| Objective:Renal cell carcinoma (RCC) is the most common tumor arising from the cells in the lining of tubules in the kidney. It is characterized by a lack of early-warning signs, which results in a high proportion of patients with metastases. Median survival for patients with metastatic disease is about13months. So there is a great need to find a new target in the inhibition of RCC proliferation.Hepatocyte growth factor (HGF) is a heterodimers molecule with a number of biological activities, including regulation of migration, invasion and angiogenesis in cancer. Over expression of HGF and its receptor. c-Met, in renal cell carcinoma have been reported. Moreover, a higher plasma level of HGF in RCC patients is associated with an advanced stage of malignancy and a poor prognosis.Transient receptor potential (TRP) channels are a large family of nonselective cation channels. TRPC channels, a subfamily of TRP channels in mammalian cells, include7members (TRPC1～7), several studies have shown that TRPC6channels were involved in cell proliferation including different types of cancer cells.Recently, it has been shown that TRPC6is critical for HGF-induced growth, migration and morphogenesis in human renal tubular cell line HK2. Additionally, in Huh-7cells, the expression of EGF and HGF receptors in hepatocarcinoma probably induces TRPC6expression, increasing the rate of cell proliferation. Furthermore, study was designed to investigate the possible relationship between TRPC6and HGF and the function of TRPC6channels in cell proliferation of prostate cancer by using physiological and molecular techniques.MethodsA. Molecular (RT-PCR and Western blot), and immunohistochemical techniques were used to investigate TRPC6expression.B. To inhibit TRPC6activity or expression, RNA interference was used.C. The effects of TRPC6channels on RCC cell proliferation, and cell migration were investigated by MTT, and scratch assay. D. Flow cytometry was designed to clarify the cell cycle progression.ResultsA. TRPC6was expressed in1932and ACHN cells.B. Expression of TRPC6was markedly increased in RCC specimens than that in normal renal tissues; moreover the level of TRPC6expression was associated with RCC histological grade.C. Western blot revealed a decrease of TRPC6protein expression in the TRPC6siRNA groups. Cell proliferation analysis showed that the number of ACHN cells transfected with siRNA3was markedly reduced compared with the control cells or the cells infected with siRNAc. Moreover TRPC6knockdown markedly reduced the ability of HGF-induced ACHN cells proliferation.D. There were no differences of migration rate between the control group and the siRNAc group. However, the migration of the siRNA3group was significantly suppressed, and the gene silence of TRPC6markedly reduced the ability of HGF to increase cell migration.E. Blockade of TRPC6channels arrested RCC cells at the G2/M phase of the cell cycle.ConclusionsA. TRPC6was highly expressed in ACHN cells and1932cells, and expression level was associated with RCC grade.B. Inhibition of TRPC6expression suppressed ACHN cells proliferation and migration in vitro. TRPC6channel is critical for HGF-induced proliferation and migration.C. Inhibition of TRPC6expression induced G2phase arrest of the RCC cells cycle.D. These results suggest that TRPC6channels contribute to RCC development and essential for the HGF-induced cell proliferation and migration of ACHN human RCC cells. Thus we suggest that TRPC6may be a novel target for therapeutic intervention against RCC... |
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