Interaction between uranium and humic acid (Ⅱ): complexation, precipitation and migration behavior of U(Ⅵ) in the presence of humic substances

The complexation, precipitation, and migration behavior of uranium in the presence of humic acid (HA) or fulvic acid (FA) were investigated by cation exchange, ultrafiltration and dynamic experiment, respectively. The results showed that (i) complex equilibrium between the uranium and humic substances was achieved at approximately 72 h, (ii) the coordination number varied from 1:1 to 1:2 ( U(Ⅵ) : humic acid) as pH increased from 3 to 6; and (iii) , while the complex stability constant decreased when temperature increased, but increased with pH value. We found that the precipitation of uranyl could only be observed in presence of HA, and the precipitation was influenced by conditions, such as pH, uranium concentration, temperature, and the HA concentration. The maximum precipitation proportion up to 60% could be achieved in the condition of 40 mg/L HA solution at pH 6. We further observed that the migration behavior of uranium in soil in the presence of humic acid (HA) or fulvic acid (FA) was different from that in the presence of inorganic colloid, and the effect of humic substances (HS) was limited.

Activation of adult endogenous neurogenesis by a hyaluronic acid collagen gel containing basic fibroblast growth factor promotes remodeling and functional recovery of the injured cerebral cortex

The presence of endogenous neural stem/progenitor cells in the adult mammalian brain suggests that the central nervous system can be repaired and regenerated after injury.However,whether it is possible to stimulate neurogenesis and reconstruct cortical layers II to VI in non-neurogenic regions,such as the cortex,remains unknown.In this study,we implanted a hyaluronic acid collagen gel loaded with basic fibroblast growth factor into the motor cortex immediately following traumatic injury.Our findings reveal that this gel effectively stimulated the proliferation and migration of endogenous neural stem/progenitor cells,as well as their differentiation into mature and functionally integrated neurons.Importantly,these new neurons reconstructed the architecture of cortical layers II to VI,integrated into the existing neural circuitry,and ultimately led to improved brain function.These findings offer novel insight into potential clinical treatments for traumatic cerebral cortex injuries.

Effects of rAAV-CD151 and rAAV-antiCD151 on the Migration of Human Tongue Squamous Carcinoma Cell Line Tca8113

Summary: This study was designed to determine the effects of the recombinant adeno-associated virus vector containing sense CD151 gene (rAAV-CD151) and antisense CD151 gene (rAAV-antiCD151) on the migration of Tca8113 cell. Functional fragment of CD151 gene was amplified by RT-PCR, and inserted into the vector pAAV in the sense direction and antisense direction, respectively. The rAAV-CD151 and rAAV-antiCD151 were produced and the titers were determined by dot blot. The CD151, at protein level, was detected by Western blot. The Transwell chamber was used to detect the effects of the rAAV-CD151 and rAAV-antiCD151 on the tumor cell migration. The titers of the rAAV-CD151 and rAAV-antiCD151 were 2×10~11 pfu/ml and 1.0×10~11 pfu/ml, respectively. The expression of CD151 was increased by 108 % in the cells transfected with rAAV-CD151 and decreased by 79 % in the cells transfected with rAAV-antiCD151, as compared with non-transfected cells, respectively. The number of the migrating cells was significantly increased in the cells transfected with rAAV-CD151 (93.56±11.59) and decreased in the cells transfected with rAAV-antiCD151 (24.00±4.36) as compared with non-transfected and rAAV-GFP transfected cells (53.00±6.56 and 46.00±7.00, P<0.05). It is an important molecular mechanism of the tumor metastasis that the overexpression of CD151 promotes the migration of the tumor cells. The rAAV-antiCD151 is a novel tool, which can reduce the expression of CD151 and inhibit the migration of the tumor cells, and brings us a new approach of anti-sene gene therapy targeted at CD151 in human carcinoma.

