The Effects Of HGF On The Proliferation And Migration Of ECV-304, HDLEC, SW480 And SW620

Colorectal carcinoma is the second common malignant tumor in digestive tract. The main reason for its treatment failure is the development of hematogenous metastasis and lymphatic metastasis. It is shown that both the invasive capability of cancerous cells and the neogenesis of blood vessels and lympgatic vessels are closely related with the metastasis of colorectal carcinoma. And the density of blood vessels and lympgatic vessels is correctly related with lymphatic metastasis and the prognosis of colorectal cancer. Inhibiting the neogenesis of blood vessels can decrease the blood supply of the tumor, induce the apoptosis of cancerous cells and reduce the incidence of metastasis. The effective treatment target, especially hepatocyte growth factor (HGF), for colorectal cancer has been widely studied.HGF, also known as scatter factor (SF), was first identified as a mediator of liver generation, but it is also a growth factor with pleiotropic effects including mitogen, mobility factor and morphogen and so on. HGF's activities are mediated by a membrane-spanning receptor tyrosine kinase, the HGF receptor (HGF-R, also known as MET/c-met). It has been proved that HGF plays an important role in the infiltration and metastasis of cancerous cells and the neogenesis of blood vessels. Recently, many studies indicate that HGF may be an enhancing factor in the neogenesis of lymphatic vessels. Moreover, angiogenesis and lymphangiogenesis are directly related with the infiltration and metastasis of carcinoma. The angiogenesis and lymphangiogenesis is a process of proliferation, migration and canaliculization of the endothelial cells. However, the role of HGF in the the neogenesis of lympgatic vessels it is still unclear and it is short of systematic study of the role of HGF in the angiogenesis and lymphangiogenesis of colorectal carcinoma.Vascular endothelial growth factor(VEGF)-A have been identified as one of the specific angiogenesis factors, while vascular endothelial growth factor (VEGF)-C and -D play important roles in lymphangiogenesis and metastasis. Therefore, to explore how the cancer cells control the birth of VEGF and then promote the metastasis of the cancer cells while inducing the angiogenesis and lymphangiogenesis becomes a key issue in the research. It has been proved that HGF can increase the BEGF-A expression of colorectal carcinoma, but whether it can adjust the expression of VEGF family has not been systematically studied.This experiment uses a combinative method of form and function to examine systematically the effects of HGF on the proliferation and migration of ECV-304, HDLEC, SW480 and SW620 and the influence of HGF on the expression of FEGF-D in SW480 and SW620, in the hope of finding a new therapeutic target for restraining the neogenesis of blood vessels and lymphatic vessels of colorectal carcinoma.The present study was devided into two parts.1 The effects of HGF on the proliferation and migration of ECV-304 and human dermic lympgatic vessel endothelial cells (HDLEC).The effect of HGF on the proliferation of endothelial cells was examined by MTT and the method Flow Cytometry (FCM) was used to test the effect of HGF on the cell cycle of endothelial cells. And the ultrastucture of endothelial cells treated with HGF was shown by the transmission electron microscope (TEM).HGF could promote the proliferation of ECV-304 in a concentration-dependent manner. But the effect of HGF on the proliferation of HDLEC didn't increase with the concentration of HGF. The results of FCM showed that the cells of HGF group in G0 /G1 phase decreased obviously, but the cells in S phase and G2/M phase increased. The proliferation index of ECV-304 increased compared with the control group ( 47.5% vs 37.8%) and the proliferation index of HDLEC increased compared with the control group (37.1% vs 27.3%), which suggested that HGF mainly induced the cells to enter M phase. HGF could promote the migration of ECV-304 and HDLEC. HGF (>5ng/ml) obviously promoted the migration of ECV-304 in a concentration-dependent manner. Similarly, HGF effectively promoted the migration of HDLEC in a concentration-dependent manner. As shown by TEM, the microfilament of ECV-304 and HDLEC after being treated with HGF increased remarkably.2 The effects of HGF on the proliferation and migration of SW480 and SW620 and on the expressions of VEGF-A, VEGF-C and VEGF-D.MTT was used to analyse the effect of HGF on the proliferation and the effect of HGF on the cell cycle was determined by FCM. The effect of HGF on the migration of cells was studied by restoration of basal membrane; the ultrastructure of SW480 and SW620 after being treated with HGF was observed by TEM; the expressions of VEGF-A, VEGF-C and VEGF-D were determined by Western Blotting.HGF (≥10ng/ml) could promoted the proliferation of SW480 and SW620. It promoted the proliferation of SW620 in a concentration-dependent manner but in a very feeble way. The effect of HGF on the proliferation of SW480 didn't increase with the concentration of HGF. The results of FCM showed that the SW480 cells of HGF group in G0 /G1 phase decreased, but the cells in S phase and G2/M phase increased compared with the control group. However, there are weak changes in the SW620 cells of HGF group. The proliferation index of SW480 increased compared with that of the control group (46.9% vs 42.3%), and the proliferation index of SW620 increased compared with that of control group (33.8% vs 32.0%). HGF could promote the migration of SW480 and SW620. HGF promoted the migration of SW480 and SW620 in a concentration-dependent manner. As shown by TEM, the microfilaments of SW480 and SW620 increased remarkably after being treated with HGF. HGF upregulated the expressions of VEGF-A, VEGF-C and VEGF-D of SW480 and SW620 repectively.Conclusion1 HGF can promote the proliferation and migration of ECV-304 and HDLEC, and HGF can promote the migration of ECV-304 and HDLEC in a concentration-dependent manner.2 HGF can promote the proliferation and migration of SW480 and SW620, and HGF can promote the migration of SW480 and SW620 in a concentration-dependent way.3 HGF can promote the expressions of VEGF-A, VEGF-C and VEGF-D of SW480 and SW620 repectively.