Objective To verify the expressions of genes associated with colorectal cancer with hyperglycemia and evaluate their diagnostic values.Methods Tumor tissues,distal normal intestinal mucosa,and peripheral blood samples were harvested from 109 colorectal cancer patients and peripheral blood samples from 30 diabetes patients and 30 healthy volunteers. The mRNA expressions of glucose regulated protein 78 (GRP78),NADPH oxidase-1 (NOX1),carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5),heat shock protein 60 (HSP60),and histone deacetylase 1(HDAC1) were detected by real-time quantitative polymerase chain reaction. The correlation between the gene expressions and clinicopathological parameters in colorectal cancer patients were analyzed using Pearson's correlation analysis. Diagnostic test accuracy evaluation was used to calculate the sensitivity,specificity,accuracy,predictability,Youden index,and likelihood ratio of serum gene expressions in colorectal cancer patients,and the receiver operating characteristic (ROC) curves were drawn. The area under the ROC curve was calculated to evaluate the diagnostic efficiency of the combined detection of multiple genes.Results The mRNA levels of GRP78 (P=0.001),NOX1 (P=0.022),CEACAM5 (P=0.000),HSP60 (P=0.044),and HDAC1 (P=0.047) were positively correlated with the fasting blood glucose level. The mRNA expressions of NOX1 (P=0.000,P=0.008) and HDAC1 (P=0.000,P=0.037) in tissues and serum were significantly higher in colorectal cancer patients than in patients with normal blood glucose levels. The NOX1 mRNA expression was positively correlated with the diameter of colorectal cancer (P=0.013),and the HDAC1 mRNA expression was significantly correlated with the tumor site (P=0.049),depth of primary tumor invasion (P=0.025),and TNM stage (P=0.042). The areas under the ROC curves of NOX1,CEACAM5,and HDAC1 were 0.931,0.852,and 0.860 respectively (all P=0.000). The specificity,accuracy,and negative predictive value of NOX1,HDAC1 mRNA expression in colorectal cancer patients with hyperglycemia were all above 90%. The diagnostic sensitivity and specificity of the combined detection of NOX1,CEACAM5,and HDAC1 were 98.82% and 99.93%,respectively.Conclusion Combined detection of genes associated with colorectal cancer accompanied by hyperglycemia can improve the diagnostic efficiency of early screening.
Biomarkers, Tumor ; genetics ; Carcinoembryonic Antigen ; genetics ; Case-Control Studies ; Colorectal Neoplasms ; complications ; diagnosis ; genetics ; Diabetes Mellitus ; genetics ; GPI-Linked Proteins ; genetics ; Heat-Shock Proteins ; genetics ; Histone Deacetylase 1 ; genetics ; Humans ; Hyperglycemia ; complications ; diagnosis ; genetics ; NADPH Oxidase 1 ; genetics ; ROC Curve

The 8th edition of AJCC cancer staging system will be launched all over the world in January 1, 2018. The major advances in the 8th edition are the introduction of non-anatomic prognostic and predictive factors supported by I(-II( grade evidence based on histopathology and molecular biology, and the improvement of prognostic assessment system based on these factors, including CEA level, cancer retraction score, circumference margin, lymphatic invasion, peripheral nerve invasion, microsatellite instability, KRAS/NRAS gene mutation and BRAF gene mutation. In the background of evidence-based medicine and precise medicine, combination of anatomic staging and non-anatomic classification system is very important for establishing and improving the prognostic and predictive assessment system of colorectal cancer. This will help to assess colorectal cancer staging and grouping much better, evaluate the prognosis, predict individualized efficacy, and promote clinical practice of colorectal cancer from traditional population-based diagnosis and treatment to the precise individualized care.
Carcinoembryonic Antigen ; blood ; Colorectal Neoplasms ; classification ; diagnosis ; Female ; Humans ; Male ; Microsatellite Instability ; Mutation ; Neoplasm Invasiveness ; Neoplasm Staging ; standards ; Precision Medicine ; methods ; trends ; Prognosis ; Proto-Oncogene Proteins B-raf ; genetics ; Proto-Oncogene Proteins p21(ras) ; genetics

