Tunable Colloidal Properties of Lauramidopropyl Betaine and Li Co-modified Montmorillonite in Ethanol/Water.

Montmorillonite (Mt) is a hydrophilic clay mineral with a generally high cationic exchange capacity and a remarkable swellability in water. Yet the application of Mt in cosmetics, paints, polymer nanocomposites, drug delivery systems, and tissue engineering are limited due to its unfavorable swelling and dispersion in alcohol/water mixtures. Improving the swellability and dispersibility of Mt in mixtures of ethanol and water remains challenging. Here, we showed that the swellability and dispersibility of Mt in ethanol/water could be significantly enhanced when lithium-Mt (Li-Mt) was intercalated by zwitterionic surfactant lauramidopropyl betaine (LPB). The binding mechanism of the LPB intercalate to Li-Mt originated from a combination of van der Waals forces, ion-dipole interaction, and electrostatic attraction. Due to the synergistic effect of Li+ and LPB, the comodified Mt (LPB-Li-Mt) exhibited excellent swellability, dispersibility, and rheological properties. The structure, morphology, zeta potential, dispersibility, and gel-forming performance of LPB-Li-Mt can be modulated by the concentrations of ethanol in ethanol/water mixtures. When the ethanol concentration increased to 75% v/v ethanol solution, the free swelling of LPB-Li-Mt remained above 80%. The results from X-ray diffraction, thermogravimetric analysis, Fourier transform infrared spectroscopy, X-ray photoemission spectrometry, and small-angle X-ray scattering confirmed the full exfoliation of LPB-Li-Mt at 75% (v/v) ethanol solution. The formation of a stable colloidal LPB-Li-Mt dispersion in a mixture of ethanol/water might be derived from the association between water molecules and the Li+, the hydrophobic interaction, and the ion-dipole of ethanol with the LPB molecules. The findings provide a guide for improving dispersion and swelling of Mt and modified ones in water-miscible organic solvents.

Controlled fabrication of Ag@clay nanomaterials for ultrasensitive and rapid surface-enhanced Raman spectroscopic detection.

The nanostructure of Ag nanoparticles (NPs) plays a critical role in their surface-enhanced Raman scattering (SERS) activity. Despite many efforts to tune the nanostructure of Ag NPs, it remains a great challenge as Ag NPs tend to agglomerate and their nanostructure is difficult to control. Herein, newly-discovered clay-surfactant-Ag+ materials and interfacial processes were developed and used to prepare uniform spherical Ag@synthetic hectorite (Ag@Hct) nanomaterials for ultrasensitive SERS assay. Sodium dodecyl sulfate (SDS), an anionic surfactant, acted as a bridge to conjugate the positively charged edge of Hct NPs and Ag+via electrostatic interaction to form the bridging nanostructure of Hct-SDS-Ag+, which promoted the uniform dispersion of Hct NPs. Following this, Ag+ was reduced to Ag0 by the reductant, and Ag0 grew on the surface of disc-like Hct NPs to form spherical Ag@Hct nanomaterials with an average particle size of ∼24 nm. The prepared Ag@Hct nanomaterials showed an ultrasensitive SERS response to methylene blue (MB) with a detection limit of 10-12 M. The detection limit of MB in sewage was 10-11 M. The prepared Ag@Hct nanomaterials also exhibited great SERS enhancement for malachite green and crystal violet. This work provides a novel and simple approach to prepare Ag@Hct nanomaterials with uniform spheres and adjustable particle size, allowing more sensitive and reproducible detection of MB.

Calcium carbonate: controlled synthesis, surface functionalization, and nanostructured materials.

