| Objective: Colorectal carcinoma is one of the most common malignant tumors in our country. The tendency 0f incidence is increasing. Although certain curative effect is obtained by operation, radiation therapy and chemotherapy, data display that approximately 30% patients died of recurrence and distant metastasis within 5 years. So the study of mechanism of invasion and metastasis about colorectal carcinoma became a focus at present. Many studies displayed that invasion and metastasis of carcinoma is a multiple stepwise and complex process. Degradation of basement membrane around carcinoma is a basic step in the process that carcinoma cells invade fibre and connective tissue metastasis towards distant organism. The loss of basement membrane integrity is closely related to invasion, distant metastasis and poor prognosis. Matrix metalloproteinases (MMPs) secreted by carcinoma tissue may degrade extracellular matrix and basement membrane and induce carcinoma invasion and metastasis. On the other hand, tissue inhibitor of matrix metalloproteinase can suppress its activity and hold back the invasion and metastasis of carcinoma.There are little researches about biological function of MMPs and TIMPs in colorectal carcinoma in country at present. Therefore, we researched expression of MMP-2, TIMP-2, MT1-MMP at gene level in colorectal carcinoma and relationship between them and clinical pathology in order to explore their function in the mechanism of invasion and metastasis. We hope that the data can present the theory reference for MMP inhibitor in colorectal treatment.Methods: 30 cases of colorectal carcinoma specimens and paired normal mucosa were collected from patients operated in the Surgery Department of the Fourth Hospital of HEBEI Medical University during August 2001 to December 2001.The distance from carcinoma to normal mucosa is 10 centimeter. All specimens were collected within half an hour after the operation, fast frozen by the liquid nitrogen and reserve in -80℃ refrigerator. All specimens were verified by pathology. We used the method of acid guanidinium phenol chloroform to extract tissue common RNA, carried out RT-PCR to detect MMP-2, TIMP-2, MT1-MMP and amplified β-actin as internal reference. Products were taken photos and analyze by gelatum imaging system after electrophoresis. We attained product of area and brightness as integral optic density(IOD), regarded the IOD ratio of MMP-2, TIMP-2, MT1-MMP andβ-actin as their expression level. We analyzed and statisticed data by chi-square test, t test, one-way ANOVA, person correlationand multiple stepwise regression analysis.Results: 1. 30 cases (100%) in colorectal carcinoma and 22cases (73.3%) in normal mucosa express MMP-2 mRNA. There was significant difference between them (P<0.01). The expression level of MMP-2mRNA in carcinoma and normal mucosa was 0.3472±0.0429,0.1850±0.0333. There was significant difference between them (P<0.01). There was significant difference (P<0.05) between lymph node metastasis and without lymph node metastasis. The expression level of MMP-2mRNA stepped up in Dukes B, C, and D. There was significant difference (P<0.05) in Dukes D vs. Dukes A, B, and C. There was positive correlation (P<0.05) between expression level of MT1-MMPmRNA and Dukes stage analyzed by multiple stepwise regression. The expression level of MMP-2mRNA in carcinoma was not related to age, sex, tumor size, differentiation and tumor location (P>0.05).2. 29 cases (96.79%) in colorectal carcinoma and 28 cases (93.39%) in normal mucosa express TIMP-2mRNA. There was no significant difference between them (P>0.05). The expression level of TIMP-2mRNA in carcinoma and normal mucosa was 0.7269±0.0496,0.6325±0.0532. There was no significant difference between them (P>0.05).With the progress of Dukes stage, the expression level of TIMP-2mRNA enhanced. There was significant difference (P<0.05) in Dukes D vs. Dukes A, B, and C. There waspositive correlation (P<0.01) between expression level of TIMP-2 mRNA and Dukes s... |
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