Nongyue He

Highly frequent PIK3CA amplification is associated with poor prognosis in gastric cancer

BackgroundThe phosphoinositide 3-kinase (PI3K)/Akt pathway plays a fundamental role in cell proliferation and survival in human tumorigenesis, including gastric cancer. PIK3CA mutations and amplification are two major causes of overactivation of this pathway in human cancers. However, until this work, there was no sound investigation on the association of PIK3CA mutations and amplification with clinical outcome in gastric cancer, particularly the latter.MethodsUsing direct sequencing and real-time quantitative PCR, we examined PIK3CA mutations and amplification, and their association with clinicopathological characteristics and clinical outcome of gastric cancer patients.ResultsPIK3CA mutations and amplification were found in 8/113 (7.1%) and 88/131 (67%) gastric cancer patients, respectively. PIK3CA amplification was closely associated with increased phosphorylated Akt (p-Akt) level. No relationship was found between PIK3CA mutations and clinicopathological characteristics and clinical outcome in gastric cancer. PIK3CA amplification was significantly positively associated with cancer-related death. Importantly, Kaplan-Meier survival curves revealed that the patients with PIK3CA amplification had significantly shorter survival times than the patients without PIK3CA amplification.ConclusionsOur data showed that PIK3CA mutations were not common, but its amplification was very common in gastric cancer and may be a major mechanism in activating the PI3K/Akt pathway in gastric cancer. Importantly, Kaplan-Meier survival curves revealed that PIK3CA amplification was significantly positively associated with poor survival of gastric cancer patients. Collectively, the PI3K/Akt signaling pathway may be an effective therapeutic target in gastric cancer.

Construction of a Glucose Biosensor Immobilized With Glucose Oxidase in the Film of Polypyrrole Nanotubules

By electrochemical adsorption of glucose oxidase (GO) on a polypyrrole (PPy) nanotubular layer, a glucose biosensor has been fabricated. PPy microtubules can be synthesized by oxidative polymerization of the pyrrole monomer within the pores of a polycarbonate template. The activities of immobilized GO are investigated and the electrochemical properties are characterized by comparison of the GO electrode constructed by electropolymerization and doping.

Self-assembly monolayer of mercaptopropyltrimethoxysilane for electroless deposition of Ag

Mercaptopropyltrimethoxysilane (MPTS) was used to form self-assembly monolayers (SAMs) on glass slides, which was verified by using X-ray photoelectron spectroscopy (XPS) and auger electron spectroscopy (AES). Electroless plating of Ag was performed on the SAMs-modified glass slide. XPS study showed that Ag colloids formed in solution were successfully and hard anchored on SAMs through chemical bonds. Scanning electron microscopy (SEM) analysis illustrated that Ag film on the SAMs-modified glass showed more predominant in durability of temperature than that on conventionally modified glass.

Quantitative assessment of gene methylation and their impact on clinical outcome in gastric cancer.

BACKGROUNDnPromoter methylation is an alternative mechanism of gene silencing in human tumorigenesis. Although a number of methylated genes have been found in gastric cancer, useful methylation markers for early diagnosis and prognostic evaluation of this cancer remain largely unknown.nnnMETHODSnUsing quantitative methylation-specific PCR (Q-MSP), we examined promoter methylation of 6 genes, including CALCA, DAPK1, RARbeta, RASSF1A, TIMP3, and PAX6, and explored their association with clinical outcome in gastric cancer.nnnRESULTSnWe found that most of the genes investigated in the present study had significantly higher methylation level in tumor tissues than normal gastric tissues, including CALCA, RARbeta, RASSF1A, TIMP3, and PAX6. With more focus on specificity compared to sensitivity, all genes were hypermethylated in gastric cancer, ranging from 12.8% to 36.9%. Methylation of TIMP3 and PAX6 was strongly associated with differentiation and lymph node metastasis, respectively. Importantly, most of gene methylation, except for DAPK1, was closely associated with poor survival in gastric cancer.nnnCONCLUSIONnWe found that a panel of genes was specifically methylated in gastric cancer, and demonstrated the effect of promoter methylation of some genes on clinical outcome in gastric cancer, indicating these methylated genes may be useful biomarkers for prognostic evaluation in this cancer.

Investigating the state of Fe and La in MCM-41 mesoporous molecular sieve materials

Abstract Fe-containing, La-containing, siliceous and aluminum silicate MCM-41 mesoporous materials were synthesized using water glass as silica source. The states of Fe(III) and La(III) species were investigated by thermogravimetric analysis (TGA), powder X-ray diffraction (XRD), framework infrared (FTIR), electron spin resonance (ESR) and Mossbauer spectroscopic techniques. It was shown that Fe(III) or La(III) species was incorporated into framework. Because the Fe–O or La–O bond is longer than Si–O bond, Fe(III) or La(III) species was limited to insert into framework and transformed from tetrahedrally coordinated state to octahedrally coordinated state upon calcination to remove template.

Frequent Gene Amplification Predicts Poor Prognosis in Gastric Cancer

Gastric cancer is one of the most common malignancies worldwide. However, genetic alterations leading to this disease are largely unknown. Gene amplification is one of the most frequent genetic alterations, which is believed to play a major role in the development and progression of gastric cancer. In the present study, we identified three frequently amplified genes from 30 candidate genes using real-time quantitative PCR method, including ERBB4, C-MET and CD44, and further explored their association with clinicopathological characteristics and poor survival in a cohort of gastric cancers. Our data showed amplification of these genes was significantly associated with certain clinicopathological characteristics, particularly tumor differentiation and cancer-related death. More importantly, amplification of these genes was significantly related to worse survival, suggesting that these amplified genes may be significant predictors of poor prognosis and potential therapeutic targets in gastric cancer. Targeting these genes may thus provide new possibilities in the treatment of gastric cancer.

