The Initial Study About The Functionary Of HPA In The Colorectal Cancer Metastasis

Objective : Colorectal cancer is one of the common gastrointestinal neoplasms, and its incidence rate is rising year by year. Although the different available therapies, which including surgical operation ,radical therapy, and chemical therapy , have currently been used in the treatment of colorectal cancer, and all of these have markedly improved the prognosis of colorectal cancer. But results are not satisfied. As for the patients visiting, about 15%～35% already have tumor metastasis because there are not overt symptoms in the early period of colorectal cancer, and tumor metastasis is one of the majority reasons to result in death .At the end of 70s, it is found that the metastasis of tumor was relativity with a sore of substance like heparin. Although these phenomena well-documented, it has takes 20years to isolate a heparanase gene, mainly because of instability in designing specific, quantitative assays. Cloning and functional characterization of the long sought-after heparanase opens a new chapter in the understanding and potential manipulation of metastasis processes.Tumor invasion and metastasis have to break down the barriers of the extra cellular matrix (ECM) and blood vessel basement membranes (BM).Heparan sulphate proteoglycans (HSPG) localized in the extracellular matrix and on the external surface of cell membranes, play a major role in cell-cell and cell-extracellular matrix interactions. It is the important constitute parts of basement membranes. Heparanase is an endo-β-glucuronisase which is one of the only found to degrade HSPG, It can cleaves heparin sulfate (HS) to degrade ECM and BM ,and can facilitate to release or activate the HSPG-bounded growth factors(such as bFGF, VFGF et al.) to induce angiogenesis. In this way,HPA promotes tumor invation and metastasis . It is becoming a new target to treat cancer with the development of the study about the inhibitor of HPA and the relationship between the HPA and the tumor invasion and metastasis.To investigate the relationship between HPA and the metastasis of colorectal cancer,we detected the expression of HPA in normal colorectal mucosa, primary colorectal carcinoma ,lymph node metastasis and liver metastasis by using semi-quantitative RT-PCR and immunohistochemical method, then analyzed the relationship between HPA and the clinicopathoplogic features of colorectal cancer.Methods: 1 Immunohistochemistry was performed by using the SP method. We use anti-HPA (1:60 dilution) to detect the expression of HPA protein in samples, then analyzed the results with the clincopathologic features .2 Semi-quantitative RT-PCR: The present study was proformed to detect the expression of HPAmRNA andβ-action by using RT-PCR and measured the expression level of HPAmRNA, then analyzed the results.Results: By immunohistochemistry analysis on samples from patients with colorectal cancer, the positive expression rate of HPA were 18.60% (8/43), 68.75% (33/48), 37.5% (3/8), 82.20% (37/45) In normal intestinal mucosa, primary colorectal cancer, normal lymph nodes, metastatic lymph nodes. In the 48cases , there were 9 cases with liver metastasis, 13 cases were final diagnosed to be poorly differentiated adenocarcinoma, 13 cases were moderately differentiated adenocarcinoma, 22 cases were well-differentiated adenocarcinoma. In 41cases with chorion invaded , the positive expression was 31cases (75.61%). In 7 cases without chorion invaded , the positive expression was2cases (28.57%). The expression of HPA protin in moderately differentiated adenocarcinoma, well-differentiated adenocarcinoma and poorly differentiated adenocarcinoma,were 45.45% ,76.92%and 53.85%. in 9cases with liver metastasis, the expression of HPA protein were all detected .By statistic analysis, HPA expression level was higher in primary colorectal cancer than it in normal intestinal mucosa (χ2 =23.04 P<0.05), was higher in colorectal cancer with metastasis lymph nodes than it in colorectal cancer without metastasis (P<0.05), was higher in colorectal cancer ofⅢ～Ⅳ(TNM staging) than it inⅠ～Ⅱ(P<0.05), and was higher in cancer (Dukes D and TNMⅣstage) than it in other stages (P<0.05). Moreover there was statistic significance in all these results. The analysis on clinical data showed that HPA expression in primary colorectal cancer has no relationship with patient's gender, age, size and differentiated level (P>0.05). In cases with placenta percreta invaded the expression of HPA protein was higher than that without chorion invaded(P>0.05).In 40 patients with colorectal cancer, the expression rate of HPA mRNA were5%(2/40),13.33%(2/15), 75%(30/40) and 78.13%(25/32) in intestinal mucosa, lymph nodes without metastasis ,primary colorectal cancer and metastatic lymph nodes by RT-PCR. In the 40 cases , there were 7 cases with liver metastasis, 13 cases were final diagnosed to be poorly differentiated adenocarcinoma,13 cases were moderately differ- entiated adenocarcinoma, 14 cases were well-differentiated adenocarcinoma. In 31cases with chorion invaded , the positive expression was 28cases (90.32%). In 9 cases without chorion invaded , the positive expression was 3 cases (33.33%). The expression of HPA protin in moderately differentiated adenocarcinoma ,well-differentiated adenocarcinoma and poorly differentiated adenocarcinoma,were45.45% ,76.92%and 53.85%. in 7cases with liver metastasis, the expression of HPA protein was 85.71%(6/7).Meanwhile it was higher evidently in colorectal cancer with liver metastasis than it in cancer without metastasis(P<0.05), was higher in group with metastatic lymph nodes than it without metastatic lymph nodes(P<0.05), was higher in metastatic lymph nodes than it in normal lymph nodes(P<0.05), and was higher in cancer of stageⅢ～Ⅳ(TNM staging) than it in cancer of stageⅠ～Ⅱ(P<0.05); In cases with placenta percreta invaded the expression of HPA protein was higher than that without chorion invaded.Conclusions:1 The positive expression rate of HPA was higher in metastatic lymph nodes and colorectal cancer with liver metastasis. It suggested that HPA expression was related with metastasis of colorectal cancer.2 With the tumor invasive more depth , the expression of HPA protein was increased. The expression of HPA protein was correlated with development of colorectal cancer3 HPA expression level was higher evidently in primary colorectal cancer than it in normal intestinal mucosa. It indicated that HPA expression might be related with the genesis of colorectal cancer.4 The analysis on clinical data showed that HPA expression in primary colorectal cancer has no relationship with patient's gender, age, size and differentiated level.