Lipooligosacharide (LOS) of Neisseria gonorrhoeae (gonococci, GC) is involved in the interaction of GC with host cells. Deletion of the alpha-oligosaccharide (alpha-OS) moiety of LOS (lgtF mutant) significantly impairs invasion of GC into epithelial cell lines. GC opacity (Opa) proteins, such as OpaI, mediate phagocytosis and stimulate chemiluminescence responses in neutrophils in part through interaction with members of the carcinoembryonic antigen (CEA) family, which includes CEACAM3 (CD66d), a human neutrophil specific receptor for phagocytosis of bacteria. In the present work, we examined the effects of OpaI-expressing lgtF mutant on phagocytosis by HeLa-CEACAM3 cells and chemiluminescence responses in neutrophils. The results showed that lgtF mutant even expressing OpaI completely lost the ability to promote either phagocytosis mediated by CEACAM3 interaction in HeLa cells or chemiluminescence responses in neutrophils. These data indicated that Opa proteins in the lgtF mutant, which might result from the conformational change, cannot be functional.
Antigens, Bacterial ; chemistry ; genetics ; immunology ; metabolism ; Carbohydrate Sequence ; Carcinoembryonic Antigen ; genetics ; immunology ; Gene Expression Regulation ; HeLa Cells ; Host-Pathogen Interactions ; Humans ; Lipopolysaccharides ; chemistry ; immunology ; Luminescent Measurements ; Mutation ; Neisseria gonorrhoeae ; genetics ; metabolism ; pathogenicity ; Neutrophils ; immunology ; microbiology ; Phagocytosis

BACKGROUNDColorectal carcinoma is one of the most common malignant tumors. Despite advances in therapy, mortality is still very high. The aim of this study was to evaluate the expression of paxillin in the human colon adenocarcinoma cell line SW480 and its role in cell cycle and apoptosis. We also investigated the expression of paxillin in colorectal carcinoma tissues and its relationship to clinicopathological features and survival.

METHODSPaxillin short hairpin RNA (shRNA) was constructed and transfected into the colon adenocarcinoma cell line SW480. The influence of paxillin shRNA on the cell cycle and cell apoptosis was analyzed by flow cytometry. Immunohistochemistry staining was used to assess the expression of paxillin and its association with the expression of carcinoembryonic antigen (CEA), carbohydrate antigen (CA) 19-9, p53 and Bcl-2 in 102 patients with primary colorectal carcinoma. Western blotting was also used to investigate the expression of paxillin. Medical records were reviewed and a clinicopathological analysis was performed.

RESULTSIn vitro, the percentage of cells in S phase was (45.23±1.05)%, (43.53±1.23)%, and (36.13±0.57)% in the blank control group, negative control group, and paxillin shRNA group respectively. It was significantly decreased in the paxillin shRNA group (P = 0.000). The early apoptosis index of the paxillin shRNA group (17.2±1.18%) was significantly increased compared to the control shRNA group ((13.17±1.15)%, P = 0.013). Paxillin was positive in 71 (69.6%) patients, and it was found to be overexpressed in tumor tissues compared with normal adjacent tissues. Paxillin positive rate was higher in patients who are less than 50-years old (100.0% vs. 65.6%, P = 0.016). Paxillin expression was associated with a high histologic grade of carcinoma (81.4% vs. 61.0%, P = 0.031), a high rate of regional lymph node metastasis (22.5% vs. 13.0%, P = 0.031), mesenteric artery lymph node metastasis (100.0% vs. 64.8%, P = 0.008), distant metastasis (94.1% vs. 64.7%, P = 0.016) and a high Tumor Node Metastasis (TNM) stage (94.1%, 73.2%, 60.0%, and 50%, P = 0.030). Multivariate analyses revealed that recurrence was associated with the rate of regional lymph node metastasis (P = 0.001) and paxillin expression (P = 0.024). Multivariate analysis indicated that the overall survival is related to the TNM stage (P = 0.000).

CONCLUSIONSIn vitro, paxillin may promote cell proliferation and inhibit apoptosis in SW480 cells. Paxillin may be a potential metastasis predictor, and an independent prognosis factor of recurrence. It may also be related to poor patient outcomes, but was not an independent predictor of survival.

Apoptosis ; genetics ; physiology ; Biomarkers, Tumor ; genetics ; metabolism ; Carcinoembryonic Antigen ; metabolism ; Cell Cycle ; genetics ; physiology ; Cell Line, Tumor ; Cell Proliferation ; Colorectal Neoplasms ; genetics ; metabolism ; Female ; Humans ; Immunohistochemistry ; In Vitro Techniques ; Male ; Paxillin ; genetics ; metabolism ; RNA, Small Interfering ; genetics

OBJECTIVETo study the feasibility of preparing a therapeutic lung cancer vaccine by transfecting dendritic cells (DCs) with adeno-associated virus vector carrying carcino-embryonic antigen gene (rAAV/CEA).

