The Expression And Significance Of PRM1in Human Colon Adenocarcinoma

Colon cancer is one high incidence of malignancy in China, accounting for thethird in total malignant tumors of the gastrointestinal. Patients with colon cancerincreased year by year, and are getting younger and younger. Diagnosis of coloncancer mainly depends on the means of colonoscopy and abdominal CT, so mostpatients lost opportunity of surgical treatment when they get clearly diagnosis, solook for specific early diagnostic markers and molecular therapeutic targets is veryimportant. Protamine （PRM） is considered to be related with male infertility, thelatest 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 ofPRM1in the clinical diagnosis of colon cancer and the possibility of PRM1as genetherapy targets of colon cancer through the detection of PRM1-mRNA and proteinlevels change in colon cancer tissue. Specific studies are as follows:Experimental Routes:1.1Collecte peripheral blood of patients with colon cancer, the serum wasseparated; collected colon cancer tissue and normal colon tissues.1.2Compare the differences of PRM1mRNA expression level of colon cancertissue and normal colon tissue via Real-time quantitative PCR.1.3Analysis sensitivity of PRM1mRNA detection for the diagnosis of coloncancer.1.4Immunohistochemical staining observe PRM1expression in colon cancertissue1.4.1Compare PRM1expression level in colon cancer tissue and normalcolon tissue PRM1expression levels.1.4.2Compare difference of PRM1expression in high differentiated adenocarcinoma, medium differentiated colon adenocarcinoma andpoorly differentiated colon adenocarcinoma.1.5Compare the differences of PRM1expression level between colonadenocarcinoma and normal colon tissue by western blot.1.6Detect serum PRM1levels of patients with colon adenocarcinoma.1.6.1Compare the differences serum levels between patients with colonadenocarcinoma and health controls.1.6.2Compare the differences serum levels between patients who withadenocarcinoma in different stages.1.6.3Compare the differences serum levels between well, moderate andpoorly differentiated colon adenocarcinoma.1.6.4The positive rate of serum PRM1for colon cancer diagnosis.1.7ELISA assay detected serum CEA level in colon cancer.1.8Compare the sensitivity of serum PRM1, CEA for diagnosis colonadenocarcinoma.2. Result2.1The PRM1mRNA levels in the colon cancer is higher than that in normalcolon tissuesThe average PRM1-mRNA content of53cases of colon cancer tissue wassignificantly higher than that in normal colon tissue, P=0.01116. The same resultsconsistent with the results of the quantitative PCR was got through the luminancescanning when quantitative PCR reaction product was subjected to agarose gelelectrophoresis and compared PRM1mRNA expression difference between coloncarcinoma and normal colon tissue.2.2Sensitivity analysis of PRM1mRNA detection for colon cancer diagnosisPRM1-mRNA expression was defined positive when PRM1-mRNAcontent in colon cancer tissue was higher or equal to2times than that in normalcolon tissue. Statistics results showed that the PRM1mRNA positive rate was96.2% （51/53）2.3Immunohistochemical staining assay detected PRM1expressions incolon cancer tissuePRM1expression and tissue distribution characteristics were detected viaimmunohistochemistry. The results showed that PRM1positive staining particleswere found in the cytoplasm of colon cancer cells and glandular cavity surroundedby cancer cells. PRM1staining intensity in poorly differentiated adenocarcinomacells was significantly higher than the well-differentiated adenocarcinoma cells.2.4Detected PRM1expression in colon cancer tissue via western blotThe experiments used western blot to detect PRM1expression levels of coloncancer tissue and normal colon tissue. The results showed that the PRM1level incolon cancer tissue were far higher than that in the normal colon tissue. Western blotexperiment together with the immunohistochemistry experiment proved the PRM1expression level was higher in colon cancer tissue than that in normal colon tissue.2.5ELISA test detected serum PRM1level of patients with colon cancer2.5.1Compare serum PRM1level with colon cancer with that in healthypeople.Detected serum PRM1levels in patient with colone cancer and healthy adultsby ELISA assay, the results showed that the serum PRM1level of patients withcolon cancer were significantly higher than that in healthy adults, P <0.05.2.5.2The positive rate of serum PRM1used for colon cancer diagnosisUsing the confidence interval for the group of healthy subjects as the cut-offvalue, the sensitivity of PRM1as markers for colon adenocarcinoma were calculatedto investigate its diagnostic efficacies.Ap-value≤0.05was considered to bestatistically significant.The results showed that the serum positive rate of patientswith colon cancer was80%（24/30）2.5.3Compared PRM1levels of patients with different stages colon cancerCompare serum PRM1level of patients with T₁N_XM_X、T₂N_XM_X、T₃N_XM_X colon cancer, the results showed that there was no significant differences in serumPRM1level of patients who in different colon stages.2.5.4Compared PRM1levels of patients with different differentiationCompared serum PRM1level according to Pathological differentiation of colonadenocarcinoma cells. The results showed that there was no significant differencesin serum PRM1level of patients with different differentiation, P>0.05.2.6Detected serum CEA level of patients with colon cancer via ELISA testUsing ELISA test to detect serum CEA level of patients with colon cancer andhealthy adult, the results showed that the average serum CEA level of patients withcolon cancer was significantly higher than that of healthy adults （P<0.05）2.7serum PRM1, CEA detection for colon cancer diagnostic sensitivitySerum CEA was commonly as a serological diagnostic marker for colon cancerand other tumors, so we compared the diagnostic sensitivity of serum PRM1withCEA as a colon cancer diagnostic marker, the results showed that the diagnosispositive rate of PRM1was80%（24/30） and positive rate of CEA was30%（9/30）.3. Conclusions3.1PRM1mRNA levels in colon cancer were significantly higher than that innormal colon tissue.3.2PRM1positive staining particles were found in the cytoplasm of coloncancer cells and glandular cavity surrounded by cancer cells. PRM1performs bothautocrine and paracrine function.3.3PRM1level in colon cancer tissue was far higher than that in the normalcolon tissue with western blot.3.4The serum PRM1level of patients with colon cancer is significantly higherthan that of healthy adults. PRM1performs endocrine function.3.5Serum PRM1level is more sensitive than serum CEA.