Inhibition Of RNAi On Expression Of EGFR In Non Small-Cell Lung Cancer Cells

Objective: To construct four plasmids which can inhibit expression of EGFR of NSCLC cell line A549 and observe inhibition ratio of EGFR expression and cell proliferation for the purpose to explore a new treatment of cancer .Methods: 1. Four short hairpin RNA (shRNA) were designed according to the homo sapiens EGFR mRNA ID, chemically synthesized, and reconstructed to pSilencei^? 2.1-U6 hygro plasmids. 2. Four recombinent plasmids and control plasmid were transfected to A549 cells, respectively. 3. The inhibition ratios of cell proliferation were detected by MTT. 4. With β-actin as inside control, the mRNA level of EGFR was detected by RT-PCR.Results: 1. Four plasmids containing shRNA were constracted successfully. 2. Four recombinent plasmids and control plasmid were successfully transfected to A549 cells and Hygromycin B was used to gain the cell clones that express stably. 3. The highest inhibition ratio of A549 cell proliferation was 88.85% .4. Compared with the blank control group, the mRNA level of EGFR was suppressed to 71.2% by RT-PCR.Conclusion: The recombinent plasmids can efficiently express EGFR specific shRNA, decrease mRNA level of EGFR expression and inhibit A549 cell proliferation.