• Asian Pacific Journal of Cancer Prevention
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• 제15권13호
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• pp.5311-5316
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• 2014

Nuclear factor erythroid 2-related factor 2 (Nrf2) has been recognized as a transcription factor that controls mechanisms of cellular defense response by regulation of three classes of genes, including endogenous antioxidants, phase II detoxifying enzymes and transporters. Previous studies have revealed roles of Nrf2 in resistance to chemotherapeutic agents and high level expression of Nrf2 has been found in many types of cancer. At physiological concentrations, luteolin as a flavonoid compound can inhibit Nrf2 and sensitize cancer cells to chemotherapeutic agents. We reported luteolin loaded in phytosomes as an advanced nanoparticle carrier sensitized MDA-MB 231 cells to doxorubicin. In this study, we prepared nano phytosomes of luteolin to enhance the bioavailability of luteolin and improve passive targeting in breast cancer cells. Our results showed that cotreatment of cells with nano particles containing luteolin and doxorubicin resulted in the highest percentage cell death in MDA-MB 231cells (p<0.05). Furthermore, luteolin-loaded nanoparticles reduced Nrf2 gene expression at the mRNA level in cells to a greater extent than luteolin alone (p<0.05). Similarly, expression of downstream genes for Nrf2 including Ho1 and MDR1 were reduced significantly (p<0.05). Inhibition of Nrf-2 expression caused a marked increase in cancer cell death (p<0.05). Taken together, these results suggest that phytosome technology can improve the efficacy of chemotherapy by overcoming resistance and enhancing permeability of cancer cells to chemical agents and may thus be considered as a potential delivery system to improve therapeutic protocols for cancer patients.

• Asian Pacific Journal of Cancer Prevention
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• 제16권8호
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• pp.3267-3272
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• 2015

Purpose: To retrospective assess the potential predictors for relapse and create an effective clinical mode for surveillance after orchidectomy in clinical stage I non-seminomatous germ cell testicular tumors (CSI-NSGCTs). Materials and Methods: We analyzed data for CSI-NSGCTs patients with non-lymphatic vascular invasion, %ECa < 50% (percentage of embryonal carcinoma < 50%), and negative or declining tumor markers to their half-life following orchidectomy (defined as low-risk patients); these patients were recruited from four Chinese centers between January 1999 and October 2013. Patients were divided into active surveillance group and retroperitoneal lymph node dissection (RPLND) group according to different therapeutic methods after radical orchidectomy was performed. The disease-free survival rates (DFSR) and overall survival rates (OSR) of the two groups were compared by Kaplan-Meier analysis. Results: A total of 121 patients with CSI-NSGCT were collected from four centers, and 81 low-risk patients, including 54 with active surveillance and 27 with RPLND, were enrolled at last. The median follow-up duration was 66.2 (range 6-164) months in the RPLND group and 65.9 (range 8-179) months in the surveillance group. OSR was 100% in active surveillance and RPLND groups, and DFSR was 89.8% and 87.0%, respectively. No significant difference was observed between these two groups ($X_2=0.108$, P=0.743). No significant difference was observed between the patients with a low percentage of embryonal carcinoma (<50%) and those without embryonal carcinoma (87.0% and 91.9%, $X_2=0.154$, P=0.645). No treatment-related complications were observed in the active surveillance group whereas minor and major complications were observed in 13.0% and 26.1% of the RPLND group, respectively. Conclusions: Active surveillance resulted in similar DFSR and OSR compared with RPLND in our trial. Patients with low-risk CSI-NSGCTs could benefit from risk-adapted surveillance after these patients were subjected to radical orchidectomy.

