Mutations in AXIN2 cause colorectal cancer with defective mismatch repair by activating beta-catenin/TCF signalling.
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Axin Protein↗
Biomedical subjects
C Qian
Publications and source records attributed to C Qian.
Transduction efficacy, antitumoral effect, and toxicity of adenovirus-mediated herpes simplex virus thymidine kinase/ ganciclovir therapy of hepatocellular carcinoma: the woodchuck animal model.
Gene therapy for hepatocellular carcinoma (HCC) has shown some promise, but its evaluation requires relevant experimental models. With this aim, we present an evaluation of the interest of using the woodchuck model of HCC to assess in vivo gene transfer efficiency. We tested the transduction efficacy of the adenoviral vectors directing lacZ gene product expression under the control of the cytomegalovirus and alpha-fetoprotein (AFP) regulatory sequences. We have also investigated whether an adenoviral cytomegalovirus-thymidine kinase (Tk) vector might induce an antitumoral effect in this model. Our results demonstrate that with direct intratumoral and intrahepatic arterial injections, transduction of a significant proportion of tumor cells occurred even in large HCC nodules. Furthermore, due to intra-arterial anastomoses, direct intratumoral injection led to transduction of some noninjected HCC nodules. Moreover, direct intratumoral injection of a herpes simplex virus-1 Tk-encoding vector induced, on ganciclovir administration, a significant antitumoral effect in the two animals evaluated. However, in one animal, massive hepatic failure occurred due to Tk expression in nontumor cells. These results emphasize the need to target the expression of the Tk gene to tumor cells using a hepatoma-specific promoter such as AFP promoter. However, we showed that, in vivo, lacZ expression as driven by the AFP promoter was extremely low, thus emphasizing some potential pitfalls when using this approach. Altogether, our data stress the need to test gene therapy-based strategies in such in vivo animal models of HCC and evaluate gene transduction, expression, and biological activity, as well as its potential toxicity.
Adenoviridae↗
In vivo gene transfer of CD40 ligand into colon cancer cells induces local production of cytokines and chemokines, tumor eradication and protective antitumor immunity.
The interaction between CD40 ligand (CD40L, CD154) and its receptor CD40 on antigen-presenting cells, is essential for the initiation of cell-mediated and humoral immune responses. In this study, we investigated the antitumor effect of in vivo gene transfer of CD40L to tumor cells using an adenoviral vector (AdCMVmCD40L) in a murine CT-26 colon cancer model. We found that injection of AdCMVmCD40L caused tumor regression in a dose-dependent manner. A complete regression of tumor was observed in 81% of mice treated with 10(9) p.f.u. of AdCMVmCD40L. The antitumor effect induced by CD40L was mediated by CD8+ T cells and was associated with the generation of tumor-specific cytolytic T lymphocytes (CTL). Animals that eradicated the tumor were protected against tumor cell rechallenge, and both CD4+ and CD8+ T cells were involved in specific protective immunity. Treatment with AdCMVmCD40L in one tumor nodule also caused complete regression of established tumors at distant sites. The antitumor effect elicited by AdCMVmCD40L was associated with the intratumoral production of IL-12 and IFN-gamma and with an increased intratumoral expression of chemokines such as MIP- 1alpha, MIP-1beta, MIP-2, RANTES, and eotaxin. These data demonstrate that intratumoral injection of AdCMVmCD40L induces a powerful cascade of chemokines and cytokines in the tumor mass and stimulates an efficient antitumor immunity leading to regression of established colon cancer and protection against tumor cell rechallenge.
Adenoviridae↗
A blood-tumor barrier limits gene transfer to experimental liver cancer: the effect of vasoactive compounds.
