Juglone inhibits epithelial-mesenchymal transition of prostate cancer cells by regulating Wnt/β-catenin/Snail signaling pathway

AIM: To investigate the mechanism of juglone on epithelial-mesenchymal transition in prostate cancer cells. METHODS: Human prostate cancer LNCaP cells were divided into control group (without juglone), 12.5 μmol/L juglone group and 25 μmol/L juglone group. LNCaP cells in the latter 2 groups were treated with juglone for 24 h. The invasion ability of the LNCaP cells was detected by Transwell assay. The protein expression of E-cadherin, vimentin, Snail and β-catenin was determined by Western blot. The LNCaP cells were treated with LiCl and juglone in combination for 24 h, and the protein expression of Snail and E-cadherin was detected by Western blot.RESULTS: The results of Trans-well invasion assay showed that the invasion ability in juglone groups was significantly decreased (P<0.01). The protein expression of E-cadherin in the LNCaP cells treated with juglone was increased, and the expression levels of vimentin and β-catenin were reduced (P<0.01). Treatment with LiCl significantly attenuated the inhibitory effect of juglone on Snail expression and subsequent down-regulation of E-cadherin expression. CONCLUSION: Juglone inhibits the epithelial-mesenchymal transition by inhibiting the Wnt/β-catenin/Snail signaling pathway in the LNCaP cells.     

上皮间质转化促乳腺癌机制及药物干预治疗研究进展

乳腺癌是犬、猫等伴侣动物与人类常发疾病,作为人类及动物常患恶性肿瘤和主要致死肿瘤之一,其疾病负担仍呈逐步加重趋势,乳腺癌的预防及治疗形势愈加严峻。上皮间质转化(EMT)是乳腺癌发生发展中重要的生物学过程。EMT还可促进恶性肿瘤的侵袭、扩散及耐药,因此它在肿瘤的研究中日益受到关注,靶向于EMT是治疗乳腺癌的重要研究方向与热点。文章就EMT发生过程中细胞形态功能及标志物的变化、EMT分类及EMT与乳腺癌的关系分别展开论述,详细解析了EMT相关TGF-β/Smad、NF-κB及Wnt信号通路转导途径;随后对乳腺癌治疗药物研究进展,包括TGF-β/Smad通路抑制剂开发,相关药物、基因及细胞因子治疗前景、NF-κB通路与Wnt通路抑制剂的动物试验研究结果进行了详细论述;最后对乳腺癌的治疗发展与趋势进行了展望。深入认识信号通路调控乳腺癌EMT的生物学过程,明确其发生发展机制,寻找关键靶点及开发靶向药物,将为乳腺癌的精准治疗带来曙光。     

Inhibitory effect of AuNPs-Endostar on melanoma lung metastasis in mice

AIM: To observe the influence of gold nanoparticles combined with Endostar (AuNPs-Endostar) on the melanoma lung metastasis of mice and the underlying mechanism. METHODS: C57BL/6 mice (n=24) were selected for constructing the model of spontaneous lung metastasis of melanoma B16-F10 cells. Subsequently, the mice were randomly divided into Endostar group, AuNPs group, AuNPs-Endostar group and model group. After the formation of melanoma, the mice in each group were injected with different drugs through tail vein for 0.1 mL daily. After 9 d, the mice were narcotized for cutting the tumors in situ. After the operation, they were raised for 2 weeks before killed for obtaining the lung tissues to observe the situation of the metastasis. HE staining was utilized for observing the necrosis status of the tumors in situ, while immunostaining was applied for testing the expression of CD31, carbonic anhydrase-IX (CA-IX), vimentin and zonula occludens-1 (ZO-1) in the tumors. RESULTS: Compared with model group, the pulmonary metastasis in the groups with medical treatment was obviously reduced. In AuNPs-Endostar group, the metastasis inhibition rate was the highest, and the tumor necrosis was also decreased obviously, with the significant reduction of CD31, CA-IX and vimentin expression in the tumors and significant increase in ZO-1 expression. CONCLUSION: Compared with using Endostar or AuNPs alone, the combination of AuNPs with Endostar significantly improves the curative effect of inhibiting the pulmonary metastasis of melanoma in the mice. The mechanism may be related to reducing the tumor angiogenesis, norma-lizing the blood vessels and improving tumor hypoxia, thus inhibiting the tumor epithelial-mesenchymal transition, increasing the tight junctions between tumor cells and decreasing the invasiveness.     

