Verification Of Colorectal Cancer And Liver Metastasis Related Genes Using RT-PCR Method

Objective:To verify genes screened by gene expression profiling chips which are possibly related to colorectal cancer and liver metastasis of colorectal cancer(LMCC) by means of real-time PCR.Methods:Between Mar 2004 and Oct 2007,80 patients,who underwent colorectal cancer surgery in colorectal professional group of Zhongshan Hospital, were assigned to research groups.Four groups of colorectal normal tissue (n=20),colorectal cancer without liver metastasis(n=20),colorectal cancer with synchronia liver metastasis(n=20),colorectal cancer with heterochronia liver metastasis(n=20) were built up.Correspondence cycle threshold(CT) value of 84 genes expressed in the 80 specimen were analyzed using real-time PCR method based on SYBR green.We compared the expressed quantity between colorectal cancer without liver metastasis and synchronia liver metastasis,colorectal cancer without liver metastasis and heterochronia liver metastasis,synchronia liver metastasis and heterochronia liver metastasis,colorectal normal tissue and colorectal tumor tissue.T test was used to compare the difference among groups, 2^{-△△CT} method was used to analyze the correspondence expressed quantity.Results:Four differentially expressed genes were found probably related to colorectal cancer liver metastasis,including three up regulation genes and one down regulation genes.SRC gene is up regulated 2.2 times in colorectal cancer with synchronia liver metastasis(p=0.016),AZGP1 is 2.4 times(p=0.025) up regulated in synchronia liver metastasis,IQCA is 3.5 times(p=0.015) up regulated in synchronia liver metastasis.HCA112 gene is down regulated 0.61 times(p=0.04) in colorectal cancer with heterochronia liver metastasis. 24 differentially expressed genes were found probably related to colorectal cancer.20 genes are down regulated such as EGFL6,HRASLS,HFE, DTX1,SLC16A7,MAF,ST70T1,PLCL2,ISL1,SLC30A7,MAGED4,CAMK2N1,TDGF1, IL28RA,CYP51A1,SCIN,CDKL1,AQP11,GPR160 and CUGBP1,4 genes are up rugulated such as PLEKHH1,SATB2,AP1G2 and MFSD2.Conclusion"1) Up regulation of gene SRC,AZGP1 and IQCA in colorectal cancer probably have relationship with colorectal cancer synchronia liver metastasis.2) Down regulation of gene HCA112 probably have relationship with heterochronia liver metastasis.3) Down regulation of gene EGFL6,HRASLS,HFE,DTX1,SLC16AT,MAF, STT0T1,PLCL2,ISL1,SLC3OAT,MAGED4,CAMK2N1,TDGF1,IL28RA,CYP51A1, SCIN,CDKL1,AQP11,GPR160 and CUGBP1 may concerned with the development of colorectal cancer.4) Up regulation of gene PLEKHH1,SATB2,AP1G2 and MFSD2 may concerned with the development of colorectal cancer.5) All genes need further research.