We have evaluated gene transfer efficiency to tumor nodules in diethylnitrosoamine (DENA)-induced hepatocellular carcinoma (HCC) in rats using adenoviral vectors administered by three different routes: intraportal, intra-arterial and intratumoral injection. Our results showed that intraportal infusion could not transduce tumor nodules greater than 1 mm in diameter while the intra-arterial route allowed transduction of nodules up to 2-5 mm in diameter. Tumors greater than this size were resistant to transduction by intravascular route, but could be transduced by direct intratumoral injection, indicating that the obstacle preventing gene transfer to tumor cells was mainly at the level of tumor vasculature and not at the level of neoplastic cells. We have studied the extracellular matrix in tumoral lesions to assess whether nodules with different size and histological pattern have different profiles in relation to transduction efficacy. Immunohistochemical detection showed a high expression of fibronectin (FN), laminin (LN) and alpha-smooth muscle actin (alpha-SMA) in those large HCC, which were resistant to adenoviral infection. Intra-arterial infusion of vasoactive compounds (histamine, angiotensin II or nitric oxide donor nitroglycerin) before vector administration enhanced gene transfer to tumor nodules that were poorly transduced without pre-treatment. Nitroglycerin was active to enhance transduction of large tumors with trabecular or pseudoglandular histological pattern, which were impermeable to adenoviral vectors even after histamine or angiotensin treatments. Our data indicate the presence of a physical barrier between blood and neoplastic cells, which prevents transduction of the tumor by vectors given by the intravascular route. The thickness and impermeability of the barrier increases as the tumor nodule grows. Vasoactive compounds may be of value in gene therapy of liver cancer by increasing transduction efficiency by intravascularly administered vectors.
Actins↗
Molecular regulation of constitutive expression of interleukin-8 in human pancreatic adenocarcinoma.
Recent studies have shown that interleukin-8 (IL-8) plays an important role in the growth and metastasis of human pancreatic cancer. In the present study, we determined the molecular regulation of constitutive IL-8 expression in human pancreatic cancer cells. Various human pancreatic cancer cell lines were incubated in vitro. Sixty-seven percent of the cell lines constitutively secreted high levels of IL-8, as determined using enzyme-linked immunosorbent assay. Consistently, these cells constitutively expressed high levels of IL-8 mRNA, as determined using Northern blot analysis. To determine the mechanisms of the high steady-state levels of IL-8 mRNA, the IL-8 half-life and transcription rate were measured. There was no significant difference in IL-8 half-life between cells expressing high and low levels of IL-8. However, higher transcription rates and increased IL-8 promoter activity were observed in the cells constitutively expressing high levels of IL-8. Detailed IL-8 promoter analysis using deletion mutation revealed that the region from -85 to -133 bp was essential for the constitutive IL-8 promoter activity. Also, point-mutation analysis indicated that mutation of NF-kappaB, AP-1, or NF-IL-6 binding sites significantly reduced or eliminated the constitutive IL-8 promoter activity. Consistent with the constitutive IL-8 transcription activity, high levels of constitutive NF-kappaB and AP-1 activity were detected in the cells overexpressing IL-8, as determined using electrophoretic mobility shift assay. In addition, transfection of a dominant-negative I-kappaBalpha expression vector (I-kappaBalphaM) inhibited constitutive NF-kappaB activity and IL-8 expression in pancreatic cancer cells. Collectively, our data demonstrated that constitutive NF-kappaB and AP-1 activation contributes to the overexpression of IL-8, which in turn plays an important role in tumor angiogenesis and contributes to the aggressive biology of human pancreatic cancer.
Adenocarcinoma↗
An optical model of the human retinal nerve fiber layer: implications of directional reflectance for variability of clinical measurements.
PURPOSE: The reflectance of the retinal nerve fiber layer is highly directional--that is, it depends strongly on the angles of illumination and viewing. This study explored and illustrated the implications of this directional reflectance for nerve fiber layer measurements in the human eye. METHODS: The retina was modeled as a sphere centered on the optic axis of a schematic eye. Nerve fiber ribbons were projected onto the retina and cylindrical light scattering was calculated along each ribbon. The reflectance along the ribbon was then determined for the illuminating and viewing apertures of two hypothetical optical instruments, a fundus camera and a confocal scanning laser ophthalmoscope. Results were displayed as reflectance maps. RESULTS: Uniformly illuminated nerve fiber ribbons exhibited a nonuniform reflectance pattern that was very sensitive to the location in the pupil of the instrument apertures. Ribbon reflectance at the superior and inferior disc margins varied with ribbon orientation, being higher with temporal tilt and lower with nasal tilt. Ribbons nasal to the disk could be quite dim. CONCLUSIONS: In quantitative nerve fiber layer assessment technologies, the observed reflectance depends on the configuration of the illuminating and viewing apertures of the measuring instrument and on the retinal position and orientation of each nerve fiber bundle. In clinical practice, this dependence may cause significant measurement variability that can be reduced by specific measurement maneuvers.