Low potassium diets with different levels of calcium in comparison with different anionic diets fed to prepartum dairy cows: Effects on sorting behaviour,total tract digestibility,energy metabolism,oxidative status and hormonal response

This study investigated the effects of low potassium diets with different levels of Ca compared to two diets low in dietary cation–anion difference (DCAD) fed prepartum as a strategy to prevent hypocalcemia on sorting behaviour, total tract digestibility, oxidative status and energy and protein metabolism of transition cows. Forty-eight pregnant dairy cows were assigned to 4 treatment groups: Low Ca, low K (LCLK), High Ca, low K (HCLK), Supplementation with anionic mineral mixture (AMS) supplementation with SoyChlor (CAS). After parturition, all animals were fed a standard postpartum diet. Data were collected until 21 DIM. Prepartum urinary pH was significantly reduced by the low DCAD diets, while postpartum Ca homeostasis was affected by the HCLK ration. Feeding AMS induced sorting against particles <1.18 mm in favour of particles >19 mm prepartum. In contrast, cows fed CAS showed an increase in selective consumption of fine particles and sorted against longer particles similar to the HCLK and LCLK groups. Postpartum sorting activity was not affected by the dietary treatments. After calving, apparent digestibility of NDF was significantly reduced in the HCLK group. Prepartum, we observed effects on serum concentrations of non-esterified fatty acids were higher and insulin sensitivity was lower in the AMS group. Blood urea nitrogen (BUN) was decreased in cows fed the CAS ration. Postpartum, we found serum protein to be decreased with the low DCAD diets while BUN was decreased in the CAS group. The low DCAD rations increased prepartum serum malondialdehyde concentrations, while postpartum total antioxidant capacity was lower in the HCLK and the AMS group. From these data, we conclude that AMS decreased prepartum intake due to compromised palatability. Intermediate protein metabolism was affected by the low DCAD diets, while parameters of oxidative stress were probably affected by acid–base balance and Ca homeostasis.     

半干旱农牧交错区土壤养分空间变异特征分析——以内蒙古伊金霍洛旗为例

本文以伊金霍洛旗为研究区,通过GIS平台对土壤养分中的有机质、全氮、速效钾的空间插值方法进行了比较并得到了最优插值图。然后分析了该旗土壤养分的空间分布规律,为该地区的农业生产布局和生态环境建设提供了依据。通过比较后认为,在该地区有机质的空间插值用反距离权重法为好,而全氮和速效钾的空间插值以克吕格法的指数模型和高斯模型为好。地统计学分析结果表明:有机质、全氮和速效钾的变程分别为11853m,8932.2m和732.99m,全氮和速效钾具有中等空间相关性,而有机质空间相关性较弱。有机质含量普遍偏低,在91%的面积中有机质含量低于10g/kg,其相对高值区呈离散分布,可能与隐域植被的分布有关。全氮含量与有机质含量分布相似,在70%的面积中低于0.75g/kg,而速效钾含量中等,介于102.04-103.31mg/kg之间。在三个不同的景观亚区中,乌审凹地亚区的土壤养分状况较好,黄土丘陵亚区养分状况最差,毛乌素沙地亚区介于它们之间。     

Modelling land cover transitions: A solution to the problem of spatial dependence in data

Raster-based spatial land cover transition models (LCTMs) are widely used in landscape ecology. However, many LCTMs do not account for spatial dependence of the input data, which may artificially fragment the output spatial configuration. We demonstrate the consequences of ignoring spatial dependence, thus assigning probabilities randomly in space, using a simple LCTM. We ran the model from four different initial conditions with distinct spatial configurations and results indicated that, after 20 simulation steps, all of them converged towards the spatial configuration of the random data set. From an ecological perspective this is a serious problem because ecological data often exhibit distinct spatial configuration related to ecological processes. As a solution, we propose an approach (region approach) that accounts for spatial dependence of LCTM input data. Underlying spatial dependence was used to apply spatial bias to probability assignment within the model. As a case study we applied a region approach to a Vegetation Transition Model (VTM); a semi-Markovian model that simulates forest succession. The VTM was applied to approximately 500,000 ha of boreal forest in Ontario, at 1 ha pixel resolution. When the stochastic transition algorithms were applied without accounting for spatial dependence, spatial configuration of the output data became progressively more fragmented. When the VTM was applied using the region approach to account for spatial dependence output fragmentation was reduced. Accounting for spatial dependence in transition models will create more reliable output for analyzing spatial patterns and relating those patterns to ecological processes.This revised version was published online in May 2005 with corrections to the Cover Date.     