Effects of microtubule-associated protein tau expression on neural stem cell migration after spinal cord injury

Our preliminary proteomics analysis suggested that expression of microtubule-associated protein tau is elevated in the spinal cord after injury. Therefore, the first aim of the present study was to examine tau expression in the injured spinal cord. The second aim was to determine whether tau can regulate neural stem cell migration, a critical factor in the successful treatment of spinal cord injury. We established rat models of spinal cord injury and injected them with mouse hippocampal neural stem cells through the tail vein. We used immunohistochemistry to show that the expression of tau protein and the number of migrated neural stem cells were markedly increased in the injured spinal cord. Furthermore, using a Transwell assay, we showed that neural stem cell migration was not affected by an elevated tau concentration in the outer chamber, but it was decreased by changes in intracellular tau phosphorylation state. These results demonstrate that neural stem cells have targeted migration capability at the site of injury, and that although tau is not a chemokine for targeted migration of neural stem cells, intracellular tau phosphorylation/dephosphorylation can inhibit cell migration.

Suppression of Human Liver Cancer Cell Migration and Invasion via the GABA_A Receptor

Objective To investigate the roles of the y-aminobutyric acid （GABA） in the metastasis of hepatocellular carcinoma （HCC） and to explore the potential of a novel therapeutic approach for the treatment of HCC. Methods The expression levels of GABA receptor subunit genes in various HCC cell lines and patients＇ tissues were detected by quantitative real-time polymerase chain reaction and Western blot analysis. Transwell cell migration and invasion assays were carried out for functional analysis. The effects of GABA on liver cancer cell cytoskeletal were determined by immunofluorescence staining. And the effects of GABA on HCC metastasis in nude mice were evaluated using an in vivo orthotopic model of liver cancer. Results The mRNA level of GABA receptor subunits varied between the primary hepatocellular carcinoma tissue and the adjacent non-tumor liver tissue. GABA inhibited human liver cancer cell migration and invasion via the ionotropic GABAA receptor as a result of the induction of liver cancer cell cytoskeletal reorganization. Pretreatment with GABA also significantly reduced intrahepatic liver metastasis and primary tumor formation in vivo. Conclusions These findings introduce a potential and novel therapeutic approach for the treatment of cancer patients based on the modulation of the GABAergic system.

Relationship between entero-hepatic bile acid circulation and interdigestive migrating myoelectrical activity in rats

AIM： To investigate the effects of entero-hepatic bile add circulation on the inter-digestive migrating myoelectrical complex （MMC） in rats. METHODS： Thirty-two rats were divided into four groups. Three pairs of bipolar silver electrodes were chronically implanted in the antrum, duodenum and jejunum. Three groups of Uhem were ligatecl around the upper part of common bile duct （CBD）. The experiments were performed in consdous and fasting state. The gastrointestinal myoelectrical activity was recorded. Ursodeoxycholic acid （UDCA） and saline were then perfused into stomachs of two groups with CBD obstruction and the effects of them on the MMC were observed. RESULTS： A typical pattern of MMC was observed in normal fasting rats. MMC of antral and duodenal origin disappeared temporarily in earlier stage of CBD obstruction. While MMC of jejunum origin appeared. increased MMC cycle duration was seen after 4 d in rats with CBD obstruction. The MMC after CBD obstruction was characterized by an increased duration of phase Ⅱ-like activity and decreased duration of phase Ⅰ ＆ Ⅲ activity. Perfusion into stomachs with UDCA resulted in a shorter MMC cycle duration and a longer duration of phase III of duodenal origin compared to the normal group. CONCLUSION： Entero-hepatic bile add drculation initiates inter-digestive MMC of duodenal origin.

Anti-tumor activity of rice bran hydrolysates on migration, invasion and angiogenesis

Objective:To investigate anti-tumor effect of rice bran hydrolysates(RBH)on proliferation,migration,invasion,and angiogenesis of cholangiocarcinoma(CCA)cells,and elucidate the underlying mechanisms.Methods:RBH was prepared from Tubtim Chumprae rice(Oryza sativa L.)by hydrothermolysis followed by protease digestion.Phenolic content in RBH was analyzed by high-performance liquid chromatography.Human CCA cells,KKU-156,KKU-452,and KKU-100,were used to study the effects of RBH on proliferation,migration,invasion,and adhesion by wound healing,Transwell chamber,and fibronectin cell adhesion assays.Angiogenesis was evaluated using human umbilical vein endothelial cells.Proteins associated with cancer progression were analyzed by immunobloting assays.Results:RBH contained carbohydrates,proteins,lipids,and various phenolic compounds and flavonoids.RBH did not inhibit CCA proliferation,but strongly suppressed migration,invasion,adhesion of CCA cells,and the formation of tube-like capillary structures of human umbilical vein endothelial cells.Moreover,RBH downregulated phosphorylation of FAK,PI3K,and Akt,suppressed NF-κB nuclear translocation,decreased the expression of ICAM-1,vimentin and vascular endothelium growth factor(VEGF),and increased the expression of E-cadherin.Conclusions:RBH suppresses CCA cell migration and invasion and decreases expression of proteins involved in cancer metastasis.RBH is a potential food supplement for cancer prevention.