Calcium carbonate (CaCO3) is an important inorganic mineral in biological and geological systems. Traditionally, it is widely used in plastics, papermaking, ink, building materials, textiles, cosmetics, and food. Over the last decade, there has been rapid development in the controlled synthesis and surface modification of CaCO3, the stabilization of amorphous CaCO3 (ACC), and CaCO3-based nanostructured materials. In this review, the controlled synthesis of CaCO3 is first examined, including Ca2+-CO32- systems, solid-liquid-gas carbonation, water-in-oil reverse emulsions, and biomineralization. Advancing insights into the nucleation and crystallization of CaCO3 have led to the development of efficient routes towards the controlled synthesis of CaCO3 with specific sizes, morphologies, and polymorphs. Recently-developed surface modification methods of CaCO3 include organic and inorganic modifications, as well as intensified surface reactions. The resultant CaCO3 can then be further engineered via template-induced biomineralization and layer-by-layer assembly into porous, hollow, or core-shell organic-inorganic nanocomposites. The introduction of CaCO3 into nanostructured materials has led to a significant improvement in the mechanical, optical, magnetic, and catalytic properties of such materials, with the resultant CaCO3-based nanostructured materials showing great potential for use in biomaterials and biomedicine, environmental remediation, and energy production and storage. The influences that the preparation conditions and additives have on ACC preparation and stabilization are also discussed. Studies indicate that ACC can be used to construct environmentally-friendly hybrid films, supramolecular hydrogels, and drug vehicles. Finally, the existing challenges and future directions of the controlled synthesis and functionalization of CaCO3 and its expanding applications are highlighted.

GOLPH3/CKAP4 promotes metastasis and tumorigenicity by enhancing the secretion of exosomal WNT3A in non-small-cell lung cancer.

Cancer metastasis is the main cause of mortality associated with non-small-cell lung cancer (NSCLC), accounting for up to 70% of deaths among patients. The mechanisms underlying distal metastasis remain largely unknown. Golgi phosphoprotein 3 (GOLPH3) correlates negatively with overall survival in multiple tumors. In this study, we evaluated the function of GOLPH3 in NSCLC distal metastasis. GOLPH3 was expressed at high levels in samples from patients with NSCLC and was positively associated with clinicopathologic characteristics including clinical stage (P < 0.001), T (P = 0.001), N (P = 0.007), and M (P = 0.001) classification. Functionally, Transwell and wound-healing assays suggested that GOLPH3 overexpression enhances NSCLC cell migration and invasion abilities. Tumor-sphere formation and flow cytometry assays demonstrated that GOLPH3 overexpression enhances a stem cell-like phenotype of NSCLC cells. Metastasis models established by tail vein and intracardiac injection confirmed the pro-metastatic function of GOLPH3 in vivo. A subcutaneous tumor formation model confirmed that GOLPH3 overexpression increased the tumorigenicity of NSCLC cells. Mechanistically, gene set enrichment analysis revealed a positive association of GOLPH3 mRNA expression with WNT-activated gene signatures. Luciferase-reporter and nuclear extract assays showed that GOLPH3 overexpression enhances metastasis and tumorigenicity through activation of the WNT/ß-catenin pathway. Immunoprecipitation-mass spectrometry and gene ontology analysis demonstrated that GOLPH3 interacts with cytoskeleton-associated protein 4 (CKAP4) in exosome-mediated distal metastasis. We found that GOLPH3 decreased the amount of plasma membrane-localized CKAP4 and increased the amount of exosome-localized CKAP4 to promote the formation of CKAP4-containing exosomes. Furthermore, we demonstrated that CKAP4 binds exosomal WNT3A to enhance its secretion. Therefore, the GOLPH3/CKAP4 axis plays a crucial role in promoting exosomal-WNT3A secretion to enhance and maintain the stem-like phenotype and metastasis in NSCLC, thus indicating the therapeutic potential of GOLPH3 in patients with NSCLC metastasis.

Inclusion of organic species in exfoliated montmorillonite nanolayers towards hierarchical functional inorganic-organic nanostructures.