Fundus Oculi↗
A rational approach towards successful crystallization and crystal treatment of human cytomegalovirus protease and its inhibitor complex.
The crystallization and subsequent crystal treatment of both free human cytomegalovirus (hCMV) protease and its inhibitor complexes are reported. For free-enzyme crystals, diffraction was greatly improved by optimizing the crystallization conditions, raising the precipitant concentration in the reservoir and soaking the crystals in artificial mother liquors. Each of the six components in the final crystallization formula (16% PEG 4000, 0.1 M MES pH 6.0, 0.4 M LiCl, 10% glycerol, 5% t-butanol and 5 mM Na(2)S(2)O(3)) plays a distinctive role and is indispensable. A synergistic effect of Na(2)SO(4) and t--butanol on diffraction was observed and studied. A 2.0 A multiwavelength anomalous diffraction (MAD) data set was collected using a synchrotron-radiation source, leading to the elucidation of the three-dimensional structure of the enzyme. For the inhibitor-complex crystals, initial attempts with co-crystallization and soaking experiments at pH 6.0 did not produce conclusive results. Subsequently, experiments were designed to co-crystallize the complex at pH 8.0, the optimal pH for the enzyme and the inhibitor activity. Using 20-50 mM spermine in the crystallization buffer, crystals of two peptidomimetic inhibitor complexes were obtained at pH 7.5 and 8.0. Spermine was required for the inhibitor complexes to be crystallized at pH 8.0, possibly neutralizing net negative charges of hCMV protease owing to its acidic pI of 5.5. A 2.7 A data set was collected from one of the inhibitor complexes and the structure was determined using the molecular-replacement method.
Crystallization↗
Method to decrease the titers of contaminating helper adenovirus during the production of recombinant adeno-associated virus.
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Dependovirus↗
Differential expression and allelotyping of the p73 gene in neuroblastoma.
p73 has recently been identified as a candidate imprinted tumor suppressor gene in neuroblastoma. To determine the possible involvement of this gene in the pathogenesis of neuroblastoma, we analyzed allelic expression, screened for mutations and determined MYCN copy numbers in 31 primary neuroblastoma tumor samples. Interestingly, the gene was biallelically expressed in 50% (4/8) of informative neuroblastomas, which suggests that activation of the normally silenced allele of this gene plays an important role in the tumorigenesis of neuroblastoma. However, no tumor-specific mutations were identified although 15 polymorphisms were detected in this gene. We also detected a very strong association between a C91T polymorphism and MYCN copy number in this tumor. The T allele was detected in 8/17 (47%) neuroblastomas without MYCN amplifications but not detected in cases with MYCN amplifications (0/14). The biological significance of this association, however, is unknown. Overall the data suggest that p73 may play an important role in the pathogenesis of neuroblastoma but that the true tumor suppressor gene localized to this area still remains to be identified.
Alleles↗
[Analysis of the interaction of the polymorphisms of presenilin-1 gene and ApoE gene in Alzheimerns disease].
OBJECTIVE: To explore the interaction of the polymorphism of 3' -terminal intron of the eighth exon of presenilin-1(PS-1) gene with the polymorphism of apolipoprotein E(ApoE) gene in the pathogenesis of sporadic Alzheimerns disease(SAD). METHODS: The polymorphisms of 3'-terminal intron of the eighth exon of presenilin-1 gene and ApoE gene were detected by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique in 75 patients with SAD and 73 normal controls. RESULTS: The polymorphisms of PS-1 gene and ApoE gene were associated obviously with SAD. The onset of SAD was negatively associated with allele 2 of PS-1 gene and E(2) allele of ApoE gene,and it was positively associated with allele 1 of PS-1 gene. The onset of SAD was not associated with E(3) allele and E(4) allele of ApoE gene. CONCLUSION: These results indicated that allele 2 of PS-1 gene and E(2) allele of ApoE gene might protect people from SAD, that allele 1 of PS-1 gene might increase the risk of the onset of SAD, and that the onset of SAD was not associated with the interaction of the polymorphisms of PS-1 gene and ApoE gene.