• Asian Pacific Journal of Cancer Prevention
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• 제13권5호
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• pp.1809-1817
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• 2012

Background: Gastric cancer is frequently lethal despite aggressive multimodal therapies, and new treatment approaches are therefore needed. Retinoids are potential candidate drugs: they prevent cell differentiation, proliferation and malignant transformation in gastric cancer cell lines. They interact with nuclear retinoid receptors (the retinoic acid receptors [RARs] and retinoid X receptors [RXRs]), which function as transcription factors, each with three subclasses, ${\alpha}$, ${\beta}$ and ${\gamma}$. At present, little is known about retinoid expression and influence on prognosis in gastric cancers. Patients and Methods: We retrospectively analyzed the expression of the subtypes RARa, $RAR{\beta}$, $RAR{\gamma}$, RXRa, $RXR{\beta}$, $RXR{\gamma}$ by immunohistochemistry in 147 gastric cancers and 51 normal gastric epithelium tissues for whom clinical follow-up data were available and correlated the results with clinical characteristics. In addition, we quantified the expression of retinoid receptor mRNA using real-time PCR (RT-PCR) in another 6 gastric adenocarcinoma and 3 normal gastric tissues. From 2008 to 2010, 80 patients with gastric cancers were enrolled onto therapy with all-trans-retinoic acid (ATRA). Results: RARa, $RAR{\beta}$, $RAR{\gamma}$ and $RXR{\gamma}$ positively correlated with each other (p < 0.001) and demonstrated significantly lower levels in the carcinoma tissue sections (p < 0.01), with lower $RAR{\beta}$, $RAR{\gamma}$ and RXRa expression significantly related to advanced stages (p < =0.01). Tumors with poor histopathologic grade had lower levels of RARa and $RAR{\beta}$ in different histological types of gastric carcinoma (p < 0.01). Patients whose tumors exhibited low levels of RARa expression had significantly lower overall survival compared with patients who had higher expression levels of this receptor (p < 0.001, HR=0.42, 95.0% CI 0.24-0.73), and patients undergoing ATRA treatment had significantly longer median survival times (p = 0.007, HR=0.41, 95.0% CI 0.21-0.80). Conclusions: Retinoic acid receptors are frequently expressed in epithelial gastric cancer with a decreased tendency of expression and RARa may be an indicator of a positive prognosis. This study provides a molecular basis for the therapeutic use of retinoids against gastric cancer.

• 한국응용과학기술학회지
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• 제30권1호
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• pp.173-182
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• 2013

아토피성 피부염은 만성 재발성 염증성 피부질환으로 피부장벽기능의 이상과 환경유발인자에 대한 피부과민성과 연관되어 있다. 이전의 연구에서는 아토피성피부염에 효과적인 약용식물 추출물을 발굴하기 위하여 노회, 자회지정, 석류, 석곡 추출물들의 세포독성, 항산화, 항염, 항알레르기 효과를 검토하였다. 본 연구에서는 지질다당류로 활성화시킨 대식세포 RAW26.7 에 대한 약용식물 추출물들의 항염작용을 보다 상세하게 검토하여 항염작용의 근본적인 분자기전을 확인하고자 하였다. 역전사중합효소연쇄반응분석(reverse transcription polymerase chain reaction analysis) 결과, 석류, 석곡, 노회는 염증성 사이토카인인 IL-6 와 IL-$1{\beta}$ 유전자발현을 현저하게 억제시켰으며 자화지정은 영향이 없었다. 형질주입과 발광효소분석(transfection and luciferase analysis) 결과, 약용식물 모두가 전사 핵인자 카파비(NF-${\kappa}B$)의 활성화를 억제시켰다. 웨스턴 블럿 분석(western blot analysis) 결과, 노회는 JNK MAP 인산화효소의 활성화를 차단하였지만 p38 MAP 인산화효소의 활성화는 차단하지 못하였다. 반면에 자화지정, 석류, 석곡은 JNK MAP 인산화효소뿐만 아니라 p38 MAP 인산화효소의 활성화도 차단하였다. 이들 실험결과들은 노회, 자화지정, 석류, 석곡은 항염효능을 가지고 있으며 따라서 아토피성 피부염의 증상을 경감 또는 완화시키는 잠재력이 있음을 보여 준다.

