the LAMP family of proteins, but its role during Salmonella infection remains unclear. DNA microarray analysis identified LAMP-3 as one of the genes responding to LPS stimulation in THP-1 macrophage cells. Subsequent analyses reveal that LPS and Salmonella induced the expression of LAMP-3 at both the transcriptional and translational levels. Confocal Super resolution N-SIM imaging revealed that LAMP-3, like LAMP-2, shifts its localization from the cell surface to alongside Salmonella. Knockdown of LAMP-3 by specific siRNAs decreased the number of Salmonella recovered from the infected cells. Therefore, we conclude that LAMP-3 is induced by Salmonella infection and recruited to the Salmonella pathogen for intracellular proliferation.

• Molecules and Cells
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• 제39권7호
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• pp.536-542
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• 2016

Interleukin-17A is a member of the IL-17 family, and is known as CTLA8 in the mouse. It is produced by T lymphocytes and NK cells and has proinflammatory roles, inducing cytokine and chemokine production. However, its role in tumor biology remains controversial. We investigated the effects of locally produced IL-17A by transferring the gene encoding it into CT26 colon cancer cells, either in a secretory or a membrane-bound form. Expression of the membrane-bound form on CT26 cells dramatically enhanced their proliferation in vitro. The enhanced growth was shown to be due to an increased rate of cell cycle progression: after synchronizing cells by adding and withdrawing colcemid, the rate of cell cycle progression in the cells expressing the membrane-bound form of IL-17A was much faster than that of the control cells. Both secretory and membrane-bound IL-17A induced the expression of Sca-1 in the cancer cells. When tumor clones were grafted into syngeneic BALB/c mice, the tumor clones expressing the membrane-bound form IL-17A grew rapidly; those expressing the secretory form also grew faster than the wild type CT26 cells, but slower than the clones expressing the membrane-bound form. These results indicate that IL-17A promotes tumorigenicity by enhancing cell cycle progression. This finding should be considered in treating tumors and immune-related diseases.

• 한국산학기술학회논문지
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• 제17권3호
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• pp.127-135
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• 2016

Two-pore 도메인 포타슘 채널(two-pore domain $K^+$ channel, K2P channel)은 세포내 pH, 생리 활성 지질, 신경 전달 물질과 같은 생리학적 자극의 표적이며 안정막전압(resting membrane potential)을 설정하는 것으로 알려져 있다. 일부 유형의 K2P 채널들은 세포 사멸 및 종양 형성 등에서 중요한 역할을 한다. K2P 채널 중 TREK2 채널의 길항제는 보고되지 않았다. 본 연구의 목적은 TREK2 채널을 과발현시킨 HEK293 세포(HEKT2)에서 플라보노이드에 의해 TREK2 채널이 억제되는지 그리고 HEKT2 세포의 증식이 플라보노이드에 의해 영향을 받는지 알아보고자 하였다. 전기생리학적 전류는 단일 채널 patch clamp 방법을 사용하여 기록하였고 세포 증식은 XTT 에세이방법을 이용하여 측정하였다. HEKT2 세포에서 전기생리학적 TREK2 채널 활성도는 에피갈로카테킨-3-갈레이트(EGCG) 및 케르세틴과 같은 플라보노이드에 의해 각각 $91.5{\pm}13.1%$(n=5), $82.2{\pm}13.7%$(n=5)까지 억제되었다. 반면, EGCG 유사체인 에피카테킨(EC)는 TREK2 단일 채널 활성도에 현저한 억제 효과는 없었다. 또한 HEKT2 세포에서 세포 증식이 EGCG에 의해 $69.4{\pm}14.0%$(n=4)까지 감소되었음을 확인하였다. 결과로부터 EGCG와 케르세틴이 TREK2 채널 억제제임을 처음으로 확인하였고, EGCG만 HEKT2 세포의 증식을 감소시킨다는 결론을 얻었다. 본 연구의 결과는 EGCG 및 케르세틴이 TREK2 채널을 억제함으로써 막전압의 변화 유도와 세포 증식에 필요한 세포내 신호 변화의 시작을 트리거하는데 일차적으로 작동할 수 있음을 시사한다.

• Journal of Nutrition and Health
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• 제31권6호
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• pp.973-980
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• 1998

To investigate the effect of sea tangle on immune function in normal and diabetic states, 10-week old ICR mice were feed control(C) and sea tangle(5) diets containing 5%(w/w) cellulose and 13.6%(w/w) dry sea tangle for 4 weeks. After 4 weeks, three quarters of mice(CD and SD) were made diabetic by intramuscular injection of streptozotocin(150mg/kg bw). On the 4th day after diabetes was apparent by urinary glucose, one third of diabetic mire(CDG and SDG) were treated with glipizide(20mg/kg bw) and the other third(CDM and SDM) with metformin (500mg/kg bw) orally. Spleen weights of diabetic mice with no hypoglycemic drug treatment appeared to be higher in the sea tangle group(SD) than in control(CD), but were not different when drugs were administered. Data on splenocyte proliferation stimulated by lipopolysaccaride from Salmonella abortus equi(0.l$\mu\textrm{g}$/ml) showed that sea tangle increased mitogen response in normal mice(C group vs S group) and appeared to have the same effect in diabetic mice with or without drug treatment. Splenocyte proliferation induced by concanavalin A(0.1$\mu\textrm{g}$/ml) also showed similar results, although there were not statistically significant. Concentration of interleukin-2(IL-2) released from splenocytes of the S group seemed higher than from the C group, but the IL-2 concentrations were not different among six diabetic groups. Results of fatty acid compositions of splenocyte phospholipids showed that diabetes reduced arachidonic acid/linoleic acid ratios and that sea tangle intake and glipizide treatments increased contents of polyunsaturated fatty acids. It is concluded that dietary sea tangle has a positive effect on splenocyte proliferation under normal condition and could have the same effect under diabetic conditions. IL-2 appears to be one of factors mediating the effect but involvement of membrane fatty arid changes and other unknown factors needs lurker Investigation. (Korean J Nutrition 31(6) : 973-980, 1998)

