The Effect Of HARD1Which Is After RNA Interference On Proliferation Of Colorectal Cancer Cell Lines

Objective:The experiment is working on the effect of hARD1which is after RNA interference on the growth and proliferation of Colorectal Cancer cell lines, and providing a theorical basement that hARD1can be a tumor marker in the diagnosis and treatment of Colorectal Cancer.Methods:The experiment use SW620、HCT-8、LS-174T as study objects of Colorectal Cancer cell lines, and filter the siRNA with effectively silent hARD1to transfect Human Colorectal Cancer cell lines. Through the adoption of EdU to analysis the effect of hARD1on the biological behavior of Colorectal Cancer cell lines.Steps:1.Culture cells of SW620、HCT-8and LS-174T.2. Design and synthesis siRNA.3.Use siRNA to transfect LS-174T、SW620and HCT-8cells.4. Extract and check total RNA.5. Use RT-PCR and Western Blot to check the effect of transfection.6. Use EdU to check the proliferation of cells.Results:1. HCT-8and SW620in vitro are cultured and had a good growing condition.2. Transfect siRNA.3. The QC Results of total RNA samples are28S/18S>0.7.4. Use β-actin and GAPDH as Analysis Internal reference to analysis siRNA, and the results are basically the same. 5. After24h transfection,48h tramstection,72h transfection, using EdU and Hoechst double staining to check proportion, suggesting the proportion rate of transfection cells is lower than the negative control group and blank control group.Conclusion:1. Establish three Human Colorectal Cancer cell lines which transfected by siRNA-ARD1.2. The proliferation of three human Colorectal Adenocarcinoma cells decrease after hARD1being siRNA interferenced. Speculate that hARD1can be used as one of the targets of clinical treatment.3. LipofectamineTM RNAiMAX Liposomes can effect the proportion of cell lines, and participate in biology activities.