• 한국식품영양과학회지
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• 제44권12호
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• pp.1912-1917
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• 2015

천연식물에 광범위하게 존재하는 대표적인 페닐프로파노이드 화합물인 caffeic acid에 대해 배 유래의 polyphenol oxidase로 산화반응을 수행하여 상대적으로 높은 pancreatic lipase 저해 활성($IC_{50}$; $161.2{\pm}2.8{\mu}g/mL$)을 확인하였으며, 이는 caffeic acid와 비교하였을 경우 활성이 상승함을 알 수 있었다. Caffeic acid 산화반응물에 대해서 $C_{18}$ 겔을 활용한 column chromatography를 수행하여 4종의 리그난 화합물을 분리하였고, 각 화합물의 화학구조는 NMR 스펙트럼 데이터 해석 및 표품과의 HPLC 직접 비교를 통하여 phellinsin A(2), caffeicinic acid(3), isocaffeicinic acid(4), 7,8-erythro-caffeicin(5)으로 동정하였다. 이들 화합물중 phellinsin A(2)는 $IC_{50}$ 값이 $66.3{\pm}2.6{\mu}M$로 가장 강한 효능을 나타내었으며, 다음으로 caffeic acid 2분자의 산화 결합을 통해 생합성된 caffeicinic acid(3)의 $IC_{50}$ 값이 $109.6{\pm}3.7{\mu}M$의 저해능을 나타내었다. 배에 존재하는 polyphenol 산화효소에 의해 생합성된 caffeic acid 이량체가 pancreatic lipase 저해 활성 물질임을 확인하였으며, 이들 활성은 caffeic acid가 결합 양상에 따른 화합물의 구조에 따라 다름이 시사되었다. 향후 이들 활성물질의 활성 기작에 대한 연구가 필요하며 본 연구 결과는 보다 우수한 pancreatic lipase 저해능을 가지는 새로운 선도화합물 발굴을 위한 기초자료로 이용될 수 있을 뿐만 아니라 항비만 물질의 상업화를 위한 기초자료로 이용될 수 있을 것으로 사료된다.

• 한국식품영양과학회지
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• 제45권8호
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• pp.1208-1213
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• 2016

천연물로부터 당뇨합병증에 효과적인 소재를 개발하기 위하여 본 연구를 수행하였으며, 울릉도산 마가목 열매의 80% methyl alcohol 추출물의 ethyl acetate 가용부로부터 최종 당화산물 생성 억제능($IC_{50}$; $181.2{\pm}2.8{\mu}g/mL$)을 확인하였다. 효능성분의 동정을 위하여 $C_{18}$ 겔 등을 활용한 column chromatography를 수행하여 3종의 페놀성 화합물을 분리하였고, 각 화합물의 화학구조는 NMR 스펙트럼 데이터 해석 및 표품과의 HPLC 직접 비교를 통하여 neochlorogenic acid(1), cryptochlorogenic acid(2) 및 chlorogenic acid (3)로 동정하였다. 이들 화합물에 대해 최종당화산물 생성억제능을 평가한 결과 neochlorogenic acid(1)가 가장 강한 $167.5{\pm}3.5{\mu}M$의 $IC_{50}$ 값을 나타내었고, cryptochlorogenic acid(2)는 $185.9{\pm}3.7{\mu}M$의 $IC_{50}$ 값을 나타내었다. 또한, 이들 물질에 대해 $ONOO^-$ 소거 활성을 평가한 결과 cryptochlorogenic acid(2)가 가장 강한 라디칼 소거 활성인 $4.0{\pm}0.2{\mu}M$의 $IC_{50}$ 값을 나타내었고, 구조이성질체인 neochlorogenic acid(1)가 $4.5{\pm}0.3{\mu}M$의 $IC_{50}$ 값을 나타내었으며, 이들 활성은 caffeic acid가 결합 양상에 따른 화합물의 구조에 따라 다름이 시사되었다. 향후 이들 활성 물질의 활성 기작에 대한 연구가 필요하며 본 연구 결과는 더욱 우수한 최종당화산물 생성 억제능을 가지는 새로운 선도화합물 발굴을 위한 기초자료로 이용될 수 있을 뿐만 아니라 당뇨합병증 예방 및 치료에 효과적인 천연물질의 상업화를 위한 기초자료로 이용될 수 있을 것으로 생각한다.

