The Role Of NEDD9in Human Hepatoma Cell Proliferation And Migration

Background:Hepatocellular carcinoma is a malignant tumor which has high incidence, high degree of malignancy,easy metastasis and development in the world. Metastasis and development are the main cause for the deathof patients with liver cancer. However, there has not been reliable methods for treatment.This forcesclinicians try to explore the genes change related to the metastasis and development of liver cancer.Scientists hope to clarify the mechanism of liver cancer development and metastasis at the molecular levelin order to solve the problem of liver cancer recurrence and metastasis fundamentally. NEDD9is Neuralprecursor cell Expressed Developmentally Down-regulated9，which may be involved in development andmetastasis in hepatocellular carcinoma.Objective:Detect the expression of NEDD9in hepatocellular carcinoma cell lines. Express different domains ofNEDD9fused with GST in the prokaryotic cell(E.coli).Observe the change of the cells migration,proliferation and apoptosis with NEDD9knock-down using RNAi interference.Methods:1. Etract the total RNA.Detect the nucleic acid expression of NEDD9and a few pieces of functionaldomains in SMMC-7721cell line.2. Express NEDD9functional domains with GST in E.coli for further investigation of NED9biological function.3.Select the interference target of NEDD9..Design and synthesis oligonucleotides which is clonedinto pSilencer3.1,after annealing into double stranded construction. Transfect the recombinant vector intoSMMC-7721which is derived from liver cancer cells.4.Use the scratch test to detect the change of the cells migration ability.5.Detect the change of the cells proliferation by cell counter.6.Detect the change of the cells vitality by MTS. Results:1.The NEDD9is highly expressed in SMMC-7721cells.2.Constructed recombinant vector that would overexpress NEDD9in SMMC-7721cells line.3.ShRNA expression vector for NEDD9was successfully constructed.After transfection, the NEDD9expression in mRNA level decreased significantly.4.The lower NEDD9expression in SMMC-7721can inhibit the migration of the cells withtransfection.5.The lower NEDD9expression in SMMC-7721can inhibit the proliferation of the cells withtransfection.6.The lower NEDD9expression in SMMC-7721can inhibit the vitality of the cells.Conclusions:This study suggests that NEDD9is highly expressed in HCC cell lines. The experiments show itplays an important role in the proliferation metastasis and recurrence of liver cancer.It is a molecule relatedto the metastasis and recurrence of liver cancer cell line. It is expected to become a marker of liver cancerinvasion and metastasis or a treatment target against the metastasis and recurrence of liver cancer inclinical practice..