[Construction of FasL siRNA expression vector and its expression in lung cancer cell line A549.].
BACKGROUND
Fas/FasL is a member of Tumor Necrosis Factor (TNF) super family, and related to tumor cell apoptosis. It is hypothesis by forward study that activated lymphocytes is more sensitive with Fas/FasL due to up-regulation of FasL expression, so it can be inverse killedly and cleared by tumor cell. The aim of this investigate is to study the fuction of FasL gene and gene therapy of lung cancer by to down-regulationg the FasL gene expression with a siRNA expression plasmid in lung cancer cell A549 as well as its inverse killing effect between activated T lymphocytes and lung cancer cell A549.


METHODS
Potential RNAi oligonucleotides of FasL was designed and synthesized according to appropriate web site. Then a FasL siRNA plasmid was constructed using a pGCsi-U6 vector.The plasmid was sequenced to confirm the inserted sequence. Western blot analysis was used to assess the levels of FasL proteins after the constructed plasmids have been transfected into A549 cells.


RESULTS
It was confirmed by sequencing that the plasmid was constructed successfully. The result of Western blot clearly showed that FasL siRNA plasmid inhibited FasL expression in A549 cells.


CONCLUSIONS
The construct of FasL siRNA plasmid is successful. FasL protein expression of A549 cell is effectively inhibited by RNAi .
Wang
Haidong
H
Cardiothoracic Surgery, Southwest Hospital, Third Military Medical University,Chongqing 400038,P.R.China.
Jiang
Yaoguang
Y
Lin
Yidan
Y
Wang
Ruwen
R
Deng
Bo
B