In Vitro Experimental Study On Anti-fak Short Hairpin RNA For Metastasis Of Colorectal Cancer

Background and ObjectivesColorectal cancer is one of the common malignant tumor of digestive tract,whose incidence increased year by year. At present,surgery is still the main treatment of colorectal cancer,supplemented by radiotherapy and chemotherapy combined treatment in the same time,But for the people who has been in the later period and who has been tranfered,The clinical outcomes was poor. The death of patients with colorectal cancer is mainly caused by tumor metastasis. Tumor metastasis is a complex process.The tumor cell invade and break through the extracellular matrix is the basic steps,then enter blood,Reach distant organs through the blood,break blood vessels into the target organ for settlement and proliferation.Studies shown that:focal adhesion kinase as an important intracellular signaling molecule,mediating intracellular signaling network of cross-linking,who play a key role in cells'movement and invasion.The expression of FAK in primary cancer and liver metastases more than in normal colorectal mucosa.Therefore, Interventing the expression of FAK could provide a new way to treat the colorectal cancer. RNA interference(RNAi) has a strong role in inhibition of the targeted gene,which is more than Antisense oligonucleotides.RNAi is considered as a landmark after the PCR method of genetic engineering.Methods1.To inhibit the expression of FAK in colon carcinoma SW620 cells by generation of RNA interference(RNAi)construct targeting FAK.2. Expression of FAK were detected by RT-PCR and Western blot. MTT was employed to detect the effect of FAK silence on the proliferation of SW620 cells.3. Prepare Transwell chambers to detect the effect of FAK silencing on invasion and metastasis of SW620 cells. Results1. The targeting FAK RNAi lentiviral vector was constructed successfully.2. The expression of FAK mRNA and protein level was found to reduce by RT-PCR and Western blot in interfere SW620 cells compared with untreated group and negative plasmid control group,respectively. MTT test demonstrated that growth of the cells was inhibited in interference group, but not in untreated group and negative control group (P <0.05).3.Invasion in vitro experiments: To compare the untreated group and negative control group cells, the ability of cell invasion and metastasis decreased significantly in the treated group (P <0.05), the normal group and negative control group had no significant difference (P>0.05)Conclusions1. Constructing successfully FAK RNAi lentiviral vector.2. Reconstructing lentiviral vector effectively inhibited colorectal cancer cells SW620 transfected with proliferative activity.3.The colorectal cancer cells SW620 invasion and metastasis were significantly inhibited by FAK silencing.