• Asian Pacific Journal of Cancer Prevention
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• 제13권11호
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• pp.5455-5461
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• 2012

Luteolin is a plant flavonoid which exhibits anti-oxidative, anti-inflammatory and anti-tumor effects. However, the antiproliferative potential of luteolin is not fully understood. In this study, we investigated the effect of luteolin on cell cycling and apoptosis in human esophageal squamous carcinoma cell line Eca109 cells. MTT assays showed that luteolin had obvious cytotoxicity on Eca109 with an $IC_{50}$ of $70.7{\pm}1.72{\mu}M$ at 24h. Luteolin arrested cell cycle progression in the G0/G1 phase and prevented entry into S phase in a dose- and time-dependent manner. as assessed by FCM. Luteolin induced apoptosis of Eca109 cells was demonstrated by AO/EB staining assay and annexin V-FITC/PI staining. Moreover, luteolin downregulated the expression of cyclin D1, survivin and c-myc, and it also upregulated the expression of p53, in line with the fact that luteolin was able to inhibit Eca109 cell proliferation.

• Asian Pacific Journal of Cancer Prevention
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• 제15권4호
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• pp.1531-1537
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• 2014

Runt-related transcription factor 3 (RUNX3) is a tumor suppressor gene whose reduced expression may play an important role in the development and progression of esophageal squamous cell cancer (ESCC). The aim of this study was to investigate the clinical relevance of RUNX3 in ESCC patients and effects of overexpression on biological behaviour of Eca109 cells in vitro and in vivo. Immunohistochemistry was performed to detect the clinical relevance of RUNX3 and lymph node metastasis in 80 ESCC tissues and 40 non-cancerous tissues using the SP method. RT-PCR and Western blotting were applied to assess the RUNX3 level and verify the Eca109 cell line with stable overexpression. Localization of RUNX3 proteins was performed by cell immunofluorescence. CCK-8 and Scrape motility assays were used to determine proliferation and migration and the TUNEL assay to analyze cell apoptosis. Invasive potential was assessed in cell transwell invasion experiments. In nude mice, tumorigenesis in vivo was determined. Results showed decreased expression of RUNX3 in esophageal tissue to be significantly related to lymph node metastasis (LNM) (P<0.01). In addition, construction of a recombinant lentiviral vector and transfection into the human ESCC cell line Eca109 demonstrated that overexpression could inhibit cell proliferation, migration and invasion, and induce apoptosis. The in vivo experiments in mice showed tumorigenicity and invasiveness to be significantly reduced. Taken together, our studies indicate that underexpression of RUNX3 in human ESCC tissue is significantly correlated with progression. Restoration of RUNX3 expression significantly inhibits ESCC cells proliferation, migration, invasion and tumorigenesis.

• Asian Pacific Journal of Cancer Prevention
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• 제13권1호
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• pp.205-209
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• 2012

Background: The inhibition of tumor cell growth without toxicity to normal cells is an important target in cancer therapy. One possible way to increase the efficacy of anticancer drugs and to decrease toxicity or side effects is to develop traditional natural products, especially from medicinal plants. Paris polyphylla Smith has shown anti-tumour effects by inhibition of tumor promotion and inducement of tumor cell apoptosis, but mechanisms are still not well understood. The present study was to explore the effect of Paris polyphylla Smith extract (PPSE) on connexin26 and growth control in human esophageal cancer ECA109 cells. Methods: The effects of PPSE on Connexin26 were examined by RT-PCR, western blot and immunofluorescence; cell growth and proliferation were examined by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) assay. Results: PPSE inhibited the growth and proliferation on esophageal cancer ECA109 cells, while increasing the expression of connexin26 mRNA and protein; conversely, PPSE decreased Bcl-2 and increased Bad. Conclusion: This study firstly shows that PPSE can increase connexin26 expression at mRNA and protein level, exerting anti-tumour effects on esophageal cacner ECA109 cells via inhibiting cell proliferation and inducing cell apoptosis.