茯苓酸对小鼠黑色素瘤细胞增殖、迁移和侵袭的影响及机制研究

目的:研究茯苓酸(pachymic acid,PA)对小鼠黑色素瘤B16细胞增殖、迁移和侵袭的影响及其可能机制。方法:采用CCK8法和细胞克隆形成实验检测不同浓度(20、40、80μmol/L)PA对B16细胞增殖的影响;细胞划痕和Transwell小室实验观察PA对B16细胞迁移和侵袭能力的影响;Western Blot法和免疫荧光法检测PA对B16细胞的上皮-间质转化相关蛋白表达的变化。结果:PA可以明显抑制B16细胞增殖,并呈一定的浓度依赖性关系(P<0.01);PA干预24 h后可以显著抑制B16细胞的迁移和侵袭能力(P<0.01);40μmol/L的PA能上调B16细胞的E-cadherin蛋白表达(P<0.05)并下调Vimentin蛋白的表达(P<0.05);免疫荧光法检测显示40μmol/L的PA可明显降低B16细胞Vimentin蛋白的表达。结论:PA可以抑制B16细胞的增殖、迁移和侵袭,其机制可能与上调B16细胞的E-cadherin蛋白表达,降低Vimentin蛋白表达,从而抑制B16细胞的上皮-间质转化有关。

基于稻谷加温加湿技术的γ-氨基丁酸富化

目的:提高糙米品质。方法:以γ-氨基丁酸(GABA)含量和谷氨酸脱羧酶活性作为指标,研究富化条件对GABA含量的影响;基于最优参数,通过分析富化过程中籽粒各部位GABA含量,揭示其迁移规律;最后分析该法对糙米品质的影响,并与浸泡法对比。结果:原料水分含量14.5%,富化温度65℃,最终水分含量20%,富化时间5 h时,GABA含量最高。富化过程中,GABA主要在糠层生成,在生成的同时向内部迁移,各碾减率的糙米GABA含量均在5 h达到峰值,但迁移率在4 h后趋于稳定,胚乳部分的GABA迁移率最高可达87.1%。与浸泡法相比,加温加湿法富化的糙米偏黄,爆腰率较低,米饭更柔软。结论:加温加湿法优于浸泡法,可用于制备高GABA含量的糙米和精米。

Gabra3通过AKT/mTOR途径促进膀胱癌T24细胞侵袭和迁移

目的:探究γ-氨基丁酸受体亚基α-3(Gamma-aminobutyric acid receptor subunit alpha-3,Gabra3)基因对膀胱癌T24细胞凋亡、迁移和侵袭的作用及其机制。方法:TCGA数据库分析Gabra3基因在膀胱癌中的表达以及转移和生存的相关性。建立Gabra3过表达和Gabra3突变的T24细胞株,根据转染质粒不同,分为空白T24细胞(Control)、空pDONR223载体转染T24细胞(NC)、野生型Gabra3过表达T24细胞株(wt-Gabra3)、突变型Gabra3 T24细胞株(mut-Gabra3)。在回补实验中,分为转染突变型Gabra3 T24组(mut-Gabra3)、转染空pDONR223载体mut-Gabra3组(mut-Gabra3-NC)、转染野生型Gabra3过表达组(mut-Gabra3+wt-Gabra3)。用TUNEL染色检测T24细胞凋亡,细胞划痕和Transwell分别检测T24细胞迁移和侵袭,Western blot检测AKT/mTOR通路相关分子生物表达水平。结果:Gabra3基因在膀胱癌中显著高表达(P<0.05),生存曲线证实远处转移存在的患者生存期明显较短(P<0.05)。成功构建Gabra3过表达和突变的T24细胞株。与Control组相比,wt-Gabra3组细胞凋亡能力显著降低,迁移、侵袭能力显著增强(P<0.05),而mut-Gabra3组细胞凋亡显著增加,侵袭能力显著减弱(P<0.05);wt-Gabra3组细胞p-AKT、p-mTOR、p-4E-BP1、p-S6K蛋白表达均显示上凋(P<0.05),而mut-Gabra3组细胞上述蛋白无差异。在回补实验中,过表达wt-Gabra3的mut-Gabra3突变T24细胞凋亡能力受到抑制(P<0.05),迁移和侵袭能力显著增强(P<0.05),并且该组细胞AKT/mTOR通路相关分子显著激活(P<0.05)。结论:外源性的未编辑的Gabra3可以促进膀胱癌T24细胞的迁移、侵袭能力,并且可能与激活AKT/mTOR途径通路密切相关。