• 한국식품영양과학회지
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• 제41권8호
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• pp.1057-1065
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• 2012

본 연구는 생약의 휘발성 향기추출액을 이용한 염증치료용 기능성 신소재를 개발하기 위해 대식세포주인 Raw 264.7 세포를 이용한 모델시스템에서 천궁 및 당귀의 휘발성 향기추출물의 항염증 효과를 검정하였으며, 아울러 이들 향기추출물의 향기성분을 GC-MS 분석을 통해 확인하였다. 그 결과 8가지 생약(노간주, 목향, 천궁, 당귀, 정향, 고본, 박하, 애엽)의 향기추출물 중 천궁 및 당귀가 LPS 처리에 의한 NO의 생성을 가장 크게 억제하였으며 $PGE_2$의 생성도 현저히 억제시키는 것으로 나타났다. 또한, 천궁 및 당귀의 향기 추출물은 NO의 생합성 효소인 iNOS 단백질의 발현 또한 농도의존적으로 감소시킴을 확인할 수 있었으며, 염증성 사이토카인인 IL-6의 생성 억제효과도 우수하였다. 천궁 및 당귀 향기추출물의 휘발성 향기성분을 GC-MS로 분석한 결과, 천궁은 eugenol, 3-butylphthalide, butylidene phthalide 및 ligustilide를 확인하였으며 이들 향기성분 중 ligustilide가 천궁의 주된 휘발성 향기성분임을 확인하였고, 당귀는 eugenol, ${\beta}$-maaliene, ${\beta}$-eudesmol 및 9-tetradecenoic acid를 확인하였으며 이들 향기성분 중 ${\beta}$-eudesmol이 당귀의 주된 향기성분임을 확인하였다. 이러한 연구결과로 미루어 볼 때 천궁 및 당귀의 휘발성 향기추출물이 대식세포에 의해 생성되는 염증반응의 매개물질인 NO, $PGE_2$, IL-6의 생성을 저해함으로써 항염증 활성을 가지는 것으로 생각되며, 이러한 항염증 활성은 천궁 및 당귀에 함유된 항염증성 향기성분인 ligustilide 및 ${\beta}$-eudesmol이 크게 기여했을 것으로 사료된다. 따라서 천궁 및 당귀의 휘발성 향기추출물은 천연 유래의 항염증 물질로서 잠재적인 가치가 있어 향후 염증질환 예방 및 치료용 기능성식품 및 화장품 소재로 활용할 수 있을 것으로 생각된다.

• Asian-Australasian Journal of Animal Sciences
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• 제33권1호
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• pp.127-131
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• 2020

Objective: This study was conducted to determine catalytic properties of wheat phytase with exopolyphosphatase activity toward medium-chain and long-chain inorganic polyphosphate (polyP) substrates for comparative purpose. Methods: Exopolyphosphatase assay of wheat phytase toward polyP75 (medium-chain polyP with average 75 phosphate residues) and polyP1150 (long-chain polyP with average 1150 phosphate residues) was performed at pH 5.2 and pH 7.5. Its activity toward these substrates was investigated in the presence of Mg²⁺, Ni²⁺, Co²⁺, Mn²⁺, or ethylenediaminetetraacetic acid (EDTA). Michaelis constant (K_m) and maximum reaction velocity (V_max) were determined from Lineweaver-Burk plot with polyP75 or polyP1150. Monophosphate esterase activity toward p-nitrophenyl phosphate (pNPP) was assayed in the presence of polyP75 or polyP1150. Results: Wheat phytase dephosphorylated polyP75 and polyP1150 at pH 7.5 more effectively than that at pH 5.2. Its exopolyphosphatase activity toward polyP75 at pH 5.2 was 1.4-fold higher than that toward polyP1150 whereas its activity toward polyP75 at pH 7.5 was 1.4-fold lower than that toward polyP1150. Regarding enzyme kinetics, K_m for polyP75 was 1.4-fold lower than that for polyP1150 while V_max for polyP1150 was 2-fold higher than that for polyP75. The presence of Mg²⁺, Ni²⁺, Co²⁺, Mn²⁺, or EDTA (1 or 5 mM) exhibited no inhibitory effect on its activity toward polyP75. Its activity toward polyP1150 was inhibited by 1 mM of Ni²⁺ or Co²⁺ and 5 mM of Ni²⁺, Co²⁺, or Mg²⁺. Ni²⁺ inhibited its activity toward polyP1150 the most strongly among tested additives. Both polyP75 and polyP1150 inhibited the monophosphate esterase activity of wheat phytase toward pNPP in a dose-dependent manner. Conclusion: Wheat phytase with an unexpected exopolyphosphatase activity has potential as a therapeutic tool and a next-generational feed additive for controlling long-chain polyP-induced inappropriate inflammation from Campylobacter jejuni and Salmonella typhimurium infection in public health and animal husbandry.