Montmorillonite (Mt) can readily undergo spontaneous delamination or exfoliation into nanolayers by various physical and chemical processes, which allow various strategies to engineer hierarchical functional inorganic-organic nanostructures. This review aims to discuss the recent progress in the liquid-phase exfoliation of Mt into individual nanolayers and the inclusion chemistry of functional organic species, ions, or molecules into the exfoliated Mt nanolayers to produce hierarchical functional inorganic-organic nanostructures. The exfoliation methods include mechanical force, ultrasonication, and intercalation-assisted exfoliation. Techniques for quickly assessing the quality of the exfoliated Mt nanolayers are still needed. Layer-by-layer (LbL) deposition, template, and evaporation-induced inclusions are examined to fabricate hierarchical Mt-organic species nanocomposites with unique functionalities and properties. The nanocomposites can be produced as multilayered porous films, brick-and-mortar coatings, hydrogels with a house-of-cards structure, core-shell materials, and hollow and mesoporous spherical nanocomposites, which exhibit significant potential for adsorption, catalysis, targeted delivery and controlled drug release, highly sensitive sensors, flame retardant coatings, and thermal energy storage and release (i.e. phase change materials). Finally, the challenges and prospects for the future development of hierarchical nanocomposites of exfoliated Mt nanolayers and organic species, particularly in hierarchical supramolecular nanostructured composites, are highlighted.

SRPK1/AKT axis promotes oxaliplatin-induced anti-apoptosis via NF-κB activation in colon cancer.

BACKGROUND: Colorectal cancer is the third most common diagnosis. Oxaliplatin is used as first-line treatment of colon cancer. However, oxaliplatin resistance greatly reduces its therapeutic effect. SRPK1 involves in pre-mRNA splicing and tumorigenesis. How SRPK1 mediates drug resistance in colon cancer is unknown. METHODS: The expression of SRPK1 was analyzed in the TCGA and the CPTAC pan-cancer samples and detected in colon cancer cell lines and tissues by IHC and western blot. The MTT and TUNEL assay were used to verify the anti-apoptosis ability of colon cancer cell. The activation of NF-κB was determined by luciferase assay and qRT-PCR. AKT, IKK, IκB and their phosphorylation level were verified by western blot. RESULTS: We found that SRPK1 expression was the second highest in TCGA and the CPTAC pan-cancer samples. The mRNA and protein levels of SRPK1 were increased in tissues from patients with colon cancer. SRPK1 was associated with clinical stage and TNM classifications in 148 cases of colon cancer patients. High SRPK1 levels correlated with poor prognosis (p < 0.001). SRPK1 overexpression enhanced the anti-apoptosis ability of colon cancer cells, whereas SRPK1 silencing had the opposite effect under oxaliplatin treatment. Mechanistically, SRPK1 enhances IKK kinase and IκB phosphorylation to promote NF-κB nuclear translocation to confer oxaliplatin resistance. CONCLUSIONS: Our findings suggest that SRPK1 participates in colon cancer progression and enhances the anti-apoptosis capacity to induce drug resistance in colon cancer cells via NF-κB pathway activation, and thus might be a potential pharmaceutically target for colon cancer treatment.

Nanoclay-based drug delivery systems and their therapeutic potentials.

Safe, therapeutically effective, and patient-compliant drug delivery systems are needed to design novel tools and strategies to combat the deadliest of diseases such as cancer, SARS, H7N9 avian influenza, and dengue infection. The major challenges in drug delivery are cytotoxicity, poor biodistribution, insufficient functionality, ineffective drug incorporation in delivery devices, and subsequent drug release. Clay minerals are a class of nanolayered silicates that have good biocompatibility, high specific surface area, chemical inertness, colloid, and thixotropy, and are attractive practical and potential nanomaterials in medicine. These properties enable the usage of nanoclays as drug carriers for the delivery of antibiotics, antihypertensive drugs, anti-psychotic, and anticancer drugs. The review examines the latest advances in nanoclay-based drug delivery systems and related applications in gene therapy and tissue engineering. Clay minerals, particularly montmorillonite, kaolinite, and halloysite are used to delay and/or target drug release or even improve drug dissolution due to their surface charge. Chemical modification of clay minerals such as intercalation of ions into the interlayer space of clay minerals or surface modification of clay minerals is a strategy to tune the properties of nanoclays for the loading and release of a drug. The modified nanoclay can take up drugs by encapsulation, immobilization, ion exchange reaction, or electrostatic interactions. Controlled drug release from the drug-clay originates from the incorporation and interactions between the drug and inorganic layers, including electrostatic interactions and hydrogen bonding. Montmorillonite has proven non-toxic through hematological, biochemical, and histopathological analyses in rat. Montmorillonite can also act as a potent detoxifier. Halloysite nanotubes can bind synthetic and biological components such as chitosan, gelatin, and alginate innate nanocarriers for the improved loading and controlled release of drugs, proteins, and DNA. The peculiar properties of clay nanoparticles lead to promising applications in drug delivery, gene delivery, tissue engineering, cancer and stem cell isolation, and bioimaging.

