Screening And Identification Of Differentially Expressed Genes In Patients With Pre - Diabetes

Part one Application of human whole genome expression profile chip in screening different gene of pre-diabetes Purpose: Diabetes has become a chronic disease that is a serious threat to huamn health.With the increaseing of pre-diabetes patients,diabetes population is growing too.And the complications of diabetes is also increasing.It will seriously reduce the patient’s life quality and increase economic burden of social and family.Pre-diabetes is a middle stage between normal glucose tolerance and diabetes,and a early stage in the disorder of glucose metabolism. Most pre-diabetes patients don’t show typical symptoms of diabetes,such as drinking much water,eating much food,polyuria and weight loss.Even some show no symptoms. So it can not cause people’s alarm and attention.Many people do not consider them as high risk groups.Pre-diabetes is a reversible process. If people can find gene changes in the early stage and give targeted intervention,we can decrease the risk of diabetes.In recent years,people have already realized that pre-diabetes is a complex process with multiple genes involed.The early detection of pre-diabetes genes can help us find the changes which may have been ignored or cann’t be found by histological method.These genetic changes may indicate diabetes.This study will screen differential gene expression between prediabeticpatients and normal person.We will analysis the role of differentially expressed genes in pathogenesis of pre-diabetes and lay a the new theory foundation for the further study of pre-diabetes gene diagnosis and gene targeted intervention. Methods: Peripheral blood samples were collected from 17 cases of pre-diabetes and 13 cases of healthy human.Use Agilent human genome-wide expression profile chip with 4*44K technology to analysis the expression of samples’ m RNA, use the t test and difference between multiple to choose the differentially expressed genes,then get the genes that associate mostly with the onset of diabetes and that have the most obvious different expression by gene function screening from a large number of the differentially expressed genes. Results: For control group, 13252 different genes are selected when P<0.05,,including 7414 up-regulated genes and 5838 down-regulated genes,65 probes with the fold changes above 2 times are selected,including 37 up-regulated and 28 down-regulated,and 474 probes with the fold change above 1.5 times are selected,including 279 up-regulated and 195 down-regulated.Differentially expressed genes are mainly involved in cell cycle,immune response,metabolism and other biological processes,related to cell cycle pathway,niacin and nacinamidemetabolism,nitrogen metabolism,purine metabolism,pyrimidine metabolism,histidine metabolism,MAPK signal transduction pathway, melanoma and other signal transduction pathway. Conclusions: We can effective screen the differentially expressed genes which related to pre-diabetes by applying human genome-wide expression profile chip.Part two Identification the differentially expressed genes of Pre-diabetes Purpose: To remove errors and false positive of the genen chipi experiment results, chooseing genes of expression apparent and related to glucose metabolism by real-time PCR analysis..Observe it’s expression in m RNA between pre-diabetes group and the normal group. Via preliminarying screening and verificating the differentially expressed genes which relate to pre-diabetes, can early discovery the potential risk of pre-diabetes.To provide evidence for early diagnosis and targeted intervention of pre-diabetes.Methods: Using Real-time fluorescent quantitative PCR technique to detect the expression intensity in m RNA between pre-diabetes and healthy human, take the endogenous genes as internal. Results: The results of Real-time PCR showing, the expression quantity of RNF182 and LTF of pre-diabetes are higher than the normal group(P<0.05), with the trend of the chip results.In addition,the expression of ENPP1 and TCF7L2 in pre-diabetes group is higher than normal group. Conclusions: The results after gene chip verification is reliable, but still can’t exclude the false positive results in gene chip. We can select the meaningful differentially expressed genes by using gene chip, but the results of it still needs further verification.ENPP1 and TCF7L2 may be disease genes of pre-diabetes.But it still need to be further validation of large sample.