An electrochemical approach for detection of specific DNA-binding protein by gold nanoparticle-catalyzed silver enhancement

Interaction between transcription factor and sequence-specific DNA plays an important role in regulation of gene transcription in biological systems. As electrochemical intercalators, gold (Au) nanoparticles show high catalysis activity and compatibility for detection of biological molecules. In this article, we report an electrochemical approach for sequence-specific DNA-binding transcription factor detection by Au nanoparticle-catalyzed silver (Ag) enhancement at interface between electrodes and electrolyte solutions. Here unimolecular hairpin oligonucleotides were self-assembled onto Au electrode surface and their elongation on Au electrode surface was carried out to form double-stranded oligonucleotides with transcription factor NF-kappaB (nuclear factor-kappa B) binding sites. Au nanoparticle-catalyzed Ag deposition was detected by anodic stripping voltammetry (ASV) for NF-kappaB binding. It was found that this method for the detection of sequence-specific DNA-binding protein showed pronounced specificity and that the detection limit was as low as 0.1 pM. The findings indicated that our method can have applications in transcription regulation, operator site recognition, and functional gene inspection.

Metallothionein 1G functions as a tumor suppressor in thyroid cancer through modulating the PI3K/Akt signaling pathway

BackgroundMT1G inactivation mediated by promoter methylation has been reported in thyroid cancer. However, the role of MT1G in thyroid carcinogenesis remains unclear. The aim of this study is to examine the biological functions and related molecular mechanisms of MT1G in thyroid cancer.MethodsMethylation-specific PCR (MSP) was performed to analyze promoter methylation of MT1G and its relationship with clinicopathological characteristics of papillary thyroid cancer (PTC) patients. Conventional and real-time quantitative RT-PCR assays were used to evaluate mRNA expression. The functions of ectopic MT1G expression were determined by cell proliferation and colony formation, cell cycle and apoptosis, as well as cell migration and invasion assays.ResultsMT1G expression was frequently silenced or down-regulated in thyroid cancer cell lines, and was also significantly decreased in primary thyroid cancer tissues compared with non-malignant thyroid tissues. Promoter methylation, along with histone modification, contributes to MT1G inactivation in thyroid tumorigenesis. Moreover, our data showed that MT1G hypermethylation was significantly positively associated with lymph node metastasis in PTC patients. Importantly, restoring MT1G expression in thyroid cancer cells dramatically suppressed cell growth and invasiveness, and induced cell cycle arrest and apoptosis through inhibiting phosphorylation of Akt and Rb.ConclusionsWe have for the first time revealed that MT1G appears to be functional tumor suppressor involved in thyroid carcinogenesis mainly through modulating the phosphatidylinositol-3-kinase (PI3K)/Akt pathway and partially through regulating the activity of Rb/E2F pathway in this study.

A microarray to analyze methylation patterns of p16Ink4a gene 5′-CpG islands

OBJECTIVESnAberrant DNA methylation of the CpG site is among the earliest and most frequent alterations in cancer. Several studies suggest that aberrant methylation on the CpG sites of the tumor suppressor gene is closely associated with carcinogenesis. However, large-scale analysis of candidate genes has so far been hampered by the lack of high throughput approach for analyzing methylation patterns. The aim of this study was to develop a new method to analyze methylation patterns of p16(Ink4a) gene.nnnDESIGN AND METHODSnWe selected a 336 bp segment of the 5 untranslated region and the first exon of the p16(Ink4a) gene, as the target sequence, which include the most densely packed CpG fragment of the islands containing 32 CpG sites. A set of oligonucleotide probes was designed to assemble a DNA microarray to discriminate the methylation patterns of several adjacent CpG sites.nnnRESULTSnMethylation patterns of human p16(Ink4a) gene were mapped and the results were validated by bisulphite DNA sequencing. A good reproducibility was observed in several parallel experiments.nnnCONCLUSIONSnThe methylation oligonucleotide microarray can be applied as a useful and powerful tool to map methylation patterns in multiple CpG island sites.

Synthesis and characterization of SiO2/(PMMA/Fe3O4) magnetic nanocomposites.

Magnetic silica nanocomposites (magnetic nanoparticles core coated by silica shell) have the wide promising applications in the biomedical field and usually been prepared based on the famous Stöber process. However, the flocculation of Fe3O4 nanoparticles easily occurs during the silica coating, which limits the amount of magnetic silica particles produced in the Stöber process. In this paper, PMMA/Fe3O4 nanoparticles were used in the Stöber process instead of the nude Fe3O4 nanoparticles. And coating Fe3O4 with PMMA polymer beforehand can prevent magnetic nanoparticles from the aggregation that usually comes from the increasing of ionic strength during the hydrolyzation of tetraethoxysilane (TEOS) by the steric hindrance. The results show that the critical concentration of magnetic nanoparticles can increase from 12 mg/L for nude Fe3O4 nanoparticles to 3 g/L for PMMA/Fe3O4 nanoparticles during the Stöber process. And before the deposition of silica shell, the surface of PMMA/FeO4 nanoparticles had to be further modified by hydrolyzing them in CH3OH/NH3 x H2O mixture solution, which provides the carboxyl groups on their surface to react further with the silanol groups of silicic acid.