miR-409-3p通过靶向脂肪酸结合蛋白4影响子宫内膜癌细胞转移的研究

目的用生物信息学方法预测脂肪酸结合蛋白4(FABP4)的上游调控miRNA,体外培养子宫内膜癌细胞Ishikawa,了解微小核糖核酸-409-3p(miR-409-3p)是否通过抑制FABP4的表达水平,进而影响Ishikawa细胞的生物学行为。方法实验分为control组、miR-409-3p mimics组、miR-409-3p inhibitor组、miR-409-3p NC组、FABP4-OE组及FABP4-shRNA组6组,采用实时定量聚合酶链式反应及蛋白质印迹法检测miR-409-3p及FABP4在Ishikawa中的表达情况;采用细胞增殖实验(CCK-8)、细胞侵袭及迁移实验(Transwell法)检测miR-409-3p与FABP4对Ishikawa细胞增殖、侵袭以及迁移能力的影响;用双荧光素酶报告基因实验在Ishikawa细胞中检测miR-409-3p对FABP4的调控作用。结果1)在Ishikawa细胞中下调FABP4的表达后,Ishikawa细胞的增殖、侵袭和迁移能力均显著降低(P<0.05);在Ishikawa细胞中上调FABP4的表达后,Ishikawa细胞的增殖、侵袭和迁移能力均显著升高(P<0.05)。2)在Ishikawa细胞中转染miR-409-3p mimics后,FABP4的表达水平以及Ishikawa细胞的增殖、侵袭和迁移能力均显著降低(P<0.05);而转染miR-409-3p inhibitor后,FABP4的表达水平以及Ishikawa细胞的增殖、侵袭和迁移能力均显著升高(P<0.05)。3)双荧光素酶报告基因结果显示,在Ishikawa细胞中miR-409-3p可靶向调控FABP4。结论高表达的FABP4促进Ishikawa细胞增殖、侵袭及迁移,miR-409-3p可通过下调FABP4的表达进而抑制Ishikawa细胞发生转移。

没食子酸对人骨肉瘤HOS细胞增殖、迁移和侵袭能力及上皮-间充质转化的影响

目的探讨没食子酸(GA)对人骨肉瘤HOS细胞增殖、迁移和侵袭能力以及上皮-间充质转化(EMT)的影响。方法CCK-8试剂盒检测没食子酸对人骨肉瘤HOS细胞存活率的影响。细胞划痕愈合实验和Transwell小室检测没食子酸对HOS细胞迁移和侵袭能力的影响。Western blot检测HOS细胞E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)和Snail蛋白表达水平。结果没食子酸能显著抑制人骨肉瘤HOS细胞的增殖、迁移与侵袭;没食子酸显著上调E-cadherin蛋白表达,下调N-cadherin、Vimentin、Snail蛋白表达。结论没食子酸可以抑制人骨肉瘤HOS细胞的增殖、迁移、侵袭及EMT。