• Asian Pacific Journal of Cancer Prevention
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• 제13권9호
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• pp.4531-4536
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• 2012

Objective: To study synergistic effects of nedaplatin and cisplatin on three human carcinoma cell lines (esophageal carcinoma cell line Eca-109, ovarian carcinoma Skov-3 and cervical carcinoma Hela). Methods: Inhibition effects were evaluated by MTT assay and cell apoptosis was detected by flow cytometry. In addition, changes of Ki-67, Bax and Bcl-2 at mRNA and protein levels were quantified by RT-PCR and Western blotting. Results: Growth inhibition in each cell lines was dose-dependent after exposure to nedaplatin or cisplatin alone. The interaction of the two drugs was synergistic at higher concentrations according to the median-effect principle. The inhibition rates with nedaplatin, cisplatin and combined treatment were $41.9{\pm}4.1%$, $47.4{\pm}2.9%$, $52.5{\pm}0.9%$(Eca-109), $39.0{\pm}1.26%$, $45.0{\pm}1.45%$, $56.2{\pm}1.44%$ (Skov-3) and $44.8{\pm}2.11%$, $46.9{\pm}0.99%$, $56.6{\pm}1.83%$ (Hela) respectively, with increase in apoptosis. Compared with the nedaplatin or cisplatin alone treatment group, the combinative treatment group's Ki-67 and bcl-2 mRNA (protein) expression was decreased while that of Bax mRNA (protein) was increased. Conclusion: Compared to the effects of nedaplatin or cisplatin alone at high concentrations, combination of nedaplatin and cisplatin at low concentrations proved to be much more effective for inhibition of proliferation and the induction of apoptosis in the Eca-109, Skov-3 and Hela cell lines.

• 한국토양비료학회지
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• 제39권2호
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• pp.109-115
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• 2006

점토경반층 토양에서 수량 변이의 특성을 파악하기 위하여 센서를 통하여 관측된 깊이별 토양경도 및 ECa와 작물의 수량과의 관련성을 분석한 결과, 깊이별 토양의 경도는 점토가 집적된 지표면으로부터 15-30 cm 지점의 점토경반층 (argillic horizon)에서 높게 나타내고 있으며 토양의 깊이가 깊은 지점에서 토양경도의 변이가 작게 나타났다. 1994년부터 2002년까지 콩과 옥수수 수량과 작물생육기의 강우량을 분석한 결과 7-8월의 강우량이 작물의 수량과 매우 밀접한 관계를 가지고 있으며 이 기간의 강우량이 150 mm 이하이면 작물이 수분 부족으로 수량이 낮아지는 것으로 나타났다. 지표로부터 15-45 cm 지점에서 토양의 경도와 ECa 가 작물의 수량과 매우 유의한 상관을 갖는 것으로 나타났으며 "drought boundary" 인 7-8월의 강우량이 150 mm를 기점으로 각기 반대의 상관을 보이는 것으로 나타났다. 측정된 토양의 깊이별 경도 값과 ECa 를 이용하여 수량 추정식을 계산 하였으며 추정식의 검증을 위하여 별도의 독립적인 자료를 이용하여 추정된 수량과 측정된 수량의 표준오차를 비교한 결과 측정된 수량에 대한 표준오차의 비율이 4-16% 로 나타났으며 7-8월의 강우량이 150 mm 이하로 건조한 경우에 표준오차가 같은 시기에 강우량이 150 mm 이상으로 습윤한 경우보다 현저하게 표준오차가 크게 나타났다. 결론적으로 신속하고 경제적으로 이용할 수 있는 센서 측정자료와 작물수량과의 관련성을 분석한 결과, 연구에 이용된 점토경반층 토양에서 센서를 이용하여 측정한 ECa 및 깊이별 토양경도와 작물 수량간에 통계적으로 유의한 상관이 있음을 알 수 있다.