非瞬时或突变的马尔可夫链的一种应用方法

介绍对于一种非瞬时或突变的状态转移过程.采用Bayes方法修订马尔可夫过程的状态转移概率矩阵;将先验概率分布修订为后验概率分布;对整个过程的信息加以吸收,得出能够反映整个过程的状态转移概率矩阵,从而达到预测目的.由此还得出一个重要结论:吸收信息的无序性.     

Markov积分半群的逼近

通过证明Markov积分Q半群的逼近证明了l∞上一个Markov积分半群G(t)可以由一族一致收敛的积分半群逼近.     

Calreticulin promotes migration and invasion of nasopharyngeal carcinoma cells by inducing cell EMT

AIM:To observe the expression of calreticulin (CRT) in nasopharyngeal carcinoma tissues, analyze the significance of clinical pathology and the influence on epithelial-mesencymal transition (EMT) of CNE2 cells. METHODS:The expression of calreticulin was detected by immunohistochemistry in 52 nasopharyngeal carcinoma and 57 nasopharyngeal benign tissues, and the significance of clinical pathology was evaluated. The calreticulin gene-specific small interfering RNA was constructed, and then was transfected into the NPC cell line CNE2 using the cationic liposome method. The effect of CRT on the morphological changes of the CNE2 cells was observed under light microscope. The effect of CRT on the cell migration and invasion abilities of the CNE2 cells was detected by Transwell migration and invasion assays. The expression of EMT-related proteins E-cadherin, vimentin, transforming growth factor (TGF)-β and matrix metalloproteinase (MMP)-9 in the CNE2 cells was determined by Western blot. RESULTS:The positive expression rate of CRT in the benign lesion tissues was 19.29% (11/57), which was significantly increased in the nasopharyngeal carcinoma tissues as 82.69% (43/52). The expression rate of CRT was positively correlated with the stage of nasopharyngeal carcinoma and lymph node metastasis (P<0.05). Knockdown of CRT expression made the CNE2 cells showing a spindle shape to a flat, cobblestone-like epithelial state change, arranged more compact, and the migration and invasion abilities were significantly decreased (P<0.05). Knockdown of CRT expression resulted in significant increase in the protein expression of E-cadhe-rin, and the decreases in the protein expression of vimentin, TGF-β and MMP-9 in the CNE2 cells (P<0.05). CONCLUSION:Calreticulin expression in nasopharyngeal carcinoma is significantly higher and positively correlated with nasopharyngeal carcinoma stage and lymph node metastasis. Calreticulin promotes cell migration and invasion of nasopharyngeal carcinoma CNE2 cells by inducing EMT.     

Role of ILK siRNA on expression of GSK-3β and β-catenin in human tubular epithelial cells stimulated by high glucose

AIM：To investigate the effects of siRNA targeting integrin-linked kinase (ILK) on the expression of glycogen synthase kinase 3β (GSK-3β) and β-catenin during epithelial-mesenchymal transition (EMT) in human kidney proximal tubular epithelial cell line HKC induced by high glucose. METHODS：HKC cells were divided into 4 groups:normal glucose (NG) group, high glucose (HG) group, HG+HK (a vector containing the non-specific siRNA designed as negative control) group and HG＋ILK siRNA group. The inverted fluorescence microscope was used to examine the expression of green fluorescent protein (GFP). The expression of ILK at mRNA and protein levels was detected by RT-PCR and Western blotting. The expression of p-GSK-3β and β-catenin was observed by immunocytochemical staining. The protein expression of total GSK-3β, p-GSK-3β, nuclear β-catenin, total β-catenin, E-cadherin and α-smooth muscle actin (α-SMA) was measured by Western blotting. RESULTS：GFP was observed in HKC cells, indicating that the transfection was successful. Both the protein and mRNA of ILK were down-regulated in HG＋ILK siRNA group compared with HG group and HG+HK group, but still higher than those in NG group. Silencing of ILK down-regulated the expression of p-GSK-3β and nuclear β-catenin. No difference of total GSK-3β or total β-catenin was observed among the 4 groups. CONCLUSION:These data support a functional role of ILK, GSK-3β and β-catenin in tubular EMT induced by high glucose. ILK may promote tubular EMT by regulating the activity of GSK-3β and β-catenin, the downstream effectors of the Wnt/β-catenin pathway.