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.
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.
Radioimmunoassay was performed to measure carcinoembryonic antigen (CEA) levels in gastric juice before and after operation in 51 gastric cancer patients (group Ⅰ)， 33 patients with gastric benign lesion (group Ⅱ) and 8 patients with malignant lesion in digestive system other than gastric cancer (group Ⅲ). The results showed that preoperative CEA levels of in group Ⅰ were the highest among three groups (Ｐ＜0.01), but no statistic difference was noted in group Ⅱ and group Ⅲ. In group Ⅰ and group Ⅱ, postoperative CEA levels were higer than the preoperative levels. The authors believe that preoperative CEA measurement of gstric juice is an accessory method in diagnosing gastric cancer, nevertheless, there is no diagnostic significence of postoperative measurement in patient undergone partial gastrectomy.
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.
ObjectiveTo systematically review the diagnostic value of the combined test of serum CA153, CA125 and CEA in detection of breast cancer. MethodsClinical diagnostic tests about CA153, CA125 and CEA in patients with breast cancer were retrieved in PubMed, EMbase, CBM, The Cochrane Library (Issue 3, 2014), CNKI, VIP, and WanFang Data from January 1st, 2004 to April 16st, 2014. References of included literature were also retrieved. Literature screening according to the inclusion and exclusion criteria, data extraction and methodological quality assessment were completed by two reviewers independently. Meta-analysis was then conducted using Meta-Disc 1.4 software. ResultsA total of 21 studies involving 4 263 subjects were enrolled. In all studies, the results of meta-analysis indicated that:the DOR, AUC and Q index of the single test (CA153) were 18.71 (95%CI 11.62 to 30.11), 0.858 9, and 0.789 7; while those of the combined test (CA153, CA125 and CEA) were 37.95 (95%CI 21.97 to 65.57), 0.959 1, 0.903 1. Compared with the single test, the diagnostic efficacy of the combined test was higher (Z=3.675, P=0.001 5). ConclusionCompared with the detection of CA153 alone, the combined detection of CA125, CA153 and CEA has higher efficacy and accuracy in the diagnosis of breast cancer.
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.
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.
ObjectiveTo explore the effect of five copies hypoxia-responsive element (5HRE) and carcinoembryonic antigen promoter (CEAp) element, and to explore the inhibition effect of lentiviral vectors targeted Ras association domain family 1 isoform A (RASSF1A) gene on SGC7901 human gastric cancer cells. Methods①Expressions of carcinoembryonic antigen (CEA) mRNA and its protein, and RASSF1A protein in SGC7901, MKN28, and MCF-10A cells were detect by real time-PCR (qRT-PCR), immunocytochemistry, and Western blot, to confirm the experimental and negative control cells.②The recombinant vectors of pGL4.20-5HRE-CEAp-Luc were constructed through molecular cloning technique to transfected SGC7901, MKN28, and MCF-10A cells. Each kind of cell was divided into 2 groups:one of them didn't add CoCl2 (normoxia group), and another group added CoCl2 (hypoxia group). Comparison of the fold of activation was performed.③SGC7901 cells were infected by lentiviral vectors of pLV-5HRE-CEAp-RASSF1A (infection group) and negative virus (negative control group), SGC7901 cells without any treatment as blank control group. Then SGC7901 cells of 3 groups were divided into 2 groups:one of them didn't add CoCl2 (normoxia group), and another group added CoCl2 (hypoxia group). The expression of RASSF1A protein was tested by Western blot, and the growth inhibition rate was confirmed by cell counting kit-8 (CCK-8) assay. Comparisons of expression of RASSF1A protein and growth inhibition rate of each group were performed. Results①Results of qRT-PCR, immunocytochemistry and Western blot showed that, SGC7901 cells showed higher expression of CEA mRNA and positive expression of RASSF1A protein than corresponding index of MKN28 cells and MCF-10A cells (P < 0.05), which were assigned as experimental cells; but MKN28 cells showed lower expression of CEA mRNA and negative expression of RASSF1A protein, which were assigned as negative control cells.