【作者】 吕世旭；

【导师】 许剑民； 卢大儒； 钟芸诗； 韦烨； 范薇薇； 任黎；

【作者基本信息】 复旦大学 ， 普通外科， 2009， 硕士

【摘要】 目的:运用Real time-PCR方法开展对以往全基因芯片筛查所得的结直肠癌和结直肠癌肝转移相关基因进一步的验证研究。方法:选取复旦大学附属中山医院结直肠癌专业组2004年3月～2007年10月手术的结直肠正常组织及结直肠癌组织冻存标本共80例,分为结直肠正常组织组（n=20）,结直肠癌无肝转移组（无肝转移组,n=20）,结直肠癌同时性肝转移组（同时性肝转移组,n=20）与结直肠癌异时性肝转移组（异时性肝转移组,n=20）。采用基于SYBR green的荧光染料掺入法real-time PCR检测所有标本中84个基因（源自以往全基因芯片筛查所得的结直肠癌和结直肠癌肝转移相关基因）的相对表达量,比较无肝转移组与同时性肝转移组、无肝转移组与异时性肝转移组、同时性肝转移组与异时性肝转移组基因表达差异。另外,分别比较结直肠正常组织组与无肝转移组、同时性肝转移组及异时性肝转移组之间的基因表达差异。运用t检验方法对各组△CT值行组内比较,2^-△△CT法分析基因相对表达量。结果:在结直肠癌肝转移相关基因研究中共得到4个差异基因,其中表达上调基因3个,下调基因1个。同时性肝转移组较无肝转移组SRC基因的表达升高2.2倍（p=0.016）;同时性肝转移组AZGP1基因的表达较无肝转移组升高2.36倍（p=0.025）;同时性肝转移组IQCA基因较无肝转移组上调3.50倍（p=0.015）;HCA112基因在异时性肝转移组较无肝转移组下调0.61倍（p=0.040）。在所有结直肠癌组织与结直肠正常组织基因表达差异研究中,共获得24个差异基因,其中下调基因20个,分别为EGFL6、HRASLS、HFE、DTX1、SLC16A7、MAF、ST70T1、PLCL2、ISL1、SLC30A7、MAGED4、CAMK2N1,TDGF1、IL28RA、CYP51A1、SCIN、CDKL1、AQP11、GPR160、CUGBP1;上调基因4个,分别为PLEKHH1、SATB2、AP1G2、MFSD2。结论1 SRC,AZGP1,IQCA基因在结直肠癌组织中的上调表达可能与同时性肝转移相关;2 HCA112基因在结直肠癌组织中的下调表达可能与异时性肝转移相关;3 EGFL6、HRASLS、HFE、DTX1、SLC16A7、MAF、ST70T1、PLCL2、ISL1、SLC30A7、MAGED4、CAMK2N1、TDGF1、IL28RA、CYP51A1、SCIN、CDKL1、AQP11、GPR160、CUGBP1基因的下调表达可能与结直肠癌的发生有关;4 PLEKHH1、SATB2、AP1G2、MFSD2基因的上调表达可能与结直肠癌的发生有关;5上述基因的功能,与结直肠癌及结直肠癌肝转移的关系需要进一步的验证研究。更多还原

【Abstract】 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.更多还原