Aged↗
[Gene therapy for liver diseases].
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Adenoviridae↗
[Cloning and expression of a resistant Culex gene in Escherichia coli].
Recombinant plasmid pRLB1 was constructed from detoxifying gene B1 of pesticide resistant Culex pipiens quinquefasciatus and from plasmid pRL439 contained the strong promoter PpsbA. The positive clone was identified by digestion and Southern analysis. Expression of recombinant plasmid containing esterase gene was detected. An engineered bacterium possessing high enzyme activity was obtained and immobilized. It can effectively degrade the specific substrate alpha-naphthyl acetate (alpha-NA) and beta-naphthyl of esterase enzyme. Assays showed that pesticide acetofenate (7504 an organic choride pesticide) was degraded by the immobilized cells within one hour.
Animals↗
Different doses of adenoviral vector expressing IL-12 enhance or depress the immune response to a coadministered antigen: the role of nitric oxide.
Joint immunization with two recombinant adenoviruses, one expressing hepatitis C virus (HCV) core and E1 proteins and another expressing IL-12 (RAdIL-12), strongly potentiates cellular immune response against HCV Ags in BALB/c mice when RAdIL-12 was used at doses of 1 x 105-1 x 107 plaque-forming units. However, cellular immunity against HCV Ags was abolished when higher doses (1 x 108 plaque-forming units) of RAdIL-12 were used. This immunosuppressive effect was associated with marked elevation of IFN-gamma and nitric oxide in the serum and increased cell apoptosis in the spleen. Administration of N-nitro-L -arginine methyl ester (L-NAME), an inhibitor of nitric oxide synthase, to mice that received high doses of RAdIL-12 was lethal, whereas no apparent systemic toxicity by L -NAME was observed in those immunized with lower doses of the adenovirus. Interestingly, in mice immunized with recombinant adenovirus expressing core and E1 proteins of HCV in combination with RAdIL-12 at low doses (1 x 107 plaque-forming units), L -NAME inhibited T cell proliferation and CTL activity in response to HCV Ags and also production of Abs against adenoviral proteins. In conclusion, gene transfer of IL-12 can increase or abolish cell immunity against an Ag depending of the dose of the vector expressing the cytokine. IL-12 stimulates the synthesis of NO which is needed for the immunostimulating effects of IL-12, but apoptosis of T cells and immunosuppression ensues when IFN-gamma and NO are generated at very high concentrations.
Adenoviridae↗
Cloning of the human homolog of conductin (AXIN2), a gene mapping to chromosome 17q23-q24.
Conductin or Axil, an Axin homolog, plays an important role in the regulation of beta-catenin stability in the Wnt signaling pathway. To facilitate the molecular analysis of the human gene, we isolated the human homolog, AXIN2. The cDNA contains a 2529-bp open reading frame and encodes a putative protein of 843 amino acids. Compared with rat and mouse homologs, AXIN2 shows an overall 89% amino acid identity. Several functional domains in this protein are highly conserved including the GRS (95.9%), GSK-3beta (96.3%), Dsh (98%), and beta-catenin (89.9%) domains. Radiation hybrid mapping localized the AXIN2 gene to human chromosome 17q23-q24, a region that shows frequent loss of heterozygosity in breast cancer, neuroblastoma, and other tumors. Human AXIN2 is thus a very strong candidate involved in multiple tumor types.
Adaptor Proteins, Signal Transducing↗
Intratumoral injection of bone-marrow derived dendritic cells engineered to produce interleukin-12 induces complete regression of established murine transplantable colon adenocarcinomas.