②In SGC7901 and MKN28 cells transfected recombinant vectors of pGL4.20-5HRECEAp-Luc, compared with normoxia group in the same kind of cell group, the folds of activation in hypoxia group were higher (P < 0.01), but there was no significant difference between the normoxia group and hypoxia group in MCF-10A cells (P > 0.05). In the condition of with or without CoCl2, compared with SGC7901 cells in the same condition, the folds of activation in MCF-10A and MKN28 cells were both lower (P < 0.05); compared with MKN28 cells, the fold of activation in MCF-10A cells was lower (P < 0.05).③Western blot results showed that, in the condition with and without CoCl2, expressions of RASSF1A protein decreased in SGC7901 cells of blank control group and negative control group; weak expressions of RASSF1A protein was observed in SGC7901 cells of infection group when in condition of without CoCl2, but increased when adding CoCl2. But RASSF1A protein didn't expressed in MKN28 cells of blank control group, negative control group, and infection group, whether adding CoCl2 or not. CCK-8 assay result showed that, in SGC7901 cells, the growth inhibition rate of infection group which added CoCl2 was higher than those of other 5 groups (P < 0.05); in MKN28 cells, the growth inhibition rates of infection group and negative group were all higher than those of blank control group, whether adding CoCl2 or not (P < 0.05), but there was no significant difference among the infection group and negative group, whether adding CoCl2 or not (P > 0.05). ConclusionsA new hypoxia inducible and cea-positive tumor-targeting transcriptional regulatory element of 5HRE-CEAp is established successfully, and lentivirus vector of pLV-5HRE-CEAp-RASSF1A can significant inhibit the growth of SGC7901 cells under hypoxia condition.
Objective To explore the correlations of plasma D-dimer and fibrinogen levels with carcinoembryonic antigen (CEA) and cytokeratin 19 fragment (CYFRA21-1) in patients with non-small cell lung cancer (NSCLC). Methods The clinical data of 196 patients with NSCLC diagnosed for the first time in the Department of Respiratory and Critical Care Medicine, the 416 Hospital of Nuclear Indusry between July 2017 and December 2019 were analyzed retrospectively. There were 57 cases in early stage (stage Ⅰ-Ⅱ), 57 cases in medium stage (stage Ⅲ), and 82 cases in advanced stage (stage Ⅳ) according to TNM staging, 108 cases of adenocarcinoma, 87 cases of squamous cell carcinoma, and 1 case of unclassified type according to pathological classification, and 19 deaths and 177 survivals according to outcome. The levels of D-dimer and fibrinogen were determined by immunoturbidimetry and coagulation method, and the levels of CEA and CFYRA21-1 were determined by electro-chemiluminescence method. The non-normally distributed data were presented as median (lower quartile, upper quartile), and Spearman correlation analyses were performed. Results Among the early, middle and advanced stage patients, the levels of D-dimer [198.00 (133.00, 390.87), 279.00 (170.93, 520.89), 389.00 (196.25, 931.00) μg/L], CEA [3.20 (2.60, 5.17), 13.53 (5.07, 70.63), 15.69 (4.07, 123.46) μg/L], and CFYRA21-1 [4.79 (3.15, 8.84), 8.60 (4.83, 19.32), 7.19 (3.09, 15.05) μg/L] were significantly different (P<0.05); however, there was no statistical difference in the level of fibrinogen among the three stages (P>0.05). The level of CYFRA21-1 in the adenocarcinoma group was lower than that in the squamous cell carcinoma group [(5.39 (2.81, 12.71) vs. 6.86 (4.18, 12.29) μg/L, P<0.05], while there was no statistically significant difference in D-dimer, CEA, or fibrinogen between the two groups (P>0.05). The levels of D-dimer, CEA, and CFYRA21-1 in the death group [1176.00 (382.00, 2848.00), 135.34 (24.85, 403.50), 10.82 (7.41, 23.41) μg/L] were significantly higher than those in the survival group [270.00 (146.00, 481.50), 5.62 (3.05, 26.53), 6.28 (3.37, 12.30) μg/L], and the differences were statistically significant (P<0.01); but there was no statistical difference in the level of fibrinogen between the two groups (P>0.05). D-dimer was positively correlated with CEA and CFYRA21-1 (rs=0.450, 0.291; P<0.001), but fibrinogen was not correlated with CEA or CFYRA21-1 (P>0.05). Conclusion D-dimer was more valuable than fibrinogen in predicting the clinical stage and prognosis of NSCLC.