Effects Of Inhibition Of Methyl-CpG-binding Protein 2 By Sh-RNA On Proliferation And Apoptosis Of Lung Carcinoma Cell A-549

Objective : To construct the eukaryotic expression plasmid of short hairpin RNA(shRNA)targeted against MeCP2 gene(methyl-CpG-binding protein 2) and investi--gate the effects of MeCP2 inhibited by shRNA on the proliferation and apoptosisof lung cancer cell lines A-549.Methods : Finding the sequence of human MeCP2 gene in Pubmed,the oligonucleotides of shRNA were desingned by the software of ambion company, then sythesized and directionlly cloned into plasmid pSilencer-2.1-U6 with Amp gene and Hygro gene.The recombinant vectors were comfirmed by PCR and DNA sequencing analysis. The recombinant vectors were transfected into A-549 cell line by lipofectamineTM 2000,the effects on MeCP2 at mRNA and protein levels were observed by RT-PCR and Western blot,the changes of cell cycle and cell proliferation were observed by flow cytometry analysis and MTT.Result:Two shRNA expressing vectors were recombined and transfected into A-549 cell line successfullly.The mRNA and protein of MeCP2 were reduced visviblly.The down regulation of MeCP2 increased the apotosis percentage of A-549 cell and decreased the growth speed of A-549 cell.Conclusion: The MeCP2 gene is proved to have promote the growth of A-549 lung cancer cells. RNA interference the expressionof MeCP2 can increase the apotosis of lung carcinoma cells.