Construction of human Wnt-5a sense gene and RNAi eukaryotic expression vector.
OBJECTIVE
Wnt-5a is important in the physiological development and differentiation of lung, and is also involved in the regulating proliferation, differentiation and invasion of tumor cells. However, very little is known about the roles of Wnt-5a in the development of lung cancer. The porpus of this study was to explore the role of Wnt-5a in the development of the non-small cell lung cancer, through constructing plasmids containing Wnt-5a sense gene siRNA.


METHODS
We constructed the plasmids containing the Wnt-5a sense gene and siRNA eukaryotic expression vector and transfected it into the human lung squamous carcinoma cell line H157 and adenocarcinoma cell line A549. Expressions of Wnt-5a RNA and protein and proliferation of the cells were measured.


RESULTS
Expression of Wnt-5a protein significantly stimulated cell proliferation, and transfection with siRNA plasmids suppressed Wnt-5 expression and cell proliferation.


CONCLUSIONS
The human plasmids containing Wnt-5a sense gene and siRNA eukaryotic expression vector was successfully constructed, and transfection to human cancer cells induces cell proliferation. siRNA actively suppressed Wnt-5a expression.
Huang
Ying
Y
Department of Respiratory Medicine, The Third Military Medical University, Chongqing, China.
Xiong
Wei
W
Liu
Guoxiang
G
Zhang
Bo
B
Xu
Gang
G
Yang
Heping
H
D001483:Base Sequence; D015153:Blotting, Western; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D003062:Codon / Q000235:genetics*; D004247:DNA / Q000096:biosynthesis / Q000235:genetics; D005057:Eukaryotic Cells; D005819:Genetic Markers; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D010957:Plasmids / Q000235:genetics; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D034741:RNA, Small Interfering / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051153:Wnt Proteins / Q000235:genetics*; D000071818:Wnt-5a Protein
D003062:Codon; D005819:Genetic Markers; D011518:Proto-Oncogene Proteins; D034741:RNA, Small Interfering; C085389:WNT5A protein, human; D051153:Wnt Proteins; D000071818:Wnt-5a Protein; D004247:DNA