A Study On Establishing A Mouse Model For Human Liver Cancer And It's Application

Liver cancer is a common malignant tumor.In a narrow sense,liver cancer indicates the Primary Liver Cancer(PLC),the most commonly known cancer in clinical practice.According to the pathological types,PLC can be classified into hepatocellular carcinoma,cholangiocarcinoma,and combined hepatocellular-cholangiocarcinoma.China has the most PLC patients in the world.There are no significant symptoms appeared in the early stage of cancer,and the speed of progression of cancer cells is fast.Therefore,the tumor develops very quickly,resulting in late diagnosis and high mortality rate.The World Health Organization has listed liver cancer as one of the ten most commonly malignant tumors.Multiple genes,proved by scientists,have mutated in liver cells to become cancer genes: mutated P53,mutated P21,N-ras,C-myc,TGF?,C-ets-2,EGFR,IGF-?,IGF-? variants,CSF-1 receptors,C-erb-2,CCND1,mdm2,MMP-2,ICAM-1,etc.It is obvious that by examining the expressions of cancer genes(or the degree of activation)such as microarray assay can help diagnose PLC.Similarly speaking,if one can prevent these cancer genes from expressing,development of cancer cells will be suppressed,which is the focus and direction of current research for cancer treatment.Sorafenib,one of multi-kinase inhibitors,is the sole lisenced small molecular targeted drug for liver cancer.It's antitumous effects have been investigated and confirmed on multiple solid tumors.A lot of studies have proved that sorafenib has a strong inhibition on replication of liver cancer cells.This article describes building a mouse model for liver cancer through establishing luciferase-liver cancer cell line.For the luminance from the metabolic substrate of the luciferase is proportional to the number of cells,we can evaluate the inhibitory effect of sorafenib on human liver cancer cells in mouse.The experimental design of this project is as follow.We firstly introduced Luciferases' and eGFP reporter gene into the lentiviral vector(pWPXLD),which was then transfected into the 293 T cells along with PsPAX2 and PMD.2G Lentiviral packaging plasmids for to produce lentivirus;Afterwards,the virus was then transduced into human liver cancer cells(Huh7 cells),establishing a luminescent liver cancer cell line.Transduction efficiency was determined by the percentage of GFP positive cells detected with flow cytometry.After purified these tranduced cells,we next Injected different quantities of cells into the spleen of NOD SCID IL2?-/-mice,and to explore the optimal condition for building a mouse model of human PLC.This study manifests the inhibitory effects of sorafenib applied over five weeks on human liver cancer cells in mouse model,through comparing the luminous efficiency of the negative control and the experimental group.These results provided the rationale of the novel liver cancer drug to enter the clinical stage.