The Efficacy Of Vector-based RNA Interference Technology To Inhibit The Proliferation Of Non-small Cell Lung Cancer Line A549

Objective: Constructing plasmids vector expressing small interfering RNA (siRNA) to investigate whether the vector-based RNA interference technology inhibit the proliferation of the non-small cell lung cancer cell line A549. Methods: 4 plasmids expressing specific siRNA targeting epidermal growth factor receptor (EGFR) gene were constructed to transfect the A549 cells mediated by liposome Lipofectamine^?2000. 7 groups were divided for transient transfection (pSilencer-1,pSilencer-2, pSilencer-3, pSilencer-4, p53-pcDNA, liposome and control group) .MTT method was used to measure the growth of the cells at day 1 , 3, 5, 7 after transfection ,and to calculate the inhibition ratio of each group .The cells stably transfected with 4 recombinant pSilencer^?2.1-U6 hygro plasmids were screened by Hygromycin . To detected the cell cycle through the flow cytomettry and compare the G0/G1 phase of each group .Results: 1. Plasmids expressing specific siRNA targeting EGFR gene were constructed successfully , and were certified through agarose gel electrophoresis and gene sequencing .2.The proliferation of A549 cells decreased after transient transfected by pSilencer-1, pSilencer-2, pSilencer-3 and pSilencer-4 comparing to the liposome group and control group (p<0.05),while there was no difference between the 4 groups and the positive control P53-pcDNA group (p>0.05).There was no difference between the liposome group and the control group (p> 0.05).The 4 recombinant plasmids had the similar efficacy of inhibition and it didn't change with time (p>0.05). 3.The pSilencer-1, pSilencer-2, pSilencer-3 , pSilencer-4 and the P53-pcDNA group had the higher inhibition ratio than the...