Objective To investigate the effects of recombinant adenovirus-mediated co-transfection of carcinoembryonic antigen (CEA) gene and erythropoietin (EPO) gene on promoting hematopoietic stem cells directly producing erythrocyte vaccine against colon cancer. Methods The expression adenovirus vectors carrying CEA and EPO or green fluorescent protein (GFP) gene were constructed respectively, and recombinant adenovirus carrying CEA, EPO or GFP were packaged and produced respectively. The bone marrow-derived mesenchymal stem cells (MSCs) of mice were isolated and cultured in vitro by anti-CD117 magnetic bead separation, and were transfected with CEA (CEA group), EPO (EPO group) or GFP (blank vector group), co-transfected with CEA and EPO (CEA-EPO group). The expressionsof CEA and EPO gene and its protein after transfection in supernatant fluid of culture were detected by realtime-PCR and Western blot method in each group. We had checked and obtained the vaccine with co-transfection of CEA gene and EPO gene by cell red line marker antibody CD71 and GPA, then we carried on experiments with the vaccine in vitro and in vivo. There were 4 groups in our trail: blank vector group, CEA group, EPO group, and CEA-EPO group. Results We had successfully gathered the hematopoietic stem cells, flow cytometry analysis result showed that there were significant differences before and after purification for positive selected samples (P<0.05). The expressions of double genes (CEA-EPO gene) and protein showed CEA-EPO gene were successfully transfected into the hematopoietic stem cells. We had confirmed erythrocyte vaccine with co-transfection of CEA and EPO gene by antibody CD71 and GPA with flow cytometry. The monocytes cytotoxicity on colon cancer cell line CT26 showed that lysis of target cells of CEA-EPO group were higher than those of other 3 groups when in proportion of 40∶1 (P<0.05). In the experimentation of neoplasma format, the volume of tumor and mortality were smaller or lower, but survival time was longer of CEA-EPO group in2 weeks after treatment (P<0.05). Conclusions The erythrocyte vaccine with co-transfection of CEA gene and EPO gene has efficient anti-tumor effects on colon cancer. Not only can promote hematopoietic stem cell directly producing erythrocyte vaccine, but also can produce tumor antigen vaccine against colon cancer.
Objective To observe the expression of Ezrin protein in primary carcinoma of gallbladder tissue and the levels of CEA and CA19-9 in serum of patients with primary carcinoma of gallbladder, and to explore the relationship between the expressions of these measurements and clinicopathologic characteristics. Methods Immunohistochemistry was applied to analyze the expression of Ezrin protein in primary carcinoma of gallbladder and chronic cholecystitis tissue. The levels of CEA and CA19-9 in serum and clinicopathologic characteristics of all including patients were detected with clinical measurement. All data were analyzed statistically. Results ①The positive rates of Ezrin protein in primary carcinoma of gallbladder and chronic cholecystitis tissue were 66.7% （40/60） and 30.8%（4/13）, respectively (χ2=5.57, Plt;0.05). ②There was no difference between the expression of Ezrin protein in primary carcinoma of gallbladder tissue and age or gender (Pgt;0.05). However, difference was significant between the Ezrin expression and degree of difference, pNevin stages, pTNM stages, lymph node metastasis or distant metastasis (Plt;0.05). ③There were no differences between the positive rates of CEA and CA19-9 in primary carcinoma of gallbladder and age or gender (Pgt;0.05). However, differences were significant between the positive rates of CEA and CA19-9 and pNevin stages, pTNM stages, degree of difference, lymph node metastasis or distant metastasis (Plt;0.05). ④There was some relationship between the expression of Ezrin protein and the positive rate of CEA (rs=0.213, Plt;0.05), but not with the positive rate of CA19-9 (rs=0.081, Pgt;0.05). Conclusions The high expression of Ezrin protein may promote the invasion and metastasis in primary carcinoma of gallbladder. It could be possible to decide the outcome of primary carcinoma of gallbladder through the combined analysis on the expression of Ezrin protein and the serum levels of CEA and CA19-9.