Improved lead removal from aqueous solution using novel porous bentonite - and calcite-biochar composite.

Biochar-mineral (bentonite/calcite) composite (BC-CM) prepared at different temperatures were tested under varied conditions for effective removal of lead (Pb) from aqueous solution. With increasing pyrolysis temperature, increased surface area, pore volume, bentonite decomposition and less or no decomposition of calcite occurred. Bentonite-biochar (BCS) and calcite-biochar (CCS) prepared at 700 °C were found most suitable for efficient removal of Pb (99.9%). Bentonite and calcite acted as catalyst and contributed to changes in yield, pH, texture, functional groups, minerals and carbonization that facilitated efficient Pb removal by BCS 700 and CCS 700. Pb concentration, pH, dose of BCS and CCS, and contact time were further optimized using response surface methodology (RSM) for maximizing removal percentage (R%) of Pb and adsorption capacity (qt). Both BCS 700 and CCS 700 showed similar effects (positive/negative) of factors on R% and qt. Under optimized conditions, 0.21 g of BCS 700 effectively removed 99.2% of 431 mg/L in 3.6 h at solution pH of 4.2, while 0.07 g CCS 700 removed 97.06% of 232 mg/L in 3.5 h at 5.5 pH. Removal of Pb onto both BCS and CCS was by monolayer adsorption with maximum adsorption capacity of 500 mg/g. Rapid Pb removal was observed within 2 h of contact time (CCS 700 > BCS 700) and equilibrium was achieved within 10 h. BCS 700 followed first order and CCS 700 followed second order kinetic model. Electrostatic attraction between Pb ions and mineral groups present in BCS 700 and CCS 700 also played important role in Pb removal. This study clearly demonstrated that composite of biochar with bentonite or calcite under optimized conditions significantly improved Pb removal and adsorption capacity that can be further utilized for larger scale applications.

A Predictive Scoring Model for Short-Term Local Recurrent Nasopharyngeal Carcinoma Based on Magnetic Resonance Imaging.

OBJECTIVE: To predict the early identification of recurrence based on magnetic resonance imaging (MRI) in nasopharyngeal cancer (NPC) patients. METHODS: The clinical and MRI data of 215 patients with local recurrent NPC were retrospectively reviewed. Logistic regression analysis was performed to distinguish the independent risk factors for the short-term (less than 24 months) local recurrence of NPC. The predictive score model was based on the regression coefficients of significant independent variables. RESULTS: Residual disease in the nasopharyngeal cavity (NC), masticator space invasion (MSI), skull base bone erosion (SBBE), and MRI-detected cranial nerve invasion (MDCNI) were all significant independent risk factors for the short-term recurrence of NPC (p < 0.05). The receiver operating characteristic curve showed that the total score had a maximal AUC (area under the curve) value of 0.897, with a cutoff point of 10.50. The sensitivity and specificity were 79.4% and 80.5%, respectively. CONCLUSION: Residual lesions in NC, MSI, SBBE, and MDCNI are independent risk factors in predicting the short-term recurrence of NPC. The authors' findings suggest that patients with a score of more than 10.50 points should be hypervigilant regarding the possibility of short-term recurrence.

Capture and recycling of ammonium by dolomite-aided struvite precipitation and thermolysis.