腐植酸水溶肥部分替代化学氮肥对土壤-水稻体系镉迁移富集的影响

为探明腐植酸水溶肥部分替代化学氮肥对调控镉(Cd)在土壤-水稻体系吸收转运效果,采用盆栽试验方法,设置不施氮肥(CK)、化学氮肥(CF)、替代20%化肥氮(HF1)、替代45%化肥氮(HF2)及替代70%化肥氮(HF3)5个处理,探究腐植酸水溶肥部分替代化学氮肥对Cd高污染酸性稻田土壤有效态Cd含量及水稻各部位Cd吸收、转运和积累的影响。结果表明:施氮和腐植酸水溶肥替代氮肥处理均不同程度降低土壤pH、增加土壤有效态Cd含量,土壤有效态Cd含量随腐植酸水溶肥替代比例增加而升高,但差异不显著;施氮和腐植酸水溶肥替代氮肥处理促进水稻植株各部位对Cd的吸收富集,显著影响器官间Cd的转运。与CF处理相比,HF2和HF3处理分别显著提高Cd转运系数TF_(叶片/茎秆)90.00%和190.00%,分别提高叶片Cd富集系数100.00%和233.33%,分别降低转运系数TF_(糙米/穗轴)35.06%和41.6%,抑制了Cd向稻米迁移分配;施氮能有效促进干物质形成,与CF处理相比,HF2处理显著提高糙米和地上部干物质重,分别达26.82%和23.03%,有利于稻米干物质重形成。本试验条件下,腐植酸水溶肥适宜替代氮肥45%~70%能提高水稻干物质重,有效降低Cd向稻米迁移富集。

碱性条件下Cr(Ⅵ)和腐殖酸对赤铁矿胶体在多孔介质中迁移过程的机理研究

赤铁矿胶体是一种广泛存在于土壤中的活性组分,可与重金属Cr(Ⅵ)发生吸附、沉淀、络合等多种化学作用,从而影响Cr(Ⅵ)在土壤中的迁移富集.探究赤铁矿胶体-Cr(Ⅵ)-腐殖酸(HA)三元体系在多孔介质中的共迁移过程,对于深入理解Cr(Ⅵ)污染过程和防控原理具有重要意义.该研究通过吸附、聚集和柱迁移实验,探究了碱性条件下Cr(Ⅵ)和HA对赤铁矿胶体的聚集动力学和迁移能力的影响.结果表明:与不加Cr(Ⅵ)相比,加入0.25~10 mg/L Cr(Ⅵ)会提高赤铁矿胶体的ζ电势,降低胶体与石英砂之间的最大排斥势垒,阻滞赤铁矿胶体在砂柱中迁移,穿透峰值从61%降至50%左右.引入HA会增加赤铁矿胶体表面负电荷,增强Cr(Ⅵ)与赤铁矿胶体之间的静电斥力,且与Cr(Ⅵ)竞争吸附赤铁矿胶体表面的结合位点,抑制Cr(Ⅵ)在赤铁矿胶体上的吸附,导致胶体与石英砂之间的最大排斥势垒升高.Cr(Ⅵ)和HA的吸附先后顺序对赤铁矿胶体稳定性和迁移性有显著影响.赤铁矿胶体在HA-Cr(Ⅵ)体系中的ζ电势和水合粒径(分别为−30 mV和200~300 nm)比Cr(Ⅵ)-HA体系(分别为−28 mV和400~500 nm)要小,且在HA-Cr(Ⅵ)体系中穿透峰值更高.研究显示,在碱性条件下,尽管HA会抑制Cr(Ⅵ)在赤铁矿胶体表面上的吸附,但会增加赤铁矿胶体在环境中的稳定性和迁移性,使得吸附在赤铁矿胶体表面的Cr(Ⅵ)随胶体发生共迁移的可能性增加,具有潜在的环境风险.