• Asian Pacific Journal of Cancer Prevention
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• 제13권6호
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• pp.2527-2532
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• 2012

Objectives: The aim of the present study was to explore mechanisms underlying the effects of down-regulating ${\beta}$-catenin expression on esophageal carcinoma (EC) cells. Methods: Cell cycle distribution and apoptosis were determined using flow cytometry and annexin V apoptosis assay, respectively. Transmission electron microscopy (TEM) was used to examine changes in ultrastructure, while expression of cyclin D1 protein and mRNA was detected by western blot and real-time PCR. Proliferating cell nuclear antigen (PCNA) and extracellular signal-regulated kinase (ERK) 1-2 were evaluated by Western blot analysis. PCNA labeling index (LI) was determined by immunocytochemistry. Results: Compared with pGen-3-con transfected and Eca-109 cells, the percentage of G0/G1-phase pGen-3-CTNNB1 transfected cells was obviously increased (P<0.05), with no significant difference among the three groups with regard to apoptosis (P>0.05). pGen-3-CTNNB1 transfected cells exhibited obvious decrease in cyclin D1 mRNA and protein expression (P<0.05) and the ultrastructure of Eca-109 cells underwent a significant change after being transfected with pGen-3-CTNNB1, suggesting that down-regulating ${\beta}$-catenin expression can promote the differentiation and maturation. The expression of PCNA and the ERKI/2 phosphorylation state were also down-regulated in pGen-3-CTNNB1 transfected cells (P<0.05). At the same time, the PCNA labeling index was decreased accordingly (P<0.05). Conclusion: Inhibition of EC Eca-109 cellproliferation by down-regulating ${\beta}$-catenin expression could improve cell ultrastructure by mediating blockade in G0/G1 through inhibiting cyclin D1, PCNA and the MAPK pathway (p-ERK1/2).

• Asian Pacific Journal of Cancer Prevention
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• 제15권20호
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• pp.8679-8684
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• 2014

CD133 was recently reported to be a cancer stem cell and prognostic marker. Quercetin is considered as a potential chemopreventive agent due to its involvement in suppression of oxidative stress, proliferation and metastasis. In this study, the expression of CD133/CD44 in esophageal carcinomas and Eca109/9706 cells was explored. In immunoflurorescence the locations of $CD133^+$ and multidrug resistance 1 $(MDR1)^+$ in the same E-cancer cells were coincident, mainly in cytomembranes. In esophageal squamous cell carcinomas detected by double/single immunocytochemistry, small $CD133^+$ cells were located in the basal layer of stratified squamous epithelium, determined as CSLC (cancer stem like cells); $CD44^+$ surrounding the cells appeared in diffuse pattern, and the larger $CD44^+$ (hi) cells were mainly located in the prickle cell layer of the epithelium, as progenitor cells. In E-cancer cells exposed to nanoliposomal quercetin (nLQ with cytomembrane permeability), down-regulation of NF-${\kappa}Bp65$, histone deacetylase 1 (HDAC1) and cyclin D1 and up-regulation of caspase-3 were shown by immunoblotting, and attenuated HDAC1 with nuclear translocation and promoted E-cadherin expression were demonstrated by immunocytochemistry. In particular, enhanced E-cadherin expression reflected the reversed epithelial mesenchymal transition (EMT) capacity of nLQ, acting as cancer attenuator/preventive agent. nLQ acting as an HDAC inhibitor induced apoptotic cells detected by TUNEL assay mediated via HDAC-NF-${\kappa}B$ signaling. Apoptotic effects of liposomal quercetin (LQ, with cytomembrane-philia) combined with CD133 antiserum were also detected by CD133 immunocytochemistry combined with TUNEL assay. The combination could induce greater apoptotic effects than nLQ induced alone, suggesting a novel anti-CSC treatment strategy.