The capture and reuse of NH4+ is an ideal solution to treat NH4+-containing wastewater. The capture and reuse process needs to be clean and cost-effective. Currently, however, there are many obstacles, particularly in the availability, cost, and recovery of the chemical sources required. Here, we demonstrate a clean and efficient method to capture and recycle NH4+ by a dolomite-aided struvite precipitation process. Dolomite calcined carefully in CO2 atmosphere was used as a Mg source to react with PO43- (KH2PO4) and NH4+ in model wastewater (2000 mg L-1 NH4+). The precipitation was performed at nMg2+:nNH4+:nPO43- = 1:1:1.2 and pH = 8.0 for 2 h; 89.7% of NH4+ was recovered in the form of struvite precipitate. The competition between K+ and NH4+ in the model wastewater led to the formation of K-struvite (MgKPO4·6H2O) and struvite (MgNH4PO4·6H2O). The formation of K-struvite resulted in a decrease in the NH4+ removal rate. When struvite was heated at 110 °C for 4 h, the NH4+ release rate from the thermolysis reached 75.7%. Thermolysis readily occurred as an unstable Ca2+-CO32--NH4+ system formed in the mixture of MgNH4PO4·6H2O and CaCO3. The elements Mg and P that were obtained during the struvite precipitation-thermolysis-reprecipitation process can be repeatedly used. After 6 cycles, under the conditions pH = 9.0, nMg2+:nNH4+:nPO43- = 1:1:1 and reaction time of 2 h, up to 78.3% of NH4+ was removed from the model wastewater.

Recent advances in clay mineral-containing nanocomposite hydrogels.

Clay mineral-containing nanocomposite hydrogels have been proven to have exceptional composition, properties, and applications, and consequently have attracted a significant amount of research effort over the past few years. The objective of this paper is to summarize and evaluate scientific advances in clay mineral-containing nanocomposite hydrogels in terms of their specific preparation, formation mechanisms, properties, and applications, and to identify the prevailing challenges and future directions in the field. The state-of-the-art of existing technologies and insights into the exfoliation of layered clay minerals, in particular montmorillonite and LAPONITE®, are discussed first. The formation and structural characteristics of polymer/clay nanocomposite hydrogels made from in situ free radical polymerization, supramolecular assembly, and freezing-thawing cycles are then examined. Studies indicate that additional hydrogen bonding, electrostatic interactions, coordination bonds, hydrophobic interaction, and even covalent bonds could occur between the clay mineral nanoplatelets and polymer chains, thereby leading to the formation of unique three-dimensional networks. Accordingly, the hydrogels exhibit exceptional optical and mechanical properties, swelling-deswelling behavior, and stimuli-responsiveness, reflecting the remarkable effects of clay minerals. With the pivotal roles of clay minerals in clay mineral-containing nanocomposite hydrogels, the nanocomposite hydrogels possess great potential as superabsorbents, drug vehicles, tissue scaffolds, wound dressing, and biosensors. Future studies should lay emphasis on the formation mechanisms with in-depth insights into interfacial interactions, the tactical functionalization of clay minerals and polymers for desired properties, and expanding of their applications.

MiR-125a-3p regulates glioma apoptosis and invasion by regulating Nrg1.

The current study was designed to examine the functional role and mechanism of miR-125a-3p in glioma development. Quantitative RT-PCR was used to evaluate miR-125a-3p expression in 60 glioma cases of different malignant grades. Then, the clinic pathologic significance of miR-125a-3p expression was determined in combination with the prognosis of the patients. In addition, the effects and mechanisms of miR-125a-3p on the proliferation, apoptosis and invasion of glioma cells were further investigated. The results showed that the expression of miR-125a-3p was decreased significantly in most malignant glioma samples relative to normal brain tissues and glioma tissues of low-malignant degree. Further kaplan-meier survival analysis showed that the lower expression of miR-125a-3p was associated with a poor prognosis of GBM patients. Functional analysis showed that the reintroduction of miR-125a-3p into glioblastoma cell lines induces markedly the apoptosis and suppresses the proliferation and migration of glioblastoma cells in vitro and in vivo. Luciferase assay and Western blot analysis revealed that Nrg1 is a direct target of miR-125a-3p. Furthermore, an increased expression of Nrg1 could reverse the effects of overexpression of miR-125a-3p on the proliferation, apoptosis and migration of glioblastoma cells. These findings suggest that miR-125a-3p performed an important role in glioma development mediated by directly regulating the expression of Nrg1. This study also provides a potential target for diagnosis and treatment of malignant glioma.

