| Background Esophageal carcinoma is a common malignant tumor of digestive tract, its morbidity ranks the eighth of malignant tumor in the world and the fourth in China. Although people has made a long-term study of the treatment plan including many methods like operation resection, chemotherapy radiation therapy and so on, the survival rate of 5 years is only 15%. Many reasons lead the high mortality after the resection of esophageal carcinoma, postoperative metastasis is an important reason.Urokinase plasminogen activator (uPA)can activate the plasminogen and a variety of other proteolytic enzymes. They also have a positive role in the invasion and metastasis of tumor, because they can degradate the natural barrier of extracellular matrix which impede cell metastasis(extracellular matrix,ECM), and clear the resistance for tumor cells when they transfer. Matrix metalloproteinases(MMPs) has an enzmy system that can degradate the ECM and destroy the integrity of basement membrane(BM), they also play a very important role in the transfer of tumor cells. Experiments have confirmed that hUCMSCs lysate can inhibit the proliferation and migration of esophageal carcinoma EC9706 cells.Objective The objective is to investigate the effect of the lysate of hUCMSCs on the expression of uPA and MMP-2 in esophageal carcinoma EC9706 cells. This can establish an theoretical foundation for further research of the mechanism function of inhibition. It can also provide a theory basis for the clinical application of stem cell to the treatment of inhibiting the tumor metastasis of esophageal cancer.Methods 1.Culture the esophageal carcinoma EC9706 cells; 2.hUCMSCs was cultured by using tissue culture method, cell phenotype was analysed using the flow cytometry when cells experienced three times passage; 3. Collected the hUCMSCs and prepared hUCMSCs lysate using the repeated freezing and thawing method.4. Each group had the same number of esophageal carcinoma EC9706 cells, added different amount of hUCMSCs lysate to each group, then cultured for 60h.Texted the expression level of uPA and MMP-2 in EC9706 by Western-blot.Result The morphology of esophageal carcinoma EC9706 cells are polygonal and have a various of large nucleus-. irregular shape and irregular arrangement of cell populations checking with inverted microscope.When hUCMSCs was primary cultured 7-12d with the tissue culture method fusiform or polygonal fiber cells can be seen under the microscopically. Then we can found that hUCMSCs presented fibroblast and arranged in swirl shape. We analysed the third generation hUCMSCs with the flow cytometry and found that of the MSC smarker CD90、CD166 had an high expression, but the hematopoietic specific cell surface cell markers CD34、CD45 did not have an expression.After the lysate of human umbilical cord mesenchymal stem cells treated esophageal cancer EC9706 cells, the expression of uPA in the control was 0.28075±0.00233, 0.28790±0.00176 in the group of 1/2 times,0.13782±0.00542 in the grop of 1 times, 0.12465±0.01277 in the grop of 2 times. Expression of MMP-2 in the control was 0.27051±0.01956,0.26698±0.00231 in the group of 1/2 times,0.13017±0.01529 in the grop of 1 times,0.11755±0.00982 in the grop of 2 times. uPA level and MMP-2 level in EC9706 did not change when the lysate from hUCMSCs was less than EC9706 cells in the treatment compared with untreated control cells (P>0.05). The expression of uPA and MMP-2 in esophageal carcinoma cells was decreased significantly when the lysate from huCMSCs with cell numbers equal or greater than treated EC9706 cells (P<0.05)Conclusion Human umbilical cord mesenchymal stem cells lysate inhibited the migration of esophageal carcinoma EC9706 cells which may be associated with uPA and MMP-2. |
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