Stimulation of the antitumor immune response by dendritic cells (DC) is critically dependent on their tightly regulated ability to produce interleukin-12 (IL-12). To enhance this effect artificially, bone marrow (BM)-derived DC were genetically engineered to produce high levels of functional IL-12 by ex vivo infection with a recombinant defective adenovirus (AdCMVIL-12). DC-expressing IL-12 injected into the malignant tissue eradicated 50-100% well established malignant nodules derived from the injection of two murine colon adenocarcinoma cell lines. Successful therapy was dependent on IL-12 transfection and was mediated only by syngeneic, but not allogeneic BM-derived DC, indicating that compatible antigen-presenting molecules were required. The antitumor effect was inhibited by in vivo depletion of CD8+ T cells and completely abrogated by simultaneous depletion with anti-CD4 and anti-CD8 mAbs. Mice which had undergone tumor regression remained immune to a rechallenge with tumor cells, showing the achievement of long-lasting systemic immunity that also was able to reject simultaneously induced concomitant untreated tumors. Tumor regression was associated with a detectable CTL response directed against tumor-specific antigens probably captured by DC artificially released inside tumor nodules. Our results open the possibility of similarly treating the corresponding human malignancies.
Adenocarcinoma↗
Gene therapy of viral hepatitis and hepatocellular carcinoma.
Gene therapy represents an attractive approach to treat a great variety of diseases, both inherited and acquired, and it is moving slowly from a proof-of-principle phase to a wide application in most medical fields. Liver cancer and viral hepatitis are natural targets for this new therapeutic alternative due to the lack of success of conventional antitumoral and antiviral treatments and the ominous prognosis related with liver tumours. Gene therapy for viral hepatitis is aimed to boost the patient immune response against viral antigens or to make cells resistant to infection by blocking the viral life cycle. Gene transfer techniques applied to the treatment of hepatocellular carcinoma include drug sensitization by suicide genes, genetic immunotherapy, normal tissue protection by transfer of the multidrug resistance gene, replacement of tumour suppressor genes, inhibition of oncogenes and modifications of the biology of the tumour (antiangiogenesis). However, major advances in our understanding of the regulation of gene expression, design of the expression cassettes and development of more efficient gene transfer vectors are mandatory before gene therapy can become a widely used therapeutic modality.
Carcinoma, Hepatocellular↗
[A study on mutation of exon 5 of presenilin-1 in Alzheimer's disease].
OBJECTIVE: To explore the role of the mutation of presenilin-1 in pathogenesis of sporadic Alzheimer's disease. METHODS: The exon 5 of presenilin-1 was analysed by using polymerase chain reaction- single strand conformation polymorphism (PCR-SSCP) and ABI 310 genetic analyzer technique in 68 patients with Alzheimer's disease and 65 normal controls. RESULTS: A mobility shift of SSCP in exon 5 of presenilin-1 was detected in 4 patients with Alzheimer's disease;two missense mutations were found. One mutation was at Leu 130 Met and the other at Val157 Leu. Another mobility shift of SSCP in exon 5 of presenilin-1 was detected in 11 patients with Alzheimer's disease. One missense mutation was at Leu 130 Met and one same sense mutation C462-->T. But no mobility shift of SSCP was found in patients with vascular dementia and in normal controls. CONCLUSION: The authors found that mutations in exon 5 of presenilin-1 also existed in patients with sporadic Alzheimer's disease, which might be one of the points of mutations in sporadic Alzheimer's disease in Chinese.
Aged↗
[The effects of reactive oxidants on Bacillus thuringiensis parasporal crystals].
The effects of hydrogen peroxides(H2O2) and hydroxyl radicals(.OH) on the purified parasporal crystals from Bacillus thuringiensis var. Kurstaki HD-1 were studied by SDS-PAGE and bioassay. The results showed that to some extent H2O2 and .OH produced a direct and deleterious effect. Hydroxyl radicals from the system Fe(II)-EDTA-H2O2 indicated more serious damage than hydrogen peroxides. There was a 44.5% loss in activity against 12-h-old larvae of Heliothis armigera when parasporal crystals were treated with 0.3% H2O2 only. However, 85.2% activity lost when parasporal crystals were treated with 0.3% H2O2 in the presence of iron(II) and EDTA.
Animals↗