Clinical significance of SOX9 in human non-small cell lung cancer progression and overall patient survival.

BACKGROUND: Sex determining region Y (SRY)-related high mobility groupbox 9 (SOX9) is an important transcription factor required for development, which regulates the expression of target genes in the associated pathway. The aim of this study was to describe the expression of SOX9 in human non-small cell lung cancer (NSCLC) and to investigate the association between SOX9 expression and progression of NSCLC. METHODS: SOX9 protein and mRNA expression in normal human pneumonocytes, lung cancer cell lines, and eight pairs of matched lung cancer tissues and their adjacent normal lung tissues were detected by Western blotting and real-time reverse transcription-polymerase chain reaction (RT-PCR). Immunohistochemistry was used to determine SOX9 protein expression in 142 cases of histologically characterized NSCLC. Statistical analyses were applied to test for prognostic and diagnostic associations. RESULTS: SOX9 in lung cancer cell lines was upregulated at both mRNA and protein levels, and SOX9 mRNA and protein were also elevated in NSCLC tissues compared with levels in corresponding adjacent non-cancerous lung tissues. Immunohistochemical analysis demonstrated a high expression of SOX9 in 74/142 (52.1%) paraffin-embedded archival lung cancer biopsies. Statistical analysis indicated that upregulation of SOX9 was significantly correlated with the histological stage of NSCLC (P=0.017) and that patients with a high SOX9 level exhibited a shorter survival time (P<0.001). Multivariate analysis illustrated that SOX9 upregulation might be an independent prognostic indicator for the survival of patients with NSCLC. CONCLUSIONS: This work shows that SOX9 may serve as a novel and prognostic marker for NSCLC, and play a role during the development and progression of the disease.

Catalytic conversion of lignocellulosic biomass to fine chemicals and fuels.

Lignocellulosic biomass is the most abundant and bio-renewable resource with great potential for sustainable production of chemicals and fuels. This critical review provides insights into the state-of the-art accomplishments in the chemocatalytic technologies to generate fuels and value-added chemicals from lignocellulosic biomass, with an emphasis on its major component, cellulose. Catalytic hydrolysis, solvolysis, liquefaction, pyrolysis, gasification, hydrogenolysis and hydrogenation are the major processes presently studied. Regarding catalytic hydrolysis, the acid catalysts cover inorganic or organic acids and various solid acids such as sulfonated carbon, zeolites, heteropolyacids and oxides. Liquefaction and fast pyrolysis of cellulose are primarily conducted over catalysts with proper acidity/basicity. Gasification is typically conducted over supported noble metal catalysts. Reaction conditions, solvents and catalysts are the prime factors that affect the yield and composition of the target products. Most of processes yield a complex mixture, leading to problematic upgrading and separation. An emerging technique is to integrate hydrolysis, liquefaction or pyrolysis with hydrogenation over multifunctional solid catalysts to convert lignocellulosic biomass to value-added fine chemicals and bio-hydrocarbon fuels. And the promising catalysts might be supported transition metal catalysts and zeolite-related materials. There still exist technological barriers that need to be overcome (229 references).

Chemoselective catalytic conversion of glycerol as a biorenewable source to valuable commodity chemicals.

New opportunities for the conversion of glycerol into value-added chemicals have emerged in recent years as a result of glycerol's unique structure, properties, bioavailability, and renewability. Glycerol is currently produced in large amounts during the transesterification of fatty acids into biodiesel and as such represents a useful by-product. This paper provides a comprehensive review and critical analysis on the different reaction pathways for catalytic conversion of glycerol into commodity chemicals, including selective oxidation, selective hydrogenolysis, selective dehydration, pyrolysis and gasification, steam reforming, thermal reduction into syngas, selective transesterification, selective etherification, oligomerization and polymerization, and conversion of glycerol into glycerol carbonate.

[Screening for lymphatic metastasis-associated genes in mouse hepatocarcinoma cell lines Hca-F and Hca-P using gene chip].