miR-21对人口腔癌细胞株HSQ-89增殖、侵袭与迁移的影响实验研究

目的:探讨微小核糖核酸-21(miR-21)介导Smad7调节转化生长因子β(TGF-β)/Smad信号通路对人口腔癌细胞株HSQ-89增殖、侵袭与迁移的作用。方法:取人口腔癌细胞株HSQ-89培养传代,分为阴性对照组、下调组、上调组和正常组,前三者分别采用脂质体转染法将携带阴性对照(NC inhibitor)、miR-21抑制剂(miR-21 inhibitor)、miR-21模拟物(miR-21 mimics)载体进行转染,正常组仅添加等量无菌蒸馏水。观察各组细胞增殖、侵袭、迁移能力;实时-定量聚合酶链反应(RT-qPCR)检测各组细胞miR-21、TGF-β_(1)、Smad3、Smad7、细胞周期蛋白D1(CCND1)、MYC、E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)、基质金属蛋白酶-2(MMP-2)mRNA表达;蛋白免疫印迹法(WB)检测各组细胞TGF-β_(1)、Smad3、Smad7、CCND1、MYC、E-cadherin、Vimentin、MMP-2蛋白表达及p-Smad3水平;双荧光素酶报告基因检测验证miR-21是否靶向Smad7。结果:与正常组和阴性对照组比较,上调组细胞增殖、侵袭、迁移能力上升,下调组细胞增殖活性下降、侵袭细胞数减少、划痕愈合率下降(均P<0.05);与正常组和阴性对照组比较,上调组细胞miR-21表达、TGF-β_(1)、Smad3、CCND1、MYC、Vimentin、MMP-2 mRNA和蛋白表达上升、Smad7和E-cadherin mRNA和表达下降,差异有统计学意义(均P<0.05),下调组细胞miR-21表达、TGF-β_(1)、Smad3、CCND1、MYC、Vimentin、MMP-2 mRNA和蛋白表达下降、Smad7和E-cadherin mRNA和蛋白表达上升(均P<0.05);经双荧光素酶报告基因实验验证miR-21靶向Smad7。结论:miR-21促进口腔癌细胞增殖、侵袭和迁移,可能与调节Smad7、TGF-β/Smad信号通路相关。

SH3BGRL对胃癌细胞增殖、迁移的影响

目的探讨SH3结构域结合富含谷氨酸的蛋白质(SH3 binding glutamic acid-rich protein like,SH3BGRL)对胃癌细胞增殖、迁移能力的影响及其作用机制。方法基于TCGA、GTEx及GEO数据库,通过生物信息学分析SH3BGRL在胃癌组织及正常胃黏膜上皮组织中的表达情况;通过HPA数据库获取SH3BGRL蛋白在胃癌组织及正常胃黏膜上皮组织中的表达图谱;并基于TCGA数据库的SH3BGRL表达水平进行高、低表达分组,分析其对胃癌患者预后、免疫细胞浸润和免疫治疗的影响。通过临床标本IHC染色观察SH3BGRL蛋白在胃癌中的表达情况;采用RT-qPCR检测SH3BGRL在胃癌细胞系和正常胃黏膜上皮细胞GES-1中的表达水平。将胃癌HGC-27及NCI-N87细胞进行siRNA转染分为si-NC组(转染阴性对照组)、si-SH3BGRL组(沉默SH3BGRL组),并通过RT-qPCR及Western blot验证是否转染成功。采用CCK-8及Transwell实验检测细胞增殖和迁移能力,随后诱导THP-1单核细胞分化为M0巨噬细胞,并将分化后的M0巨噬细胞和转染后的胃癌细胞进行共培养,采用RT-qPCR及细胞免疫荧光检测共培养之后M2巨噬细胞标志物的改变,并采用RT-qPCR检测SH3BGRL沉默胃癌细胞中CSF1的表达水平。结果生物信息学分析结果显示,与正常胃黏膜上皮组织相比,SH3BGRL在胃癌组织中表达上调(均P<0.01),且SH3BGRL高表达者总生存期和无进展生存期比SH3BGRL低表达的患者短(均P<0.01)。SH3BGRL高表达者M2巨噬细胞浸润比例更高,与免疫治疗低应答率相关(均P<0.01)。IHC染色结果表明,SH3BGRL蛋白在胃癌组织中表达上调(P<0.001)。与GES-1细胞相比,SH3BGRL在胃癌HGC-27及NCI-N87细胞中的mRNA表达水平均显著上调(均P<0.01)。与si-NC组相比,沉默SH3BGRL后HGC-27和NCI-N87细胞的增殖能力明显下降(均P<0.01),HGC-27细胞迁移能力显著下降(P<0.01)。M0巨噬细胞与沉默SH3BGRL的HGC-27及NCI-N87共培养后,M2巨噬细胞标志物CD163、CD206及Arg1均下调(均P<0.01)。在HGC-27及NCIN87细胞中沉默SH3BGRL后,NF-κB信号通路的靶基因CSF1显著下调(均P<0.01)。结论SH3BGRL在胃癌中表达上调,沉默SH3BGRL能抑制胃癌细胞增殖和迁移,其机制可能与抑制M2巨噬细胞极化进而形成免疫抑制肿瘤微环境有关。