• Molecules and Cells
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• 제38권6호
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• pp.518-527
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• 2015

Controlled release of medications remains the most convenient way to deliver drugs. In this study, we precipitated gold nanoparticles with quercetin. We loaded gold-quercetin into poly(DL-lactide-co-glycolide) nanoparticles (NQ) and tested the biological activity of NQ on HepG2 hepatocarcinoma cells to acquire the sustained release property. We determined by circular dichroism spectroscopy that NQ effectively caused conformational changes in DNA and modulated different proteins related to epigenetic modifications and c ell cycle control. The mitochondrial membrane potential (MMP), reactive oxygen species (ROS), cell cycle, apoptosis, DNA damage, and caspase 3 activity were analyzed by flow cytometry, and the expression profiles of different anti- and pro-apoptotic as well as epigenetic signals were studied by immunoblotting. A cytotoxicity assay indicated that NQ preferentially killed cancer cells, compared to normal cells. NQ interacted with HepG2 cell DNA and reduced histone deacetylases to control cell proliferation and arrest the cell cycle at the sub-G stage. Activities of cell cycle-related proteins, such as $p21^{WAF}$, cdk1, and pAkt, were modulated. NQ induced apoptosis in HepG2 cells by activating p53-ROS crosstalk and induces epigenetic modifications leading to inhibited proliferation and cell cycle arrest.

• 동의생리병리학회지
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• 제18권4호
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• pp.1147-1152
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• 2004

To investigate the anti-cancer effects of aqueous extract of Cheonkumwikyung-tang (CKWKT) on the growth of human lung carcinoma cell line A549, we performed various biochemical experiments such as the effects of CKWKT on the cell proliferation and viability, the morphological changes, the effects on expression of apoptosis and cell growth-regulatory gene products. Results obtained are as follow; CKWKT treatment declined the cell viability and proliferation of A549 cells in a concentration-dependent manner. The anti-proliferative effect by CKWKT treatment in A549 cells was associated with morphological changes such as membrane shrinking and cell rounding up. CKWKT treatment induced apoptotic cell death of A549 cells in a concentration-dependent manner, which was associated with inhibition and/or degradation of apoptotic target proteins such poly(ADP-ribose) polymerase, β-catenin and phospholipase C-γ1. Western blot analysis revealed that the levels cyclin-dependent kinase inhibitor p21 expression were induced by CKWKT treatment in A549 cells. Taken together, these findings suggest that CKWKT-induced inhibition of human lung cancer cell proliferation is associated with the induction of apoptotic cell death via regulation of several major growth regulatory gene products and CKWKT may have therapeutic potential in human lung cancer.

• 동의생리병리학회지
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• 제18권4호
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• pp.1102-1106
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• 2004

To investigate the anti-proliferative effects of an aqueous extract of Cordyceps militaris (AECM) on the growth of human lung carcinoma cell line A549, we performed various biochemical experiments such as the effects of AECM on the cell proliferation and viability, the morphological changes, the effects on expression of apoptosis and cell growth-regulatory gene products. Results obtained are as follow; AECM treatment declined the cell viability and proliferation of A549 cells in a concentration-dependent manner. The anti-proliferative effect by AECM treatment in A549 cells was associated with morphological changes such as membrane shrinking and cell rounding up. Taken together, these findings suggest that AECM-induced inhibition of human lung cancer cell proliferation is associated with the induction of apoptotic cell death via regulation of several major growth regulatory gene products, and C. militaris may have therapeutic potential in human lung cancer.

• Journal of Nutrition and Health
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• 제31권6호
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• pp.1076-1081
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• 1998

The purpose of this research was to isolate an active component on immunoytes from 70% MeOH extract of Codonopsis lanceolatae Radix(CLE). CLE was fractionated successively with hexane, methylene chloride, n-butyl alcohol and water, and then the water fraction was separated with molecularporous membrane tubing(m.w. 3,500). Each fraction(50mg/kg) was administered p.o. once a day for 7 days in BALB/c mice respectively. None of these fractions affected the apoptosis and mitochondrial transmembrane potential in thymocyte. Hexane and methylene chloride fractions decreased CD4$^{[-10]}$ CD8$^{+}$ single-positive cells, and the water fraction enhanced CD4$^{+}$ CD8$^{[-10]}$ single-positive cells in thymocyte. The proliferation of thymocytes was decreased by the fraction hexane, but was enhanced by the water fraction. Hexane, methylene chloride and butyl alcohol fractions suppressed the production of nitric oxide, which was not affected by the water fraction. Hexane and butyl alcohol fraction suppressed the phagocytic activity, but water fraction enhanced it. The components(m.w. 3,500 above) separated from the water fraction enhanced the proliferation of thymocyte, the population of CD4$^{+}$ CDB$^{[-10]}$ single-positive cells, and phagocytic activity in macrophage. These results suggest that the stimulative components of proliferation, TH population and phagorytc activity is in the water fraction, and the molecular weight is 3,500 above. (Korean J Nutrition 31(6) : 1076~1081, 1998) 1998)