【Abstract】ObjectiveTo detect the spreading scope of rectal cancer to mesorectum by RT-PCR using carcinoembryonic antigen (CEA) mRNA as a marker and to investigate the excision scope of mesorectum in resection of rectal cancer. MethodsForty specimens from 40 rectal cancer patients who underwent curative operation was employed to detect the metastatic deposits scattered in the mesorectum by RT-PCR using CEA as a marker. ResultsNine of 40 (22.5%) specimens contained metastatic deposits scattered in the mesorectum. The metastasis was just within the range of 4cm mesorectum under the verge of tumor. The tumor spreading to mesorectum is correlated with Dukes stages，the infiltrated depth of bowel wall, tumor differentiation and tumor type(P＜0.05), and is not correlated with the size of tumor and the level of CEA(Pgt;0.05). ConclusionThe excision of mesorectum should be within the range of 5cm under the verge of tumor in surgical management of rectal cancer.
Objective To evaluate the value of bile and serum CA19-9 in diagnosing biliary tract carcinoma. Methods Bile and serum CA199 and CEA were determined by radioimmunoassay (RIA). Results The dividing value of bile CA199 is 12 000 kU/L, and its sensitivity and specificity were 85.71%, 73.91% respectively. The dividing value of bile CEA is 480 μg/L, and its corresponding indexes were 57.14% and 77.17%. The false positive rate of bile CA19-9 and CEA were 26.09% and 22.83%. Serum CA19-9 sensitivity, specificity were 80.00% and 85.11%; the corresponding indexes of serum CEA were 68.57% and 82.97%. Conclusion CA19-9 is an effective tumor marker in diagnosing, deciding whether the tumor has been radically resected and in monitoring its response to the treatment.
【Abstract】Objective To compare the reliability of serum tumour specific growth factor (TSGF) with carcinoembryonic antigen (CEA) in the diagnosis of tumour. Methods The patients were divided into two groups according to malignancy and benignity. In benignity, the patients were subdivided into inflammatory and non-inflammatory groups. The levels of TSGF and CEA in the two groups were measured. Results The positive rate of TSGF and CEA in malignant group was 67.41% and 38.84% respectively; that in benign was 24.56% and 2.63% respectively, in which the inflammatory group was 32.35% and 5.88% respectively, and in non-inflammatory group was 18.25% and 0% respectively. The positive rate of TSGF and CEA was higher in malignant than in benign group (P＜0.005). The positive rate of TSGF was higher than CEA in malignant (P＜0.005) and inflammatory group (P＜0.005). Conclusion Serum TSGF is a useful blood marker in the diagnosis of patients with malignancy, and is a more sensitive and broad-spectrum marker than CEA for the diagnosis of tumours. CEA is more specific than TSGF for the diagnosis of tumours. Combined measurement both TSGF and CEA will enhance the diagnostic rate.
Objective To study effect of carcinoembryonic antigen (CEA) positive targeted lymphocytes on gastric cancer cells in vitro and in vivo. Methods The peripheral blood mononuclear cells (PBMCs) were isolated from the peripheral blood of healthy volunteers. The recombinant vector anti-CEA-scFv-CD3ζ-pcDNA3.0 was transfected into the PBMCs by lipofectamine 2000, by this means, the CEA special lymphocytes were obtained. Meanwhile, the PBMCs transfected with empty plasmid pcDNA3.0 were used as control (empty vector lymphocytes). The different lymphocytes and gastric cancer cells (CEA positive KATOⅢ gastric cancer cells and CEA negative BGC-823 gastric cancer cells) were co-cultured, then the ability to identify the gastric cancer cells and it’s effect on apoptosis of gastric cancer cells were observed at 24 h or 36 h later respectively. The CEA special lymphocytes and empty vector lymphocytes were injected by the tail vein of nude mice bearing gastric cancer cells, then it’s effect on the tumor was observed. Results ① The CEA special lymphocytes could strongly identify the KATOⅢ gastric cancer cells (identification rate was 72.3%), which could weakly identify the BGC-823 gastric cancer cells (identification rate was 7.8%). ② The apoptosis rate of the co-culture of CEA special lymphocytes and KATOⅢ gastric cancer cells was significantly higher than that of the co-culture of empty vector lymphocytes and KATOⅢ gastric cancer cells (P=0.032), which had no significant difference between the co-culture of CEA special lymphocytes and BGC-823 gastric cancer cells and the co-culture of empty vector lymphocytes and BGC-823 gastric cancer cells (P=0.118). ③ The tumor volume of the co-culture of CEA special lymphocytes and KATOⅢ gastric cancer cells was significantly smaller than that of the co-culture of empty vector lymphocytes and KATOⅢ gastric cancer cells (F=5.010, P<0.01) or the co-culture of CEA special lymphocytes and BGC-823 gastric cancer cells (F=4.982, P<0.01), which had no significant difference between the co-culture of CEA special lymphocytes and BGC-823 gastric cancer cells and the co-culture of empty vector lymphocytes and BGC-823 gastric cancer cells (F=1.210, P>0.05). Conclusion CEA special lymphocytes can promote cell apoptosis and inhabit tumor reproduction of CEA positive gastric cancer cells in vitro and in vivo.