CD147通过Akt/mTOR信号通路调控脂肪酸合成对子宫颈癌细胞增殖、侵袭和迁移的影响

目的探讨CD147的表达对子宫颈癌细胞增殖、侵袭、迁移的影响和潜在的分子机制。方法分析UCSC数据库BSG基因(编码CD147蛋白)在子宫颈癌中的表达,采用Log-rank test法评估各组表达的预后差异。应用Western blot法检测CD147在Siha、Hela和H8细胞中的表达,通过慢病毒转染Hela细胞下调CD147表达,并验证其转染效率。运用Western blot法检测各组细胞中p-Akt、p-mTOR、ACC1、FASN、E-cadherin和N-cadherin的表达;使用BODIPY染色和脂肪酸试剂盒检测细胞中脂肪酸含量;利用CCK-8、平板克隆和Transwell实验检测细胞增殖、侵袭和迁移能力。sh-CD147组经Akt激动剂(SC79)处理后,Western blot法检测细胞中p-Akt、p-mTOR、ACC1、FASN、E-cadherin和N-cadherin的表达;选用BODIPY染色和脂肪酸试剂盒检测细胞中脂肪酸含量;利用平板克隆实验和Transwell实验分别检测细胞增殖、侵袭和迁移能力。结果UCSC数据库显示CD147在子宫颈癌中的表达高于正常子宫颈组织(P<0.01);CD147过表达的患者预后不良。Western blot检测结果显示,与H8细胞相比,Siha和Hela细胞中CD147蛋白表达增高(P=0.011)。下调CD147表达后,与Hela组相比,sh-CD147组细胞中CD147、ACC1和FASN蛋白表达降低(P<0.001);BODIPY荧光染色减弱,脂肪酸含量下降(P<0.001);细胞集落形成能力、侵袭和迁移能力降低;sh-CD147组细胞中E-cadherin蛋白表达升高,N-cadherin、p-Akt和p-mTOR表达降低。与sh-CD147相比,经Akt激动剂SC-79剂处理后(sh-CD147-SC79)细胞内p-Akt、p-mTOR、ACC1、FASN和N-cadherin表达升高,E-cadherin表达降低,脂质染色和脂肪酸含量检测结果与关键酶表达一致(P<0.01),细胞增殖、侵袭迁移能力显著增强。结论CD147通过Akt/mTOR信号通路调控脂肪酸合成,促进子宫颈癌细胞增殖、侵袭和迁移。

白酒厂污泥与生活污泥有机组分的差异性及其淋溶迁移风险评估

通过对白酒厂污泥和生活污泥中腐植酸、低分子有机酸的含量对比分析,探讨两种污泥中有机组分的差异性及其淋溶迁移的环境风险。结果表明,酒厂污泥中有机质和腐植酸(NaOH浸提)平均含量分别为428.2,342.47 g/kg,平均比生活污泥中的含量高47.7%,62.6%。从污泥浸提(H 3PO 4浸提)的低分子有机酸含量,白酒厂污泥中柠檬酸和苹果酸含量分别达3.90,0.88 g/kg,分别是生活污泥的10.00倍和1.83倍;而生活污泥中的草酸含量达3.74 g/kg,是白酒厂污泥的2.94倍。两种污泥的腐植酸均含有丰富的羟基、酚羟基,而白酒厂污泥的腐植酸还含有丰富的羧基。两种污泥的6次连续淋溶的实验表明,生活污泥的淋溶液中,COD、NH_(4)^(+)-N含量范围分别为121.5~380.5,1.83~10.67 mg/L,平均比酒厂污泥高出55.8%,27.2%;而白酒厂污泥的淋溶液中,TP含量范围为0.84~1.90 mg/L,平均比生活污泥淋溶液中的含量高28.1%,白酒厂污泥有机组分淋溶迁移对水环境的影响小于生活污泥。综上所述,白酒厂污泥的有机组分及腐植酸的资源化利用优于生活污泥。

