Effect Of Hepatocarcinoma Cell Proliferation, Migration And Proliferation Under The Down-regulation MTDH Gene Expression Through MTDH-siRNA

Background:Hepatocellular carcinoma(HCC) is one of clinical common malignant tumor, its incidence rate is increasing year by year, according to the report, the global annual increase of about 500000 liver cancer patients, about half of which in our country, a high mortality rate of malignant tumors in China second. The treatment of liver cancer is limited, liver resection and liver transplantation is the best choice of potential, but only 10%-20% of patients have conditions of treatment in patients; With advanced hepatocellular carcinoma patients cannot operation excision, minimally invasive treatment is to a certain extent, local tumor control lesion growth, relieve local symptoms but not cure cancer, most HCC patients eventually died of tumor invasion and metastasis.. Therefore, the urgent need to find a safe, more effective, low toxic treatment method. The recent studies have shown that MTDH/AEG-1 has a cancer gene function, can promote tumor cell proliferation, invasion and metastasis of tumor, increase resistance, the main processes involved in tumor progression in many malignant tumor tissues. Compared with benign hepatic tumor and normal liver tissue,expression of MTDH/AEG-1 in hepatocellular carcinoma tissue and cultured hepatoma cells increased significantly. This thesis proposes to set up using Hep G2 cell line, by small interfering RNA Technology(si RNA small interfering RNA,) down regulated the expression of MTDH, and observe its effects on the biological behavior of hepatocellular carcinoma cell proliferation, migration, apoptosis and so on, through the above study to verify the targeting of MTDH/AEG-1 is effective in the prevention and inhibition of liver cancer growth, metastasis.Objective: Studying the interference by MTDH gene expression of Hep G2 cell proliferation, migration and apoptosis of liver cancer.Methods: Using si RNA technology, lower Hep G2 liver cancer cell MTDH gene expression, immunocytochemistry and Western bolt experiment testing interference effect. Determined by MTT test MTDH genes influence on Hep G2 cell proliferation, TUNEL apoptosis experimental detection of cell apoptosis, flow cytometry(FCM) to detect the cell cycle, scratches the experimental cell migration ability, rt-pcr detection the Bcl- 2, Bax m RNA expression validation promote cell apoptosis.Results: Through the MTDH- si RNA interference MTDH gene expression, liver Hep G2 cell proliferation capacity decreased obviously, 48 hours and 96 hours of proliferation inhibition rate was 43.8% and 64.6%, respectively; Cells obviously block in the stationary phase(G0) or early(G1) DNA synthesis, cell apoptosis rate increased, the apoptosis rate increased by 23.46%, significantly lower cell migration ability, Bax gene expression increase obviously, and cut the Bcl- 2 expression, promote cell apoptosis.Conclusions: MTDH- si RNA interference by MTDH gene expression inhibits Hep G2 cell proliferation, block in the G0 / G1 phase, and inducing cell apoptosis, can obviously inhibit Hep G2 liver cancer cell migration ability. MTDH gene is expected to become the new target for the treatment of liver cancer.