The Experimental Study Of Human Lung Cancer-associated Gene By RNA Interference

Objective: To study the effect of pointmutational PP2A α gene on the expression of N35 which is the protein corresponding to monoclonal antibody N-35 , and to find the proof that pointmutational PP2A α gene is employed as the candidate target gene of gene therapy of lung cancer. Methods: GLC-82 cell lines were used as the target cells, and pointmutational PP2A α gene was used as the target gene. ShRNA plasmid expression vectors were constructed and transfected into GLC-82 with the cationic lipid .Positive clones werescreened out by G418. The difference of N35 expression was detected by Western Blotting.Results: All of shRNA plasmid expression vectors were successfully introduced into cultured GLO82 cell lines and could make those cells grow well in the medium containing G418 during a quite long period. GLC-82 cell lines transfected with pGenesil-1 -ZZS4 had a reduction of N35 expression compared with the non-transfected ones,so did those transfected with pGenesil-l-ZZS2. But the former was more significant than the latter.Conclusions: RNAi is an effective and specific gene silencing technology. ShRNA plasmid expression vectors can mediate RNAi and specifically suppress the target gene expression. Inhibiting pointmutational PP2Aa gene can lead to a reduction of N35 expression , and it is an fairly important Human lung cancer-associated gene. It is greatly reasonable that pointmutational PP2Aa gene is employed as the candidate target gene of gene therapy of lung cancer.