BACKGROUND & OBJECTIVE: Metastasis is a complex process involving multiple genetic changes. The high mortality and poor prognosis caused by metastasis in malignant tumor patients and the uncertain mechanisms are always the prominent problems in the field of oncology. In order to screen for lymphatic metastasis-associated genes, the gene expression profiles of mouse hepatocarcinoma cell lines Hca-F (highly metastatic) and Hca-P (low metastatic) were compared by gene chip. METHODS: Total RNA was isolated from Hca-F and Hca-P cells, and synthesized into double-stranded cDNA, then synthesized into biotin-labeled cRNA probes by in vitro transcription. The cRNA probes were separately hybridized with Affymetrix GeneChip MOE430A (containing 22,690 transcripts, including 14,500 known mouse genes and 4,371 ESTs), and the signals were scanned by the GeneArray Scanner. The results were analyzed by bioinformatics. RESULTS: Compared with the gene expression profile of Hca-P cells, 901 (6.2%) genes and 129 (3%) ESTs were up-regulated by at least 2 folds in Hca-F cells; 33 genes, including endoglin (EDG; CD105), Mcam (Muc18; Mel-CAM; CD146), Cdc42ep5 (CEP5; Borg3), Ptprr (protein tyrosine phosphatase, receptor type, R), F2r [coagulation factor II (thrombin) receptor; Par1; ThrR], D7Ertd458e (necl-5), NR1D1, Serpin h1 (HSP47), AXL, Mak, and Areg (AR), were up-regulated 13.93-29.86 folds. According to Gene Ontology and Treeview analysis, these 33 genes were involved in angiogenesis, cell adhesion, signal transduction, cell motility, transcription, chaperone activity, protein kinase activity, receptor binding, and so on. CONCLUSION: Many lymphatic metastasis-associated genes were screened by high-throughput gene chip method; validating their cellular functions will help to identify the key or candidate gene/pathway responsible for lymphatic metastasis, which might be used as diagnostic markers and therapeutic targets for lymphatic metastasis.

Identify lymphatic metastasis-associated genes in mouse hepatocarcinoma cell lines using gene chip.

AIM: In order to obtain lymphogenous metastasis-associated genes, we compared the transcriptional profiles of mouse hepatocarcinoma cell lines Hca-F with highly lymphatic metastasis potential and Hca-P with low lymphatic metastasis potential. METHODS: Total RNA was isolated from Hca-F and Hca-P cells and synthesized into double-stranded cDNA. In vitro transcription double-stranded cDNA was labeled with biotin (i.e. biotin-labeled cRNA, used as the probe). The cRNA probes hybridized with Affymetrix GeneChip(r) MOE430A (containing 22 690 transcripts, including 14 500 known mouse genes and 4 371 ESTs) respectively and the signals were scanned by the GeneArray Scanner. The results were then analyzed by bioinformatics. RESULTS: Out of the 14 500 known genes investigated, 110 (0.8%) were up regulated at least 2(3) fold. Among the total 4 371 ESTs, 17 ESTs (0.4%) (data were not presented) were up regulated at least 2(3) fold. According to the Gene Ontology and TreeView analysis, the 110 genes were further classified into two groups: differential biological process profile and molecular function profile. CONCLUSION: Using high-throughput gene chip method, a large number of genes and their cellular functions about angiogenesis, cell adhesion, signal transduction, cell motility, transport, microtubule-based process, cytoskeleton organization and biogenesis, cell cycle, transcription, chaperone activity, motor activity, protein kinase activity, receptor binding and protein binding might be involved in the process of lymphatic metastasis and deserve to be used as potential candidates for further investigation. Cyclin D1, Fosl1, Hsp47, EGFR and AR, and Cav-1 are selected as the possible candidate genes of the metastatic phenotype, which need to be validated in later experiments. ESTs (data were not presented) might indicate novel genes associated with lymphatic metastasis. Validating the function of these genes is helpful to identify the key or candidate gene/pathway responsible for lymphatic metastasis, which might be used as the diagnostic markers and the therapeutic targets for lymphatic metastasis.