METHODSAdherent cells (monocytes) isolated from the peripheral blood of a healthy donor were infected with rAAV/CEA virus stock or pulsed with CEA peptide (control). The monocytes in both groups were induced into mature DCs with recombinant human GM-CSF, IL-4 and TNF-α. At day 7 of induction, the mature DCs were harvested and mixed with T lymphocytes. T cell proliferation stimulated by the DCs was assessed with (3)H-thymidine uptake, and the expression of IL-4, IFN-γ, CD8, CD4, CD25 and CD69 in cytotoxic T lymphocytes (CTL) was analyzed with flow cytometry. The cytotoxicity of the CTL against the target CEA-positive lung cancer A549 cells was tested by (51)Cr releasing assay.

RESULTSThe DCs transfected with rAAV/CEA strongly stimulated the proliferation of the T cell populations, and the induced CTL showed high expressions of CD8, CD69 and IFN-γ. The transfected DCs exhibited a high killing ability of CEA-positive lung cancer cells, and the killing showed a CEA antigen specificity and was limited by MHC I. These results suggested the ability of rAAV/CEA-transfected DCs in generating specific cellular immunity in vitro.

CONCLUSIONIt is feasible to prepare therapeutic lung cancer vaccines by transfecting DCs with rAAV/CEA.

Cancer Vaccines ; Carcinoembryonic Antigen ; genetics ; Cell Line ; Dendritic Cells ; immunology ; Dependovirus ; genetics ; Genetic Vectors ; Humans ; Monocytes ; immunology ; Transfection

Chronic inflammation is thought to be the leading cause of colorectal cancer, and interleukin-10 (IL10) has been identified as a potent immunomodulatory cytokine that regulates inflammatory responses in the gastrointestinal tract. Although several single nucleotide polymorphisms (SNPs) in IL10 have been associated with the risk of colorectal cancer, their prognostic significance has not been determined. Two hundred and eighty-two colorectal cancer patients were genotyped for two candidate cancer-associated SNPs in IL10. The associations of these SNPs with distant metastasis-free survival and overall survival were evaluated by Kaplan-Meier analysis and Cox regression model. The minor homozygote GG genotype of IL10 rs3021094 was significantly associated with a 3.30-fold higher risk of death compared with the TT+TG genotypes (P=0.011). The patients with IL10 rs3021094 GG genotype also had a poorer overall survival in Kaplan-Meier analysis (log-rank P=0.007) and in multivariate Cox regression model (P=0.044) adjusting for age, gender, carcinoembryonic antigen levels, tumor differentiation, stage, lymphovascular invasion, and perineural invasion. In conclusion, our results suggest that IL10 rs3021094 might be a valuable prognostic biomarker for colorectal cancer patients.
Aged ; Alleles ; Carcinoembryonic Antigen/blood ; Cell Differentiation ; Colorectal Neoplasms/*genetics/mortality/pathology ; Female ; Genotype ; Homozygote ; Humans ; Interleukin-10/*genetics ; Kaplan-Meier Estimate ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; *Polymorphism, Single Nucleotide ; Regression Analysis ; Tumor Markers, Biological/genetics

OBJECTIVETo establish a colon cancer cell line with stable expression of carcinoembryonic antigen(CEA).

METHODSRecombinant lentivirus conjugated with CEACAM5 cDNA were used to transfect wild CT26 cells. Antibiotics were given for 2 weeks to select CEA positive cells. A single transfected clone was obtained using limiting dilution. The 7th and 14th passages of cells cultured in vitro were detected for CEACAM5 mRNA by RT-PCR, and protein by western blot. The location of CEACAM5 expression was examined using fluorescent microscope and immunocytochemistry. The 14th passage cells were injected subcutaneously into mice to create BALB/c model and CEACAM5 protein was detected by in vivo fluorescence image analysis system and immunohistochemistry.

RESULTSCEACAM5 mRNA and protein were found in the 7th and 14th passages of CT26CEA cells, which were proved to locate in the cytoplasm by fluorescence microscope and immunohistochemistry. Abundant CEACAM5 protein was found in subcutaneous tumors by in vivo fluorescence image analysis system and immunohistochemistry.

CONCLUSIONColon cancer cell line CT26 with stable expression of CEA in vitro and in mice can be used as a suitable tool to facilitate research on the impact and mechanism of CEA on colon cancer under normal immune environment.

Animals ; Carcinoembryonic Antigen ; genetics ; metabolism ; Cell Line, Tumor ; Colonic Neoplasms ; genetics ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Neoplasm Transplantation ; RNA, Messenger ; genetics ; Transfection

OBJECTIVETo study the effect of CEA gene regulation on the anti-tumor activity of oncolytic adenovirus H101 to esophageal carcinoma, and to explore the intrinsic factors influencing H101 sensitivity.

