The Research Of Normal Intesinal Mucosa MRNA Express Spectrum On The Patients With Colon Cancer

Background and Purpose:Nowadays, there are plenty of researches in which suggest that the developing process of tumor and the abnormal condition are extremely related. By observing the Tumor mRNA's expression profile and focusing on some special cases of mRNA's expression, we found that this is positively influenced diagnose of tumor in clinical territory. The mortality rate that casued by Colorectal Cancer has been taken the second place of the total mortality rate of female, and the fourth place of male in our country. However, there are no Gene level can be consider as a symbol for doctors to diagnose this canceer. It has been recognised by a large amount of studies that the occurrence of Colorectal Cancer has closed relationship with immunity of human-being. The aim of this research is to anaylising the Colorectal Cancer and the changing of mRNA's expression. By discussing the mRNA's Chip express spectrum technique ,and by testing the mRNA expression ; to classify the mRNA in which is related to immunity. And studying those mRNA for finding out the relationships between the expression and the occurrence of Colorectal Cancer; so that we are able to analysing the early stage of diagnosing Colorectal Cancer.Patients and Methods:18 patients with colon cancer that have been confirmed by the pathological. Get normal mucosa samples from 43 parts of anal(that is 5cm far from the tumor part), colon and colorectal(and they are named by"a b c"in the study),and then put the samples into mRNA protective agent and stored them in the fridge with the temperature of—80℃.Using RNeasy mini kit to get mRNA from the samples we get before.In order to get qualified mRNA and study them,we use the all wavelengths ultrovilot/visible scan spectrophotometer and 6000 Nano Kit to test the concentration and qualification of the mRNA. Dyeing the mRNA( that are qualified) with cy3, and take the qualified dyeing mRNA into our study group. Finally,with the method of Whole Human Genome (4x44k)Oligo Microarray Kit to hybidize with mRNA,and put the hybidized chips into machine of Agilent Microarray Scanner.what's more,they will also receive the test of Quality Ccontrol system,after so many tests, the pictures that scanned from the qualified chips will be analysed by the system of GeneSpring GX 11.0.2.Results:Using RNeasy mini kit to abstract mRNA,and with NanoDrop ND-1000 to check them, we find that there are 3 patients,9 samples in all, whose mRNAs are totally more then 50ng.And afer the test of Agilent RNA 6000 Nano Kit we find that all the fragments mRNA's integrity scores are upper 5(which means that the integrity of the abstracted mRNA is not destroyed,and the high purity can be used for the test of gene chip.)Dyeing the samples with Cy,and use NanoDrop ND-1000 to test the dyeing rate, after caculation ,it shows that the dyeing is qualified,and the samples are suited to the request of gene chip test.Hybridizing Whole Human Genome (4x44k)Oligo Microarray Kit with sample mRNAs,we totally find 41093 mRNAs,and after the test of Quality Ccontrol system, they are all qualified.Using Oneway-ANOVA method,we find 2472 function related mRNAs,p≤0.05.And with GO Analysis system we find 300 Extracellula function related mRNA, p≤0.1.After anlysing the data from GeneSpring GX 11.0.2 system we can find that, within the 41 immune related mRNAs,there are 21 mRNAs whose total represenation rate is positive correlation with the distance of tumor, and 5 mRNA's total representation rate is negtive correlation with the distance of tumor,and 15 mRNAs total representation rate is not obvious.Conclusions:The results shows that in the mucosa of the colon and the rectal of the patients with colon cancer ,there are some immune related mRNA,and they show different expression rate in different parts of the colon and the rectal,which testify that there are some closely connection between the cause of tumor and body immunity.what's more,some immune related mRNAs expression rate shows that the far it is from the tumor part,the higher expression rate it is.so, we consider that there is an'escape phenomenon'of some immune related mRNA during the tumor cause process ,and gene testing helps to find the phenomenon.If we find some method to interfere this phenomenon,it may do help in clinic treatment of colon cancer.Also,with the help of mRNA chip technology, we can observe the expression of immune related mRNA,and it may help in early diagnosis of colon cancer.