| Hepatocellular carcinoma (HCC) is one of the most popular and important malignant neoplasm in China. To elucidate the molecular mechanism of the proliferation, differentiation and transforming of the liver cell is very helpful for the early diagnosis and efficient treatment of the HCC. Hepatocellular carcinoma has a grave prognosis, particularly in advanced stage, because it spreads throughout the liver via the intrahepatic portal vein. With the improvements in early diagnosis, evaluation of hepatic functional reserve, surgical techniques and perioperative management, long-term survival has been achieved in some patients who undergo surgical resection. However, even when curative resection is performed at a relatively early stage of disease, a considerable number of patients develop early intrahepatic and/or extrahepatic recurrence postoperatively. In addition, molecular factors related to the tumor progression and portal vein invasion of HCC have not been well characterized. Several clinicopathologic risk factors are associated with early tumor recurrence, particularly portal vein invasion. Some molecular factors are related to tumor recurrence after hepatic resection, including telomerase activity. However, molecular markers that predict early tumor recurrence, particularly for early-stage HCC, remain limited. Osteopontin (OPN) is the principal phosphorylated glycoprotein of bone and is expressed in a limited number of other tissues including dentine. It was showed that the SPP1 gene comprises 7 exons, 6 of which contain coding sequence. A highly informative short tandem repeat (STR) polymorphism located at the SPP1 locus showed no recombination with the autosomal dominant disorder dentinogenesis imperfecta type II. Osteopontinwas alternatively identified as Secreted phosphoprotein 1(SPP1), Bone Sialoprotein, Urinary stone protein, early T-lymphocyte activation 1(Eta1). Osteopontin is a 44 kDa glycophosphoprotein found in all body fluids, the extracellular matrix components, and the proteinaceous matrix of mineralized tissues Osteopontin, as an acidic, highly phosphorylated and glycosylated calcium-binding secretory protein, is expressed widely and has a diverse range of functions. These include cell adhesion and migration, immune and in.ammatory response, antiapoptosis, suppression of nitric oxide synthase, and bone calci.cation.1,2 Human OPN contains an integrin-binding RGDS domain. It binds to cells via the RGDS cell adhesion sequence that recognizes the (v(3 integrin and to extracellular matrix. The RGDS motif is found in proteins that play a role in cell adhesion and metastasis. Osteopontin is expressed in excessive amounts by many transformed cell lines, and this overexpression is correlated with metastatic potential in the murine model. Antisense OPN can decrease OPN levels and suppress metastatic potential. Osteopontin is overexpressed in most human carcinomas. Its overexpression is correlated with advanced stage in gastric carcinoma, but not in lung, breast, and esophageal carcinomas. Despite the strong association of OPN expression with tumor metastasis in vitro and in animal studies, the role of OPN in human carcinoma, including hepatocellular carcinoma, is less clear. In present study we used real time quantitively reverse transcription PCR method to analize the expression level of OPN in the HCC and intrahepatic cholangiocarcinoma (ICC). The other method to determined the m RNA level of the genes such as Northern blot, semi-quantive RT-PCR and dot blot could only semi-quantitively caculate the expression level of genes. Here we used the HCC and ICC and corresponding adjacent liver tissues specimens to evaluated the expression OPN expression. We also made the rat HCC model with DENA and determined the OPN expression dynamically to evaluate the putative value of OPN as the diagnosis molecular marker and the target of treatment in the occurrence and preogression of HCC.PART I Investigation of gene expression of OPN in human HCCThe real time RT-PCR was successfully used to det... |
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