The Role Of CK147and MMPs In The Invasion And Metastesis Of Hepatoma

CD 147 is a highly glycosylated transmembrane molecule belonging to the immunoglobulin (Ig) superfamily and is involved in cell-cell adhesion .It is present on the tumor cell surface.Peritumor fibroblast are stimulated by these CD 147 at the surface of cancer cells to secrete increased amounts of MMPs .Our aim is to study the function of CD 147 and MMPs in the invasion and metastesis of cancer cells from tumor cells and their circumstance. I The expression and relationship of CD147 and MMPs in human hepatoma cell lines. Methods: The expression of CD 147 in human liver cell QZG and different hepatoma cell lines (BEL-7402 QGY-7404~ SMMC-772 1 HHCC IIHCC-92 04) was investigated by flow cytometer. ZymograPlI was used to study the content and types of positive MMPs. The expression of CD 147 and MMP-2 in human hepatoma cell line HHCC was observed by using immunohistochemical ABC method and laser scanning confocal microscopy. Results: CD 147 was positively presented in five hepatoma cell lines , but in normal liver cell was negative MMP-2 and MMP-9 were detected by all the hepatoma cell lines and normal liver cell but the expression in hepatoma cell lines was obviously high. The relation between CD 147 molecule and the expression of MMPs was positive relation. CD 147 immunoreactivity was localized in the membrane and cytoplasm of tumor cells, while MMP-2 immunoreactivity was existed in the cytoplasm of tumor cells. Conclusion: Hepatoma cell can secrete MMps by itself.CD147 could be used as a marker to judge the invasion and metastasis of hepatocellular carcinoma. II Affinity purification and functional identification of hepatoma membrane associated antigen IIAb18G Methods: An affinity chromatograPHic column was prepared by coupling mAb HAbI8 to CNBr-activated SePHarose-4B. HAb18G was obtained by affinity chromatograPHie purification from homogenate of fresh surgical hepatoma specimen. The purity and specificity of HAbJ 80 was tested by high performance capillary electroPlioresis and Western blotting respectively. The function that HAb 180 stimulates fibroblast to Secrete MMPs was identified by zymograPHy. Results: The purity rate of HAb18G was 96% after affinity chromatograPHy; HAbJ 8G reacted with mAb HAb1 8 specifically ;zymograPHy showed that the quantities and kinds of MMPs stimulated by HAbI 8G were different from those secreted only by fibroblast. Conclusion: HAb18G could be purified from an affinity chromatograPHic column coupling mAb HAb 18 to CNBr- activated SePHarose-4B; This hepatocarcinoma membrane antigen flAb 1 8G has the same function as the Extracellular Matrix Metalloproteinase Inducers (EMMPRI7N/CD 147)...