A Primary Study Of The Expression Of E-CD,TIMP-3,MMPS, And The Mechanism Of Methylation In Colon Cancer Carcinogenesis And Metastasis

Purpose: In order to discover the molecular changes and mechanism in colon cancer development, invasion and metastasis process and provide the academic base and treatment target for colon cancer ,we examined E-CD , TIMP-3 ,MMP-2, MMP-9 expression and 5' CpG inslands' methylation status of CDH1 and TIMP-3 gene promoter region in no-tumor colon disease , adenmoma and colon cancer samples, and observed the relationship of the expression of protein and the methylaiton status with the biological behavior of cordon cancer, and the relationship of the methylation status with the expression of protein. Materials and Methods: 16 no tumor disease in colon,32 adenoma,35 colon cancer specimens were collected at random. The E-CD, TIMP-3, MMP-2, MMP-9 were detected by SP immunohistochemistry. The 5' CpG islands' methylation status of CDH1 and TIMP-3 gene were detected by methylation-specific PCR (MSP). The relationship of the methylation status with the expression of relevant protein was analysed. The relationship of the methylation status and the expression of protein with the biological behavior of conlon cancer was analysed. Results:1.the expression of E-CD and the methylation status of CDH1 and their relationship with biological behavior of colon cancer 1.1 the expression of E-CD and its relaitonship with biological behavior of coloncancer1.1.1 the expression of E-CD In 16 no tumor colon disease, 32 adenoma and 35 colon cancer, the expression ofE-CD respectively is 100%(16/16), 96.88%(31/32) and 65.71 %(23/35). Theexpression of E-CD in colon was reduced than which in no tumor colon diseaseand adenoma (PO.01).1.1.2 the relationship of the expression of E-CD with the biological behavior ofcolon cancerThe expression of E-CD was different in different histology types of colon cancer.The expression of E-CD in the group of mutinous adenocarcinoma,signet ring cellcarcionma and poor differentiated carcinoma was reduced than in the group oftubular adenocarcinoma and papillary adenocarcinoma^ <0.01).The expression of E-CD in the group of poor differentiated was reduced than inthe group of high and middle differentiated (P <0.01).The expression of E-CD in the group of Duck's C+D stages were reduced than inthe group of Duck's A+B stages (P <0.01).The expression of E-CD in the group of positive lymph metastasis was reducedthan in the group of negative lymph meatstasis.1.2 the 5'CpG islands methylation status of CDH1 gene promoter region and itsrelationship with biological behavior of colon cancer 1.2.1 the 5'CpG islands methylation status in CDH1 gene promoter region The5'CpG CpG islands methylation status in CDH1 gene promoter region of 16 no tumor colon disease and 32 adenoma all were 0.00%, and 25.71%(9/35) in colon cancer. The frequency of CpG islands methylation of CDH1 gene promoter region in colon cancer is higher than in no tumor colon disease and in adenoma (P1.2.2 the relationship of the 5'CpG islands the methylation status in CDH1 gene promoter region with the biological behavior of colon cancer There were no relationship of the 5'CpG islands methylation status in CDH1 gene promoter region with types of histology and differentiation in colon cancer (PX).O5).The 5'CpG islands methylation status in CDH1 gene promoter region in Duck'sA+B, positive lymph node metastasis were reduced than in Duck's C+D, negativelymph node metastasis (P <0.01).1.3 the 5'CpG islands methylation status in CDH1 gene promoter region ispositive correlation with E-CD protein expression in colon cancer (P <0.05).2. the expression of TTMP-3, the methylation status of TIMP-3 and theirrelationship with biological behavior of colon cancer2.1 the expression of TIMP-3 and its relaitonship with biological behavior ofcolon cancer2.1.1 the expression of TIMP-3hi 16 no-tumor colon disease, 32 adenoma and 35 colon cancer the expression of TIMP-3 respectively is 12.50% (2/16) ,28.13% (9/32), and 82.86% (29/35) . The expression of TIMP-3 in colon cancer was higher than which in no tumor colon disease and adenoma (P <0.01).2.1.2 