• 한국분무공학회지
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• 제16권2호
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• pp.104-109
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• 2011

The present study investigated experimentally the spreading characteristics of a single liquid impinging on the inclined micro-textured aluminum (Al 6061) surfaces manufactured by using a micro computerized numerical control (${\mu}$-CNC) milling machine. The textured surfaces were composed of patterned micro-holes (diameter of $125\;{\mu}m$ and depth of $125\;{\mu}m$). In our experiment, the de-ionized (DI) water droplet of $4.3\;{\mu}l$ was impinged normally on the non-textured and textured surfaces at two different Weber numbers, and the droplet impinged on the inclined surfaces with different angles. A high speed camera was used to capture sequential digital images for measurement of the maximum spreading distance. It was found that for the textured surface, the measured apparent equilibrium contact angle (ECA) increased up to $105.8^{\circ}$, higher than the measured ECA of $87.6^{\circ}$ for the non-textured (bare) surface. In addition, it is conjectured that the spreading distance decreased because of a liquid penetration during droplet spreading through the holes, the increase in hydrophobicity, and viscous dissipation during impact process.

• Asian Pacific Journal of Cancer Prevention
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• 제14권9호
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• pp.5391-5396
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• 2013

Aim: To investigate the role of Golgi phosphoprotein 3 (GOLPH3) in tumour growth and metastasis of esophageal squamous cancer. Methods: A lentiviral shRNA-vector was utilized to stably knockdown GOLPH3 in Eca-109 esophageal squamous cancer cells. mRNA transcription and protein expression of GOLPH3 were examined by real-time quantitative PCR and Western blotting, respectively. Cell proliferation activity was assessed by MTT assay and invasion and migration potentials by matrigel invasion and transwell motility assays. Results: Stable knockdown in the GOLPH3 cell line was established. PD-A gene expression was significantly suppressed by lentivirus-mediated RNAi, which resulted in reducing the capacity for cell proliferation, migration, invasion and adhesion in vitro. In vivo, GOLPH3 depletion resulted in inhibition of tumour growth, with stable decrease in the expression of GOLPH3 in tumor xenografts. Conclusions: Our findings suggest that lentivirus mediated silencing of the GOLPH3 gene has a significant anti-tumour effect on esophageal squamous cancer in vitro and in vivo. In addition, the results indicate that GOLPH3 might be an effective molecular target for gene therapy in esophageal squamous cancer.

• Asian Pacific Journal of Cancer Prevention
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• 제15권23호
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• pp.10299-10306
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• 2015

The esophageal squamous cell carcinoma (ESCC) is thought to develop through a multi-stage process. Epigenetic gene silencing constitutes an alternative or complementary mechanism to mutational events in tumorigenesis. Posttranscriptional regulation of human leukocyte antigen class I (HLA-I) and antigen processing machinery (APM) proteins expression may be associated with novel epigenetic modifications in cancer development. In the present study, we determined the expression levels of HLA-I antigen and APM components by immunohistochemistry. Then by a bisulfite-sequencing PCR (BSP) approach, we identified target CpG islands methylated at the gene promoter region of APM family genes in a ESCC cell line (ECa109), and further quantitative analysis of CpG site specific methylation of these genes in cases of Kazakh primary ESCCs with corresponding non-cancerous esophageal tissues using the Sequenom MassARRAY platform. Here we showed that the development of ESCCs was accompanied by partial or total loss of protein expression of HLA-B, TAP2, LMP7, tapasin and ERp57. The results demonstrated that although no statistical significance was found of global target CpG fragment methylation level sof HLA-B, TAP2, tapasin and ERp57 genes between ESCC and corresponding non-cancerous esophageal tissues, there was significant differences in the methylation level of several single sites between the two groups. Of thesse only the global methylation level of LMP7 gene target fragments was statistically higher ($0.0517{\pm}0.0357$) in Kazakh esophageal cancer than in neighboring normal tissues ($0.0380{\pm}0.0214$, p<0.05). Our results suggest that multiple CpG sites, but not methylation of every site leads to down regulation or deletion of gene expression. Only some of them result in genetic transcription, and silencing of HLA-B, ERp57, and LMP7 expression through hypermethylation of the promoters or other mechanisms may contribute to mechanisms of tumor escape from immune surveillance in Kazakh esophageal carcinogenesis.