ABSTRACT
The receptor for advanced glycation end products (RAGE) has previously been suggested to stimulate the growth, survival, and metastatic spread of colorectal cancers (CRC). The genetic variant Gly82Ser of RAGE influences its function and is associated with an increased risk of gastric cancer and multiple sclerosis. To investigate the association between the Gly82Ser polymorphisms of RAGE and the risk of CRC, 90 CRC patients and 78 control subjects with benign polyps were genotyped and the results were analyzed using the SPSS statistical software.In comparing with the control group, the CRC group has a higher ratio in the Gly82Ser polymorphism. The odds ratio (OR) for heterozygous GS is 2.037 (95% CI 1.207-3.438); the OR for carriers with the S allele (SS) is 3.32 (95% CI 0.94-11.65). Further stratification analysis of the correlation of the Gly82Ser polymorphism with tumor stages and differentiation indicated that CRC patients with TNM (III + IV) and/or patients with poorly differentiated colorectal cancer have an elevated Gly82Ser polymorphism. The OR for TNM (III + IV) is 3.575, 95% CI 1.495-8.550, and the OR for poorly differentiated is 3.580, 95% CI 1.390-9.217. In conclusion, the RAGE gene Gly82Ser polymorphism may confer not only an increased risk of CRC but also an increased invasion of CRC in the Chinese population.
Subject(s)
Colorectal Neoplasms/genetics , Polymorphism, Genetic , Receptors, Immunologic/genetics , Adult , Aged , Aged, 80 and over , Base Sequence , Colorectal Neoplasms/etiology , Colorectal Neoplasms/pathology , Female , Genotype , Humans , Male , Middle Aged , Molecular Sequence Data , Neoplasm Staging , Receptor for Advanced Glycation End Products , RiskABSTRACT
UNLABELLED: The EphA5 receptor has recently been known to play an important role in the initiation of the early phase of synaptogenesis, during which irreparable harm would be done to the developing brain in the absence of sufficient thyroid hormone (TH). In the present article, we aimed to analyze the characteristics of EphA5 receptor expression in the brain of congenital hypothyroid rats. The results showed that the levels of the EphA5 receptor were downregulated by TH deficiency in the developing rat brain with remarkable spatial and temporal characteristics. In the hypothyroid rats, the mRNA and protein levels of EphA5 receptor decreased significantly in the hippocampus (27.92-53.26%), cerebral cortex (12.52-47.16%), and cerebellum (8.72-31.69%) compared with those in the normal rats from postnatal day 0 (P0) to P21 (p < 0.01). The expression of EphA5 receptor was highest and declined most as much as 53% in the hippocampus with TH deficiency. At P7, the EphA5 receptor decreased most prominently during all the observed time point. CONCLUSION: The EphA5 receptor plays actively in the brain development in congenital hypothyroid rats. Our study highlights the high expression of EphA5 receptor protein in hippocampus and dramatic changes at P7 in condition of TH deficiency, which may provide important basis for further investigations in manipulating congenital hypothyroidism.
Subject(s)
Brain/metabolism , Congenital Hypothyroidism/metabolism , Hypothyroidism/chemically induced , Receptor, EphA5/metabolism , Thyroid Hormones/metabolism , Animals , Antithyroid Agents , Brain/growth & development , Congenital Hypothyroidism/genetics , Disease Models, Animal , Female , Fluorescent Antibody Technique , Gene Expression , Hypothyroidism/metabolism , Male , Methimazole , RNA, Messenger/metabolism , Rats , Rats, Sprague-Dawley , Real-Time Polymerase Chain Reaction , Receptor, EphA5/geneticsSubject(s)
Doxorubicin/analogs & derivatives , Precursor Cell Lymphoblastic Leukemia-Lymphoma/pathology , Valproic Acid/pharmacology , Apoptosis/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Doxorubicin/administration & dosage , Doxorubicin/pharmacology , Humans , Precursor Cell Lymphoblastic Leukemia-Lymphoma/drug therapy , Valproic Acid/administration & dosageABSTRACT
OBJECTIVE: To study the effect of pGCsi-H1-APRIL on the growth of human colorectal cancer cells in transplated tumor in nude mice and to improve the effect of APRIL on proliferation and apoptosis of colorectal cancer (CRC). METHODS: Human CRC model was established in nude mice, and the nude mice were treated with APRIL siRNA twice per week for 2 weeks. APRIL mRNA expression was surveyed by PCR and APRIL protein expression was detected by immunohistochemistry. The expression of PCNA protein was detected by ELISA. The expression of bcl-2 and bcl-xl was assessed by immunohistochemical staining, and TUNEL staining was used to detect apoptosis. RESULTS: The expression of APRIL mRNA in the APRIL siRNA group was (0.13 ± 0.05) × 10(-3), significantly lower than that in the vector group (0.95 ± 0.04) × 10(-3) and the PBS group (0.96 ± 0.05) × 10(-3). The expression of APRIL protein in the APRIL siRNA group was (87.5 ± 5.0)% lower than that in the vector and PBS groups (P < 0.05). APRIL siRNA significantly suppressed the growth of SW480 tumor: the IR (inhibitory rate) of APRIL siRNA group was (60.7 ± 1.5)% (P < 0.05). The expression of PCNA in APRIL siRNA group was (176.8 ± 18.1) ng/ml, was (56.5 ± 2.0)% lower than that of PBS group (328.4 ± 22.8) ng/ml. Furthermore, the expressions of anti-apoptosis proteins bcl-2 and bcl-xl of APRIL siRNA group were (82.6 ± 4.5)% and (79.2 ± 3.5)% lower than those of the PBS group. The apoptotic rate of the APRIL siRNA group was 40.1% ± 2.5%, significantly higher than that in the vector group (2.5 ± 0.1)% and PBS group (2.5 ± 0.2)% (P < 0.05). CONCLUSION: APRIL siRNA may significantly suppress the growth and promote apoptosis in transplanted tumor of human colorectal cancer in nude mice. APRIL may become a candidate gene of gene therapy of human colorectal cancer.
