The Expreesion And Significance Of PRM1in Human Colon Adenocarcinomas

Tumor is one of the main reasons for human life and health threat. Colon cancer is one high incidence of malignancy in China, accounting for the third in total malignant tumors of the gastrointestinal. Patients with colon cancer increased year by year, and are getting younger and younger. Early detection and early treatment is an important principle in the treatment of cancer.Diagnosis of colon cancer mainly depends on the means of colonoscopy and abdominal CT, so most patients lost opportunity of surgical treatment when they get clearly diagnosis, so look for specific early diagnostic markers and molecular therapeutic targets is very important. Protamine (PRM) is considered to be related with male infertility, the latest research shows that, PRM1may be a new colon cancer/testis (Cancer/Testis, CT) antigen gene. The purpose of this study is to elaborated the significance of PRM1in the clinical diagnosis of colon cancer and the possibility of PRM1as gene therapy targets of colon cancer through the detection of PRM1-mRNA and protein levels change in colon cancer tissue. Specific studies are as follows:1.Experimental Routes:1.1Collecte peripheral blood of patients with colon cancer, the serum was separated; collected colon cancer tissue and normal colon tissues.1.2Compare the differences of PRM1mRNA expression level of colon cancer tissue and normal colon tissue via Real-time quantitative PCR.1.3Agarose gel electrophoresis comparison of normal colon tissue and intestinal tissue content between PRM1-mRNA1.4Analysis sensitivity of PRM1mRNA detection for the diagnosis of colon cancer.1.5Immunohistochemical staining observe PRM1expression in colon cancer tissue1.5.1Compare PRM1expression level colon cancer tissue the and normal colon tissue PRM1expression levels.1.5.2Compare difference of PRM1expression in high differentiated adenocarcinoma, medium differentiated colon adenocarcinoma and poorly differentiated colon adenocarcinoma.1.6Detect serum PRM1levels of patients with colon adenocarcinoma.1.6.1Compare the differences serum levels between patients with colon adenocarcinoma and health controls.1.6.2Compare the differences serum levels between patients who with adenocarcinoma in different stages.1.6.3Compare the differences serum levels between high, medium and poorly differentiated colon adenocarcinoma.1.6.4The positive rate of serum PRM1for colon cancer diagnosis.2.Result2.1The PRM1mRNA expression levels in the colon cancer is higher than that in normal colon tissues The average PRM1-mRNA content of53cases of colon cancer tissue was significantly higher than that in normal colon tissue, P=0.01116. The same results consistent with the results of the quantitative PCR was got through the luminance scanning when quantitative PCR reaction product was subjected to agarose gel electrophoresis and compared PRM1mRNA expression difference between colon carcinoma and normal colon tissue.2.2Sensitivity analysis of PRM1mRNA detection for colon cancer diagnosis PRMl-mRNA expression was defined positive when PRM1-mRNA content in colon cancer tissue was higher or equal to2times than that in normal colon tissue. Statistics results showed that the PRM1mRNA positive rate was96.23%(52/53)2.3Immunohistochemical staining assay detected PRM1expressions in colon cancer tissuePRM1expression and tissue distribution characteristics was detected via immunohistochemistry. The results showed that PRM1positive staining particles was found in the cytoplasm of colon cancer cells and glandular cavity surrounded by cancer cells. PRM1staining intensity in poorly differentiated adenocarcinoma cells was significantly higher than the well-differentiated adenocarcinoma cells.2.4ELISA test detected serum PRM1level of patients with colon cancer2.4.1Compare serum PRM1level with colon cancer with that in healthy people.Detected serum PRM1levels in patient with colone cancer and healthy adults by ELISA assay, the results showed that the serum PRM1level of patients with colon cancer were significantly higher than that in healthy adults, P<0.05.2.4.2The positive rate of serum PRM1used for colon cancer diagnosisUsing the confidence interval for the group of healthy subjects as the cut-off value, the sensitivity of PRM1as markers for colon adenocarcinoma were calculated to investigate its diagnostic efficacies.Ap-value≤0.05was considered to be statistically significant.The results showed that the serum positive rate of patients with colon cancer was80%(24/30)2.4.3Compared PRM1levels of patients with different stages colon cancerCompare serum PRM1level of patients with T1NXMX、T2NXMX、T3NXMX colon cancer, the results showed that there was no significant differences in serum PRM1level of patients who in different colon stages.2.4.4Compared PRM1levels of patients with different differentiation Compared serum PRM1level according to Pathological differentiation of colon adenocarcinoma cells. The results showed the PRM1level of patients with poorly differentiated colon cancer was higher than well-differentiated colon cancer, P<0.05, there was no differents between well-differentiated colon cancer and moderate-differentiated colon cancer, moderate-differentiated colon cancer and poorly differentiated colon cancer.3. Conclusions3.1Real-time PCR showed that colon adenocarcinoma PRM1mRNA levels were significantly higher than that in normal colon tissue.3.2Immunohistochemical staining showed PRM1protein expression found in colon cancer cells and surrounded by glandular cavity, PRM1expression and differentiation of colon cancer cells related to:poorly differentiated adenocarcinoma PRM1expression levels higher than moderately differentiated adenocarcinoma, well differentiated adenocarcinoma and normal colon tissue PRM1no expression. Tip PRMl possible role in tumor cells through paracrine manner.3.3PRM1peripheral serum levels of colon cancer patients than in healthy control group, suggesting PRM1may be secreted into the blood by the action of the endocrine organs in other ways.3.495%confidence interval in healthy control group to determine the level of serum PRM1levels in peripheral blood PRM1positive rate of80%for the diagnosis of colon cancer.