Objective To evaluate branched-chain DNA (b-DNA) signal amplification and semi-quantitative (Sq) RT-PCR in detection of free cancer cells in peritoneal flushing fluid of colorectal cancer patients during surgery. Methods The CEA mRNA in peritoneal flushing fluid in 48 cases of colorectal cancer were detected by b-DNA and SqRT-PCR. Peritoneal flushing fluid cytology (PLC) was conformed simultaneously to detect the free cancer cells. The peritoneal flushing fluid of 12 cases with colorectal benign disease were taken as negative control, GAPDH mRNA as internal control. Results In colorectal cancer patients, positive rate of free cancer cells by bDNA and SqRT-PCR （43.8%, 31.3%） was higher than that by PLC （4.2%）. The relative quantitative expressions of CEA mRNA were related to the Dukes staging， depth invasion and differentiation degree (Plt;0.05)， but irrelevant to tumor size，the patients’ age and gender (Pgt;0.05).Conclusion Both b-DNA and SqRT-PCR technologies have advantages and disadvantages to detect free cancer cells in peritoneal flushing fluid， which are related to clinicopathological factors.
Carcinoembryonic antigen (CEA)was measured with ABC immunohistochemistry method in fourty-one gastric cancer tissues and sixty-six tissue from normal stomach and gastric benign lesions. The study revealed that the reactive signals in the former were ber than those in the latter. Simultaneously, CEA localized mainly in the cytoplasm or stroma in the cancerous tissue, but in normal gastric tissue or benign gastric lession, CEA distributed mainly in the margin of gland with gastric depression or membranous type. The result also revealed that the distribution patterns of ECA were linked with the cell growth types and infiltrating of gastric cancer. The authors consider that the expression state of CEA in gastric cancer is correlated with its biological behavior, and distribution patterns of CEA are more clinically significant than reactive intensities in the tissue. Patients have different prognosis with different CEA distribution patterns in tissue though their pathological types and TNM stages are the same.
Objective To Evaluation of Accuracy and Quality of Diagnostic Test of CEA for the Diagnosis of NSCLC in Chinese Patients. Methods We searched Chinese Biological Medicine Database (CBM, 1978 to 2009) and China National Knowledge Infrastructure (CNKI, 1994 to 2009). Diagnostic tests of CEA for the diagnosis of NSCLC were included. Data were extracted, and the quality of included studies was evaluated according to the six criteria of diagnostic tests. The heterogeneity test and The Summary Receiver Operating Characteristic (SROC) curve and meta-analyses were performed by MetaDisc. Results A total of 84 relevant articles were retrieved and 11 were included in our review. Eleven studies involving 925 patients (861 NSCLC patients, all diagnosed by the gold standard) were included. Meta-analyses showed that the heterogeneity among studies was high (P=0.000 2, I2=69.1%), the pooled sensitivity was 0.542 and the pooled specificity was 0.869. Subgroup analyses indicated that 5 of the studies which used the ECLIA (P=0.376, I2=5.4%, AUC= 0.748 3) and 4 of the studies which lung adenocarcinoma (P=0.186, I2=37.6%, AUC=0.900 2) and 4 of the studies which lung squamous cell carcinoma (P=0.955，I2=0.00%, AUC=0.762 0) had no heterogeneity. serum CEA is low sensitive and high specific on the diagnosis of NSCLC. The sensitivity and diagnostic accuracy rate of CEA were higher in adenocarcionoma than squamous cell cance. Conclusion CEA could be regarded as one of the reference tests in patients with NSCLC, Serum CEA is more sensitive and specific than lung squamous cell carcinoma on lung adenocarcinoma. but more high quality trials are required.