Significance Of The Expressions Of RNA-binding Protein And MRNA In Colorectal Cancers

Objective: In this study, we detect the expression levels of RBM3protein and mRNA in normal colorectal mucosal tissues and cancer tissues,by Immunohistochemistry and real-time PCR, and to analyze the relationshipbetween clinic pathological parameters, and to explore the possiblemechanism of RBM3in the occurrence, development, invasion and metastasisof colorectal cancers to provide more powerful experimental data andtheoretical support for clinical diagnosis,treatment, and prognostic evaluation.Methods:1. To detect the RBM3expression in normal colorectal mucosal tissues and cancer tissues by Immunohistochemistry methods.1.1Material collected:Collect252cases of tissue samples embedded inparaffin blocks(Pathology), which were surgical removal from colorectalcancer patients in Tangshan Gongren Hospital, from May2010to November2011, and228cases were selected by screening. The cases which collected,patients were not receiving chemotherapy and radiotherapy treatmentpreoperative. Study subjects were divided into two groups: normal mucosatissue group and cancer tissue group.228cases of normal mucosa group and228cases of cancer tissue group. Patients with a total of228cases, including143cases of male and85cases of female, aged27to89years, with anaverage age of61.94±11.12years. Including94cases of colon cancer,134cases of colorectal cancer, and they are6cases of Dukes’A,145cases ofDukes’B,75cases of Dukes’C,2cases of Dukes’D, and39cases of TNM Ⅰ,116cases of TNM Ⅱ,70cases of TNM III,2cases of TNM Ⅳ,151caseswithout lymph node metastasis and lymph node metastasis in77cases,42cases of mucinous adenocarcinoma and186cases of non-mucinousadenocarcinoma. The pathological diagnosis of all cases have already been read and referral by more than two deputy director physicians, includinghistological type, grade, and lymph node metastasis. The clinical andpathological data from medical records room and Pathology.During theImmunohistochemistry, normal mucosa shedding84cases and cancer tissuesshedding6, because of tissue repairing and sliced off, and left canceroustissue222cases,normal mucosa tissue144cases.1.2Production of Organization chip: Collect cases of patients with HEstaining and postoperative tissue samples embedded in paraffin blocks, todetermine the area of HE staining of normal mucosa and cancer tissue (Markpen notes) under microscopes, and mark the corresponding area of normalmucosal tissue and cancerous tissue samples embedded in paraffin blocks,punch in the blank wax block with12#bone marrow puncture needle and takethe tissue with16#bone marrow aspiration needle, making tissue chip.1.3Detect RBM3protein expression in normal colorectal mucosa andcarcinoma by immunohistochemical method, and analyze the relationshipbetween the RBM3protein and colorectal clinicopathological parameters.2Detect the expression of RBM3mRNA in normal colorectal mucosaand cancer tissues by Real-time PCR.2.1Collect fresh samples of colorectal cancer30cases in2011, TangshanGongren Hospital, including13males and7females, aged from40to88years’old, with an average age of61.95±13.77years.10cases of coloncancer, rectal cancer10cases,3cases of the Dukes’A,7cases of Dukes’B,8cases of Dukes’C,2cases of Dukes’D. Randomly selected10cases ofnormal tissues adjacent to cancer as a normal control,5males,5females, aged41to74years, with an average age of53.40±9.05years. In these30samples,20cases of cancer tissue,10cases of normal mucosa tissue (Notumor infiltration confirmed by pathology). Assay RBM3mRNA expressionin normal mucosa and colorectal cancer tissue by real-time PCR.3Statistical methods:SPSS14.0software was used for experiment dataresults’ statistical analysis, X2test was used for qualitative data, quantitativedata were analyzed by T-test or F analysis, using spearman correlation studies, P <0.05was considered statistically significant.Results:1. The expression of RBM3protein and mRNA in normal colorectalmucosa and cancer tissue.1.1Positive rate of RBM3protein expression in normal mucosa groupwas47.2%(68/144), and was82.9%(184/222) in the cancer group. Thestatistical analysis of the difference between the two groups was statisticallysignificant (P <0.05). That the RBM3protein expression in cancer tissues wassignificantly higher than the normal mucosa group.1.2The relative ratio of the RBM3mRNA in normal mucosa group was0.0025±0.0013, and was0.0044±0.0008in cancer tissue, the difference of thetwo groups was statistically significant (P <0.05) by statistical analysis. TheRBM3mRNA expression in the cancer tissue group was significantly higherthan its expression in normal mucosal tissue group.2The relationship between the expression of RBM3protein and mRNAand their clinical and pathological features in colorectal cancer tissue.2.1The positive rate of RBM3protein expression in colorectal cancer inDukes A+B was78.9%(116/147), and was90.7%(68/75) in Dukes C+Dperiod. The statistical analysis of the difference between the two groups wasstatistically significant (P <0.05). That is, the positive rate, RBM3proteinexpression in Dukes’C+D in colorectal cancer, was significantly higher thanthat of Dukes’A+B. And about RBM3protein positive expression incolorectal cancer, there was no statistically significant (P>0.05) withdifferent age, gender, lymph node metastasis, tumor size, histological type,depth of invasion, or degree of differentiation.2.2RBM3mRNA positive expression in colorectal cancer tissue therewas no statistically significant (P>0.05) with different gender, age, lymphnode metastasis, histological type, or degree of differentiation.Conclusion:1. RBM3protein expression in colorectal cancer tissue was significantlyhigher than that in the normal mucosa, RBM3related with the early colorectal; 2. RNM3protein’s high expression may be relevant to the progress ofcolorectal cancer;3. The detection of RBM3protein in colorectal cancer tissues contributeto the development of colorectal cancer, and helpful to judge its biologicalbehavior.