| ObjectiveBreast cancer is one of the most common malignant tumors in our country. The tendency Of incidence is increasing. Although certain curative effect is obtained by operation, radiation therapy and chemotherapy, data display that approximately 20% patients died of recurrence and distant metastasis within 5 years. So the study of mechanism of invasion and metastasis about breast cancer became a focus at present. Many studies displayed that invasion and metastasis of cancer is a multiple stepwise and complex process. Degradation of basement membrane around cancer is a basic step in the process that cancer 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 cancer tissue may degrade extracellular matrix and basement membrane and induce cancer invasion and metastasis. On the other hand, tissue inhibitor of matrix metallo-proteinase can suppress its activity and hold back the invasion and metastasis of cancer. There are little researches about biological function of MMPs and TIMPs in breast cancer in country at present. Therefore, we researched expression of MMP -2, TIMP -2, MT1 - MMP at protein level in breast cancer 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 breast treatment.Methods64 cases of breast cancer specimens and 30 cases of breast benign tumor were collected from patients operated in the Oncology Department of the First Hospital affiliated to China Medical University during May 2004 to December 2004. 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. The expression of MMP - 2, TIMP - 2 and MT1 - MMP on 64 cases of breast cancer were detected immunohistochemically, and then 34 cases of breast cancer specimens with the co - expression of MMP - 2, TIMP -2 and MT1 - MMP were analyzed by the Image Analysis Software of Immunohis-tochemistry quantitative test. Furthermore, the expression of MMP - 2, TIMP -2 and MT1 - MMP on 60 cases of breast neoplasm were determined by Western Blot, including 30 breast cancers( study group) and 30 benign breast tumors. Products were taken photos and analyzed by imaging system after electrophore-sis. We attained product of area and brightness as integral optic density (IOD) , the IOD of MMP -2, TIMP -2, MT1 - MMP were regarded as their expression level. The quantitative data from Immunohistochemistry and Western blot were analyzed by the statistical analysis methods including t test and one - way ANO-VA, which were analyzed by SAS6. 12 ( Statistical Analysis System Version6. 12) and the significant size of test of this study was validated as avalue of 0.05.ResultsTo use Immunohistochemistry to detect the expression of MMP - 2, TIMP -2 and MT1 - MMP in the 64 cases, the number of co - positive expression is 34 cases, the data of MMP -2,TIMP -2 and MT1 - MMP analyzed by the Image Analysis System is 3. 11 ±0. 74,2. 86 ±0. 65,3. 06 ± 0. 88 respectively. Positive expression is mainly locate in the cancer cell intracytoplasm, to show diffusive distribution of brown granulo, negative expression is in the cancer cell nucleus , less expression is in the interstitial cell. And then the expression of MMP— 2, TIMP - 2 and MTl - MMP in breast cancer was divided into groups and layers by the clinical patho -factors of breast cancer( T stage, N stage, TNM stage, ER, PR and C - erbB - 2 ) . There is no significant between the groups and layers. To use Western Blot detected the expression of MMP -2,TIMP -2 and MTl - MMP in 30 breast cancer cases and 30 breast innocent tumor cases, the expression of MTl - MMP in breast cancer and breast innocent tumor respectively is 90. 50 ±27.71,23.70 ±9. 35(t= - 12.51 ,P <0. 001);the expression of MMP -2 in breast cancer and breast innocent tumor respectively is 140.23 ± 46. 82,47. 30 ± 16. 03 (t = - 9. 35 ,P < 0. 001);the expression of TIMP - 2 in breast cancer and breast innocent tumor respectively is 67. 57 ±37. 10,10.40 ± 7.44(t= -9. 30,P <0. 001) , there is a significant difference between the expression of MMP - 2, TIMP - 2 and MTl - MMP in breast cancer and that in breast innocent tumor. And also this experiment is intended to discover the relationships of the protein expression of MMP -2, TIMP -2 and MTl - MMP in breast cancer, the sample size, however, is a little small, so it can not be able to support dependability analysis, therefore the conclusion of three markers dependability was not made. And next through the Western Blot method, to determine the expression of MMP - 2, TIMP - 2 and MTl - MMP in breast cancer was divided into groups and layers by the clinical patho -factors of breast cancer( T stage, N stage, TNM stage, ER, PR and C — erbB - 2 ) . Absolutely there is a significant difference between II stage and M stage in the expression of MMP -2 and TIMP - 2 in breast cancer. The expression of MMP - 2 in II stage and IH stage is 160. 38 ± 47. 16,124. 80 ± 27. 93 (t = 2. 25 , P = 0. 0356) respectively;The expression of TIMP -2 in H stage and H stage is 53. 38 ± 17. 02,80.40 ± 38.49(t = - 2. 27,P = 0. 0338) respectively.ConclusionsTo sum up, there is a significant difference between the expression of MMP - 2, TIMP - 2 and MTl - MMP in breast cancer and that in breast innocent tumor under the method of Western Blot, that is to say that the associated detection of MMP - 2, TIMP - 2 and MTl - MMP can hint the incidence of breastcancer. And also there is a significant difference between E stage and M stage in the expression of MMP - 2 and TIMP - 2 in breast cancer, it means that MMP - 2 and TIMP - 2 can have the contribution to hint and instruct for II stage and HI stage breast cancer patients in tumor malignant behavior, prognosis and treatment , meanwhile they can also reflect the tumor has the higher malignant biological behavior and it is possible to become a target of breast cancer bad prognosis. To use Immunohistochemistry method, we can detect that Positive expression is mainly locate in the cancer cell intracytoplasm, to show diffusive distribution of brown granulo, and negative expression is in the cancer cell nucleus, as well as less expression is in the interstitial cell. Meanwhile the significance of negative findings was not denied yet, there is no significant difference between the positive and negative group of ER, PR and C - erbB - 2 for the expression of MMP - 2, TIMP - 2 and MT1 - MMP in breast cancer. |
PDF Full Text Request