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 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.
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.
ObjectiveTo investigated the levels of aldolase A (ALDOA) in pleural effusion in patients with different pathological types of lung cancer and patients with tuberculous pleurisy,and the correlation between ALDOA and carcinoembryonic antigen (CEA),lactate dehydrogenase(LDH). Methods80 cases of pleural effusion samples were collected,of which 65 cases of lung cancer (malignant group) and 15 cases of tuberculous pleurisy (TB group). All the patients were not treated with anti-inflammatory or steroid therapy. ALDOA concentrations in pleural effusion were detected by ELISA and the contents of CEA and LDH in pleural fluid were detected by chemiluminescence assay. ResultsThe levels of ALDOA,CEA and LDH in the malignant group were 46.75±21.39 ng/mL,82.24±56.63 ng/mL,755.76±382.54 U/L respectively,and were 23.92±17.21 ng/mL,2.55±1.67 ng/mL,and 388.37±163.87 U/L in the TB group respectively. The levels of ALDOA,CEA and LDH in the malignant group were significantly higher than those in the TB group (P<0.01). The concentrations of ALDOA in malignant pleural effusion from different pathological types of lung cancer were 71.65±32.09 ng/mL(adenocarcinoma),22.43±18.23 ng/mL(small cell lung cancer),and 19.16±13.85 ng/mL(squamous cell carcinoma),respectively. The concentration of ALDOA in malignant pleural effusion from the adenocarcinoma patients was significantly higher than that in the other two types (P<0.05). The concentration of CEA was 112.40±62.71 ng/mL(adenocarcinoma),62.45±54.78 ng/mL(small cell lung cancer),and 71.87±52.4 ng/mL(squamous cell carcinoma),respectively. It was significantly higher in adenocarcinoma than that in other two types (P<0.05). The levels of LDH were 661.81±328.93 U/L(adenocarcinoma),737.62±315.41 U/L(small cell lung cancer),767.85±503.28 U/L(squamous cell carcinoma),respectively. There was no significant difference in three types(P>0.05). The concentrations of ALDOA in pleural effusion from the patients with lung cancer or tuberculous pleurisy were positively correlated with the concentrations of CEA and LDH (P<0.01 or 0.05). ConclusionThe levels of ALDOA,CEA and LDH in malignant pleural effusion from lung cancer patients were significantly higher than those in pleural effusion from patients with tuberculous pleurisy. The ALDOA and CEA levels in malignant pleural effusion from lung adenocarcinoma patients were significantly higher than those in small cell lung cancer and squamous cell carcinoma patients. There were highly positive correlation between ALDOA,CEA and LDH levels.
Serum tumor markers CEA, CA19-9, CA72-4 and Helicobacter pylori (H.pylori) antibodies were measured in 162 patients with gastric cancer. CEA, CA19-9 and CA72-4 had sensitivities of 24.0%, 35.5% and 21.9% respectively. CA72-4 provided 100% specifity, compared to 77% and 93% for CA19-9 and CEA. The positive predictive value (PV) in CEA, CA19-9 and CA72-4 was higher than negative PV. Serum CA19-9 and CA72-4 levels rose in tumor of ＞5.0cm in diameter. The CA19-9 increased remarkably when the deeper stomach wall was invased. The significantly elevated CEA, CA72-4 and CA19-9 levels were found in patients who had nodal involvement in more than 50% and distant metastasis. However, the increase of CEA, CA19-9 and CA72-4 were found in undifferentiated tumor. Antibodies to H.pylori were detected in 54% of patients but in only 22% control subjects. A significant association was found between H.pylori infection and gastric cancer (odds ratio=3.75; 95% confidence interval=2.11-5.41, Ｐ＜0.01). Conclusions: CEA, CA19-9 and CA72-4 have higher specifity but lower sensitivity in diagnosis of the gastric cancer. The levels of CEA, CA19-9 and CA72-4 are significantly associated with the diameter, the depth of invasion, nodal involvement, distant metastasis and cell differention. Infection with H.pylori may be an important cause of gastric cancer.
ObjectiveTo explore the application value of the combined detection of CA19-9, CA72-4, carcinoembryonic antigen (CEA), serum pepsinogen Ⅰ(PGⅠ), serum pepsinogen Ⅱ(PGⅡ), ratio of PGⅠ and PGⅡ (PGR), and gastrin-17 (G17) in the diagnosis of gastric cancer.MethodsOne hundred cases of gastric cancer admitted to the Joint Logistic Support Force 940 Hospital of the People’s Liberation Army from January 2016 to August 2018 were respectively collected as the observation group, 110 cases of benign gastric lesions as the control group during the same period, the levels of serum CA19-9, CA72-4, CEA, PGⅠ, PGⅡ, PGR, and G17 were tested among patients in the two groups, the diagnostic value of single and combined detection (included CA19-9, CA72-4, CEA, PGⅠ, PGⅡ, PGR, and G17) were explored.ResultsThe levels of CA19-9, CA72-4, CEA, and G17 in the observation group were higher than those of the control group (P<0.05), the levels of PGⅠ and PGR were lower than those of the control group (P<0.05). The positive detection rates of CA19-9, CA72-4, CEA, G17, PGⅠ, PGR, and combined detection in the observation group were all higher than those of the control group (P<0.05). The sensitivity and accuracy of the combined detection in the diagnosis of gastric cancer were higher than that of single serum index (P<0.05). The levels of serum CA19-9, CA72-4, CEA, and G17 in the patients of Ⅲ+Ⅳ period, low and moderate degree of differentiation, the tumor diameter was larger than five centimeters, signet-ring cell carcinoma, and distance metastasis of gastric cancer patients were on the high side compared with Ⅰ+Ⅱ period, high differentiation, the tumor diameter was less than or equal to five centimeters, glandular cancer, and no distance metastasis of gastric cancer patients, as well as the levels of serum PGⅠ and PGR on the low side (P<0.05).ConclusionThe combined detection of CA19-9, CA72-4, CEA, PGⅠ, PGⅡ, PGR, and G17 can effectively improve the diagnose rate of gastric cancer, and they are closely related to the pathological characteristics of gastriccancer patients.
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.
【Abstract】ObjectiveTo eliminate the interference of CEA-related substances in CEA measurement and increase the specificity of CEA in the detection of malignant digestive diseases. MethodsCEA level of peripheral blood and digestive juice (bile, gastric juice) from patients with benign or malignant digestive diseases was measured by ELISA, and semi-dry electrophoretic transfer method of Western blot technique to distinguish CEA and CEA-related substances. ResultsIn malignant diseases, the CEA level of digestive juice was significantly higher than that in the blood, and there was no difference of CEA level in digestive juice and blood in benign diseases. Meanwhile, the CEA level of digestive juice and blood in malignant diseases were significantly higher than that in benign diseases. A specific band (molecular weight about 210×103) was detected in all malignant diseases except four cases whose CEA level was too low (less than 5 μg/L), whereas no one of benign diseases had this specific band no matter how high or low the CEA level was. ConclusionThe specificity of CEA detection in malignant digestive diseases can be improved by using digestive juice as sample and combining with Western blot technique.