小檗碱对人胶质瘤T98G细胞迁移和侵袭的抑制作用及其机制

目的:探讨小檗碱(BBR)对人胶质瘤T98G细胞脂肪酸的调控作用及细胞增殖、迁移和侵袭的影响,阐明其潜在的作用机制。方法:对数生长期T98G细胞分为对照组和不同浓度(25、50及100 mg·L^(-1))BBR组,采用细胞划痕愈合实验检测各组细胞迁移率,Transwell小室实验检测各组细胞侵袭率。对数生长期T98G细胞分为对照组和100 mg·L^(-1)BBR组,质谱法检测2组细胞中脂肪酸含量。对数生长期T98G细胞分为对照组和不同浓度(50、100及150 mg·L^(-1))BBR组,采用Western blotting法检测各组细胞中磷脂酰肌醇3-激酶(PI3K)、磷酸化PI3K(p-PI3K)、蛋白激酶B(AKT)、磷酸化AKT(p-AKT)、固醇调节元件结合蛋白1(SREBP-1)和脂肪酸合成酶(FASN)蛋白表达水平。应用基因沉默技术抑制FASN表达,将对数生长期T98G细胞分为对照组、shFASN1组和shFASN2组,采用Western blotting法检测各组细胞中FASN蛋白表达水平,克隆形成实验检测各组细胞克隆形成情况,细胞划痕愈合实验检测各组细胞迁移率。结果:与对照组比较,不同浓度BBR组细胞迁移率和侵袭率呈浓度依赖性降低(P<0.01),100 mg·L^(-1)BBR组细胞中各类脂肪酸含量明显降低(P<0.01)。与对照组比较,150 mg·L^(-1)BBR组细胞中p-PI3K、p-AKT、SREBP-1和FASN蛋白表达水平均明显降低(P<0.05或P<0.01),100和150 mg·L^(-1)BBR组细胞中SREBP-1蛋白表达水平明显降低(P<0.01)。抑制FASN表达后,与对照组比较,shFASN1组和shFASN2组细胞中FASN蛋白表达水平均明显降低(P<0.01),且shFASN2组细胞中FASN蛋白表达水平低于shFASN1组(P<0.05);与对照组比较,shFASN1组和shFASN2组细胞中克隆形成数明显减少,细胞迁移率明显降低(P<0.01),且shFASN2组细胞迁移率明显低于shFASN1组(P<0.05)。结论:BBR通过降低细胞中PI3K/AKT/SREBP-1/FASN通路相关蛋白的表达干扰胶质瘤T98G细胞中脂肪酸合成,进而降低其迁移和侵袭能力。

单宁酸抑制膀胱癌细胞增殖及迁移侵袭作用的实验研究

目的:探讨中药五倍子提取物单宁酸对膀胱癌EJ细胞和T24细胞的增殖、迁移和侵袭作用及其机制。方法:采用CCK-8、细胞生长计数实验方法检测不同浓度的单宁酸对膀胱癌EJ细胞和T24细胞增殖的抑制作用,应用平板克隆、细胞划痕和Transwell实验方法检测单宁酸对EJ和T24细胞增殖、迁移及侵袭能力的影响,Western Blot实验检测单宁酸对膀胱癌EJ和T24细胞凋亡、转移及维甲酸相关孤儿核受体C蛋白(RORC)的表达作用。结果:不同浓度的单宁酸对膀胱癌EJ细胞、T24细胞增殖的抑制作用呈剂量依赖性(P<0.05),EJ和T24细胞增殖、迁移、侵袭能力随着单宁酸浓度增加而降低(P<0.05);单宁酸上调膀胱癌EJ和T24细胞的凋亡相关蛋白PARP1、Caspase7以及E-cadherin的表达;同时下调转移相关蛋白N-cadherin、MMP-2、Vimentin、SNAI1的表达;EJ和T24细胞凋亡相关蛋白的表达与RORC的上调相关。结论:单宁酸通过上调RORC表达抑制膀胱癌EJ、T24细胞的增殖及迁移侵袭作用。