Effect Of DLK1Gene Silence By Interference In Development Of Hepatic Carcinoma

Hepatocellular carcinoma is a global malignancy, which had an insidious onset. it is too late when patients were diagnosed and most of them had lost treatment opportunity. Therefore, we urgently need to find a more sensitive diagnostic indicators. The imprinted gene DLK1has been recognized as a cancer related gene since it highly expressed in HCC, but not expressed in normal liver cells. In this study, We are using gene silencing to detect the changes of DLK1’s expression and to explore its role in Hepatoma cells, peritumoral cirrhosis liver cells and normal liver cells.[Objective]Explore the effection about apoptosisand proliferation of RNA interference silent DLK1gene in normal liver cells, peritumoral cirrhotic liver cells and liver cancer cell,and which provided laboratory evidence for the early diagnosis and treatment of HCC.[Methods]1. DLK1siRNA were transfected into normal liver cells, peritumoral cirrhotic liver cells and liver cancer cells with Lipofectamine2000Transfection Reagent.Expression level of DLK1in this three cell lines was detected by RT-PCR.2. Using flow cytometry to detect3cells about apoptosis and their cell cycle distribution after RNA interference.3. Using MTT and cell colony assay to detect three cell lines about cell proliferation and growth curve after RNA interference.4. Using Western Blot to detect3cells after transfection about Changes of DLK1protein expression。 [Results]1. It was proved that expression of DLK1in normal liver cells, peritumoral cirrhotic liver cells and liver cancer cells could be downregulated by siRNA with RT-PCR.2. Three cell lines’s apoptosis rate were increased after RNA interference. apoptosis of hepatoma cells increased significantly, and their cell cycle arrest in the S phase; followed by peritumoral cirrhosis liver cells,but apoptosis of normal liver cells is not obviously, their cell cycle arrest in G0/G1phase.3. Proliferation rate of three cell lines was significantly lower after RNA interference, peritumoral cirrhosis liver cells are the most obvious, followed by liver cancer cells,and the proliferation rate of normal liver cells are no obvious changes.4. The DLK1protein expression of three cell lines after RNA interference is lowest than the control group and negative control group.[Conclusions]DLK1gene silencing by RNA interference can promote the apoptosis of peritumoral cirrhotic liver cells, normal liver cells and liver cancer cells and inhibit their proliferation.