Subject(s)
Apoptosis , Cell Proliferation , Colorectal Neoplasms/pathology , RNA, Small Interfering/genetics , Tumor Necrosis Factor Ligand Superfamily Member 13/biosynthesis , Animals , Cell Line, Tumor , Colorectal Neoplasms/genetics , Colorectal Neoplasms/metabolism , Female , Humans , Ligands , Mice , Mice, Inbred BALB C , Mice, Nude , Neoplasm Transplantation , Proliferating Cell Nuclear Antigen/metabolism , Proto-Oncogene Proteins c-bcl-2/metabolism , RNA, Messenger/metabolism , Random Allocation , Tumor Necrosis Factor Ligand Superfamily Member 13/genetics , bcl-X Protein/metabolismABSTRACT
The receptor for advanced glycation end products (RAGE) interacts with several ligands and is involved in various human diseases. Several splicing forms of the RAGE gene have been characterized, and two general mechanisms are usually responsible for the generation of soluble receptors. However, variants distribution and respective roles in different tumors are not clear. We analyzed RAGE and hRAGEsec mRNA expression in esophageal and lung cancer by RT-polymerase chain reaction. The Agilent clipper 1000 Bioanalyzer using lab-on-a-chip technology was applied to size and quantify the polymerase chain reaction products. Western blotting was performed to measure total soluble RAGE protein levels. The results showed that RAGE and its splice variants increased in esophageal cancers and decreased in lung cancers. We conclude that RAGE presents as a major isoform; soluble RAGE may also play certain roles in esophageal cancer and lung cancer.
Subject(s)
Esophageal Neoplasms/metabolism , Gene Expression Regulation, Neoplastic , Lung Neoplasms/metabolism , Receptors, Immunologic/metabolism , Aged , Alternative Splicing , Blotting, Western , Computational Biology , Down-Regulation , Electrophoresis , Esophageal Neoplasms/genetics , Esophageal Neoplasms/pathology , Female , Gene Expression Profiling , Humans , Ligands , Lung Neoplasms/genetics , Lung Neoplasms/pathology , Male , Microfluidic Analytical Techniques , Middle Aged , Protein Isoforms/genetics , Protein Isoforms/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , Receptor for Advanced Glycation End Products , Receptors, Immunologic/genetics , Reverse Transcriptase Polymerase Chain Reaction , Solubility , Tissue Distribution , Up-RegulationABSTRACT
OBJECTIVE: To establish a real-time fluorescence quantitative polymerase chain reaction (RFQ-PCR) method for quantifying a proliferation-inducing ligand (APRIL) mRNA in sputum samples from patients with non-small cell lung cancer (NSCLC), and to evaluate its role in the diagnosis of NSCLC. METHODS: Seventy-one cases of NSCLC and 62 cases of benign pulmonary disease were enrolled in this study from August 2007 to May 2008 in Affiliated Hospital of Nantong University, Jiangsu. Sixty-five healthy volunteers served as the control. The fluorescence of the PCR products was detected continuously during the amplification by RFQ-PCR. According to the standard curves created by plasmid DNA, the expression level of target genes in clinical samples was determined using software. The results were presented as the ratios of target genes to beta(2)-microglobulin (beta(2)-M) mRNA, and compared with those obtained by conventional cytological method. RESULTS: The detection range of the assay was from 38 copies/microl to 3.8 x 10(6) copies/microl. The coefficients of variation values of both intra-experimental and inter-experimental reproducibility were 8.5% and 13.6%, respectively. The expression of APRIL mRNA in tumor sputum was higher than that in benign pulmonary disease and healthy volunteers (t = 10.50, 11.32, P < 0.01). The positive rate for APRIL mRNA expression was 81.7% (58 of 71) in sputum samples of NSCLC, 3.2% (2/62) in benign pulmonary disease and 1.5% (1/65) in healthy volunteers when cut-off values for positivity were set at the x(-) +/- 2 s of mRNA expression in health volunteers. The level of APRIL mRNA of NSCLC was not related to sex, age, smoking status, TNM stage and lymph node metastasis (P > 0.05, respectively), but was related to pathology subtype and the location of tumors (P < 0.05, respectively). The APRIL mRNA assay (82%) produced a higher detection rate than conventional cytological method (14%) (chi(2) = 67.68, P < 0.01). CONCLUSION: Measurement of the expression of APRIL mRNA in sputum by RFQ-PCR showed high sensitivity and specificity, which maybe useful in diagnosing NSCLC.