METHODSStable human esophageal cancer cell line EC9706 cells with lower (EC9706-SCEA) and higher CEA expression (EC9706-CEA) were chosen, thawed and cultured, and then to analyse the influence of CEA expressed at different levels on cell growth. The cytotoxic effect of H101 was assayed by in vitro and nude mouse in vivo.

RESULTSThe cell growth experiment showed that the population doubling time of EC9706-SCEA, EC9706-CEA and EC9706 cells were (30.9 ± 2.0) h, (31.1 ± 2.5) h and (29.1 ± 2.6) h, respectively, showing no significant difference among them (P > 0.05). The cytotoxic activity of H101 was higher on EC9706-SCEA than on other four groups, when MOI was ≥ 0.01 PFU (P < 0.05). The mouse experiment showed that H101 inhibited the growth of transplanted tumors in all experimental groups. Its effect on CEA-silenced tumors (inhibition rate was 61.5% to 74.5%) was significantly higher than that on CEA-overexpression tumors (32.3% to 38.5%) and control EC9706 transplanted tumors (35.5% to 44.8%). There was a significant difference between them (P < 0.05).

CONCLUSIONSThe results in vitro and in vivo experiments show that H101 can enhance the cytotoxic effect on EC9706 cells with lower CEA expression. To silence the expression of CEA may provide a novel strategy for target gene therapy of esophageal carcinoma.

Adenoviridae ; physiology ; Animals ; Carcinoembryonic Antigen ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Esophageal Neoplasms ; metabolism ; pathology ; therapy ; Female ; Gene Silencing ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Oncolytic Virotherapy ; Oncolytic Viruses ; physiology ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Tumor Burden

OBJECTIVETo investigate the expression of cyclooxygenase-2(COX-2) and CEA in the tissues adjacent to the tumor within different distances.

METHODSA total of 42 colorectal cancer tissues were collected.The adjacent tissues within 3 cm to the tumor were procured every 1 cm. Normal tissue was also collected. RNA was extracted and the expression of CEA and COX-2 was detected by RT-PCR.

RESULTSThe CEA mRNA levels of the tumor, the tissues of every 1 cm adjacent to the tumor, and the normal tissue were 135.2 ± 23.3, 78.2 ± 17.3, 75.9 ± 16.5, 56.2 ± 10.7, 52.3 ± 12.8, 18.2 ± 7.9, 16.2 ± 6.5, and 16.6 ± 7.0. The levels of COX-2 mRNA in above positions were 134.9 ± 31.1, 79.2 ± 20.2, 77.0 ± 20.5, 62.7 ± 21.9, 58.0 ± 18.1, 21.2 ± 10.3, 18.3 ± 7.6, and 17.1 ± 6.3. These data showed a decreasing trend of CEA and COX-2 as the distance increased from the tumor. The CEA mRNA levels showed positive correlation with the levels of COX-2 mRNA(r=0.725, P<0.01).

CONCLUSIONCEA and COX-2 may be considered to be used as biomarkers for the study of molecular resection margin of colorectal cancer.

Adult ; Aged ; Carcinoembryonic Antigen ; metabolism ; Colorectal Neoplasms ; genetics ; immunology ; pathology ; Cyclooxygenase 2 ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Staging

OBJECTIVETo assess the value of serum carcinoembryonec antigen (CEA) in monitoring the response to biochemotherapy by Herceptin plus taxol (TAX) in patients with Her-2-positive advanced breast cancer.

METHODSThe changes in serum CEA level were investigated retrospectively after two cycles of biochemotherapy in 83 patients with Her-2-positive advanced breast cancer. The correlations between the changes and radiological objective response were analyzed.

RESULTSAfter two cycles of biochemotherapy, the clinical benefit rate (CBR) was 81.9%. In the 60 patients with lowered CEA level, the CBR was 85.0% (51/60), with a non-response rate of 15.0% (9/60); in contrast, the CBR was only 34.8% in 23 patients with elevated CEA, with a non-response rate of 65.2%, showing significant difference between the two groups (P<0.05).

CONCLUSIONSerum CEA level can be used to monitor the therapeutic effect of biochemotherapy in patients with Her-2-positive advanced breast cancer.

Adult ; Aged ; Antibodies, Monoclonal, Humanized ; administration & dosage ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Breast Neoplasms ; drug therapy ; genetics ; Carcinoembryonic Antigen ; blood ; Female ; Humans ; Middle Aged ; Monitoring, Physiologic ; Paclitaxel ; administration & dosage ; Receptor, ErbB-2 ; genetics ; Trastuzumab