the relationship of the expression of TIMP-3 with the biological behavior of colon cancerThere in no relationship of the expression of TIMP-3 with differentiation (P <0.05). The expression of TIMP-3 was different in different histology types ofcolon cancer. The expression of TIMP-3 in the group of mutinousadenocarcinoma,signet ring cell carcionma and poor differentiated carcinoma wasreduced than in the group of tubular adenocarcinoma and papillaryadenocarcinoma {P < 0.05).The expression of TIMP-3 in the group of Duck's C+D stages was reduced than inthe group of Duck's A+B stages (P < 0.05).The expression of TIMP-3 in the group of positive lymph metastasis was reducedthan the group of negative lymph meatstasis {P <0.01).2.2 the 5'CpG islands methylation status in TIMP-3 gene promoter region and its relationship with biological behavior of colon cancer2.2.1 the 5'CpG islands methylation status in TIMP-3 gene promoter region The 5'CpG island methylation status in TIMP-3 gene promoter region of 16 no tumor colon disease and 32 adenoma all were 0.00%, and 20.00% (7/35) in colon cancer. The 5'CpG islands methylation status in TMP-3 gene promoter region in colon cancer is higher than in no tumor colon disease and in adenoma (P <0.01).2.2.2 the relationship of 5'CpG islands methylation status in TIMP-3 gene promoter region with the biological behavior of colon cancerThere were no relationship of the 5'CpG island methylation status in TIMP-3differentiation in colon cancer (P > 0.05).The 5'CpG islands methylation status in TIMP-3 gene promoter region in positivelymph node metastasis were higher than in negative lymph node metastasis (P <0.05).The 5'CpG islands methylation status in TIMP-3 gene promoter region in Duck'sC+D stages were higher than in Duck's A+B stages (P <0.05).The 5'CpG island methylation status in TIMP-3 gene promoter region in thegroup of mucinous adenocarcinoma, signet ring cell carcionma and poordifferentiated carcinoma is higher than in the group of tubular adenocarcinomaand papillary adenocarcinoma (P < 0.05).2.3 The 5'CpG islands methylation status in TIMP-3 gene promoter region is positive correlation with TDVIP-3 protein expression in colon cancer (P < 0.01).3. the expression of MMP-2,MMP-9 and their relationship with biologicalbehavior of colon cancer3.1 the expression of MMP-2,MMP-9In 16 no tumor colon disease the expression of MMP-2 and MM-9 respectively is12.50%(2/16), 6.25% (1/16) .In 32 adenoma, the expression of MMP-2 andMMP-9 respectively is 21.88% (7/32) ,15.63% (5/32) .In 35 colon cancer the expression of MMP-2 and MMP-9 respectively is 74.29% (26/35) ,77.14% (27/35 ). The expression of MMP-2 and MMP-9 in colon cancer was higher than which in no tumor colon disease and adenoma (P <0.01).3.2 the relationship of the expression of MMP-2, MMP-9 with biological behavior of colon cancerThere were no relationship the expression of MMP-2 ^ MMP-9 with differentiation and histology typesCP >0.05).The expression of MMP-2,MMP-9 in the group of mucinous adenocarcinomas, signet ring cell carcionmas and poor differentiated carcinomas is higher than in the group of tubular adenocarcinomas and papillary adenocarcinomas (P < 0.01). The expression of MMP-2 and MMP-9 in Duck's A+B stages was reduced than Duck's C+D stages (P <0.05).The expression of MMP-2 and MMP-9 in positive lymph node metastasis was higher than in negative lymph node metastasis (P <0.05).4.The relationship of the expression of E-CD with TEVIP-3 and TIMP-3 with MMP-2,MMP-9 in colon cancerThe expression of E-CD and TIMP-3 is positive correlation (P<0.01). The expression of TIMP-3 was negative correlation with the expression of MMP-2 and MMP-9 (PO.01). Conclusions:1 .The expression of E-CD in colon cancer was reduced than in no tumor disease and adenoma. So E-CD plays an important role in carcinogenesis of colon cancer. 2.No expression of E-CD in colon cancer was correlation with histology types, poor differentiation, Duck's C+D stages, lymph node metastasis of colon cancer. It show that E-CD play a important role in differentiation, invasion and metastasis of colon cancer. Expression of E-CD is a important parameter for judgement of...