| BackgroundColorectal cancer is one of the most common carcinomas in the digestive tract. There will be a considerable part of patients who underwent a curative resection are going to encounter the situation of tumor recurrence. The 5-year survival rates ranged between 45%-55%.Cytokertin 20(CK20) is a kind of intermediate filament that distributes original cells of the ectoderm, it participates making up the cytoskeleton. CK20 presents a more restrict specificity of epithelial cells. Cytokertin20 is found in nowhere but cells in intestinal mucosa,duodenum mucosa,gastric mucosa, in gland cells in pylorus, in umbrella cells of the urinary system, in merkel cells of epidermis, et al. Cytokertin20 also continually expresses when metaplasia, malignant, metastasis happens or external cultivation. Carcinoembryonic antigen (CEA) is a kind of significant tumor-associated antigen and tumor marker, that is internationally recognized. It is a embryos of oncoprotein weights 180,000 D. CEA expresses widely in epithelial carcinoma, and expresses highly in 90%gastrointesinal cancer,50%breast cancer and 70%non-small cell lung cancer.Objective To investigate the expression and relationships of CEA and CK20 in different tissues of colorectal carcinoma preliminarily, and to investigate the relationship between CEA,CK20 expression and clinical characteristics of patients of colorectal cancer.MethodsDetect the expression of CEA and CK20 respectively using ways of immunohistochemistry in cancer tissue, adjacent normal mucosa, normal mucosa and mesenteric lymph node tissue.ResultsThe expression of CEA and CK20 locates in cytoplasm and membrane of epithelial cells. There is a varying degrees expression of CEA in all the cancer tissue, adjacent normal mucosa, and mesenteric lymph node tissue, however, in normal mucosa, as control group, CEA expresses negatively. The total positive rate of CEA expression in cancer tissue is 92.86%; CEA expresses higher in adjacent normal mucosa than mesenteric lymph node tissue. CK20 expresses verily in every group including in normal mucosa, CK20 expresses higher in cancer tissue than in adjacent normal mucosa, expresses higher in adjacent normal mucosa than in normal mucosa, the mesenteric lymph node tissue has a lowest CK20 expression.Conclusion1,CEA expresses in varying degrees in different colorectal cancer tissue, but not in normal mucosa. There is a certain relationship between Dukes stage and CEA expression in cancer tissue.2,CK20 expresses in varying degrees in all groups including normal mucosa and different colorectal cancer tissue, and the degree of CK20 expression concerns lymph node metastasis status. BackgroundThe metastasis and recurrence of colorectal cancer is closely related to occult micro metastases and circulating tumor cells (CTCS). Such as immunocytochemistry,reverse transcription polymerase chain reaction, flow cytometry, et al. which are usually used for detecting CTCS. However, those methods are either time-consuming or complicated or even result in a false positive outcome, so they are hard to be used in clinical work extensively. There are some new methods have emerged already, for example:real-time fluorescence quantitative PCR, protein fingerprinting technology, nevertheless, these technologies still can't meet the request in daily clinical work because of they are too costly. Immunosensor analysis is applied more and more extensively in recent years, because it's sensitivity, selectivity and simple. Immunosensor is a kind of highly selective biosensor that based on the detection of antigen-antibody binding. Our experiment applied Immunosensor analysis to detect expression of CEA and CK20 in serum of patients of colorectal cancer for the first time.ObjectiveTo investigate a determine system for judging the status and predicting the prognosis of colorectal cancer.MethodsUse ICC,ELISA,real-time fluorescence quantitative PCR and immunosensor analysis to detect the expression of CEA and CK20 in colorectal cancer, respectively, to determine the value of immunosensor analysis in judging the status and predicting the prognosis of colorectal cancer.ResultsUse real-time fluorescence quantitative PCR,immunosensor analysis and ELISA to detect CEA,CK20 in patients'peripheral blood, and to detect CEAmRNA,CK20mRNA in different tissue of colorectal cancer shows that real-time fluorescence quantitative PCR is more sensitive and helpful in improving the positive rate of genetic testing:the positive rate of CEAmRNA in peripheral blood is raised from 57.14% to 89.28%. In 15 patients who are in Dukes A,B,3 patients(741ymph notes) were found to have micro-metastasis through testing CK20 using immunohistochemistry way, testing CK20mRNA copies found 9 more patients(1551ymph notes) to have micro-metastasis. Although the sensitivity and accuracy is not as good as realtime-PCR, the using of immunosensor of CEA and CK20 in detecting clinical specimens results in an outcome that has no significant differences compared with ELISA way, but immunohistochemistry is more specific than realtime-PCR, and results in a higher positive rate than ELISA. Immunosensor of CEA and CK20 can be used to detect CEA quantitatively in serum.Conclusion1,Improving way of detecting CEA,CK20 in patients'peripheral blood of colorectal cancer can achieve a higher positive rate, and going to help effectively diagnosing colorectal cancer and predicting it's prognosis.2,There is great value for realtime-PCR in testing the expression of CK20mRNA of mesenteric lymph nodes in judging the exact status and in predicting the prognosis of colorectal cancer.3,The Embedding method in situ chemical oxidation of dopamine method is established successfully for CEA and CK20 immunosensor, and for testing CEA,CK20 in patients'peripheral blood of colorectal cancer. BackgroundThe most important indexes which conclude the prognosis of colorectal cancer are the infiltrating depth of the tumor and the lymph notes metastasis degree. However, in some colorectal cancer patients who are treated radical surgery with no lymph notes metastasis by pathology routine examination, metastasis and recurrence happen. It is studied that it is related to lymph nodes micro-metastasis of colorectal cancer.In 1992, UICC define occult metastasis and micro-metastasis (MM) as the diameter of single metastatic tumor cell or metastatic tumor cell clump less than 2mm. Major literatures and reports define lymph nodes metastasis focus which are discovered by immune histochemistry instead of routine histological examination. Thus, there is no unified standard in defining lymph nodes micro-metastasis.The former two parts of experiment in this research have proved we can find out occult metastasis and micro-metastasis by detecting CEA,lymph notes of colorectal cancer patients.ObjectiveTo investigate what is the relationship between CEA,CK20 and CEAmRNA,CK20mRNA, and if they can be used as independent factors in deciding status and prognosis of colorectal cancer of assessment system.MethodsBy clinical follow-up and clinical application of statistical methods to compare the two groups of measured values.ResultsPrognostic indicators include:The positive rate of CK20 in peripheral blood, lymph node positive rate of CK20mRNA, lymph node positive rate of CEAmRNA, Preoperative combined obstruction or perforation, diabetes, tumor invasion depth, distance from the tumor lower margin,the number of lymph nodes under examination, lymph node metastasis after operation with or without chemotherapy (p<0.05). Lymph node positive rate of CK20mRNA, lymph node positive rate of CEAmRNA, preoperative combined obstruction or perforation, tumor invasion depth, distance from the tumor lower margin were independent prognostic factors in colorectal cancer. The positive rate of CK20 in peripheral blood, diabetes, with or without chemotherapy after surgery and the number of lymph nodes under examination were unable to enter the Cox multivariate model.ConclusionThe application of realtime-PCR to detect the expression of CK20mRNA in mesenteric lymph nodes in diagnosis and prognosis system of colorectal cancer is of great significance and value. |
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