• Asian Pacific Journal of Cancer Prevention
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• 제14권9호
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• pp.5199-5205
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• 2013

Background/Aim: The Hippo signaling pathway is a newly discovered and conserved signaling cascade, which regulates organ size control by governing cell proliferation and apoptosis. This study aimed to investigate its effects in human gastric cancer. Methods: Tumor tissues (n=60), adjacent non-tumor tissues (n=60) and normal tissues (n=60) were obtained from the same patients with primary gastric cancer (GC). In addition, 70 samples of chronic atrophic gastritis (CAG) tissues were obtained from patients with intestinal metaplasia (IM) by endoscopic biopsy. Hippo signaling molecules, including Mst1, Lats1, YAP1, TAZ, TEAD1, Oct4 and CDX2, were determined by quantitative polymerase chain reaction (qPCR). Protein expression of Mst1, Lats1, YAP1, TEAD1 and CDX2 was assessed by immunohistochemistry and Western blotting. Results: Mst1, Lats1 and Oct4 mRNA expression showed an increasing tendency from GC tissues to normal gastric tissues, while the mRNA expression of YAP1, TAZ and TEAD1 was up-regulated (all P<0.01). Mst1 and Lats1 protein expression presented a similar trend with their mRNA expression. In addition, YAP1 and TEAD1 protein expression in GC was significantly higher than in the other groups (all P<0.01). CDX2 mRNA and protein expression in the CAG group were higher than in the other groups (all P<0.01). In GC, mRNA expression of Mst1, Lats1, Oct4, YAP1, TAZ, TEAD1 and CDX2 had a close correlation with lymphatic metastasis and tumor TNM stage (all P<0.01). Furthermore, protein expression of Mst1, Lats1, YAP1, TAZ, TEAD1 and CDX2 had a close correlation between each other (P<0.05). Conclusion: The Hippo signaling pathway is involved in the development, progression and metastasis of human gastric cancer. Therefore, manipulation of Hippo signaling molecules may be a potential therapeutic strategy for gastric cancer.

• Journal of Acupuncture Research
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• 제34권2호
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• pp.1-18
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• 2017

Objectives : The purpose of this study was to evaluate the effects of water extracts of Eucommiae cortex (EC), Psoraleae semen (PS), and their combination on receptor activator of nuclear factor-kappa-B ligand (RANKL)-induced osteoclast differentiation. Methods : We assayed the protein expression levels of nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1), c-Fos, mitogen-activated protein kinases (MAPKs), and ${\beta}-actin$ in cell lysates using western blotting. Similarly, mRNA expression levels of NFATc1, c-Fos, tartrateresistant acid phosphate (TRAP), and glyceraldehyde-3-phosphate dehydrogenase, spermatogeni (GAPDHS) from bone marrow macrophages (BMMs) were analyzed using reverse transcription-polymerase chain reaction (RT-PCR). Furthermore, we determined the anti-osteoporotic effects of the water extracts of EC, PS, and their combination in a lipopolysaccharide (LPS)-induced bone-loss mouse model. Results : The in vitro data revealed showed that the combination of EC and PS extract showed a more remarkable inhibition of osteoclast differentiation than each herb did alone. The combination downregulated the induction of c-Fos, NFATc1, and TRAP by suppressing the phosphorylation of p38 and c-Jun N-terminal kinases (JNKs) and inhibiting nuclear factor kappa-light-chain-enhancer of activated B cells ($NF-{\kappa}B$). Lastly, the in vivo data showed that PS reduced the LPS-induced bone erosion. Conclusion : The result of this study suggests that EC and PS could be potential therapeutic agents for bone loss diseases such as osteoporosis.