Construction and identification of an antisense glucose transporter-1 plasmid.
Our aim was to construct a pcDNA3.1(+) eucaryotic expression system vector containing the antisense glucose transporter-1 (Glut-1) gene. Total RNA was isolated from human Hep-2 laryngeal carcinoma cells, and the Glut-1 and antisense Glut-1 sequences were amplified by polymerase chain reaction. Expression plasmids containing the sense and antisense cDNA were constructed using the pcDNA3.1(+) vector. The resulting sense and antisense vectors, pcDNA3.1(+)-Glut-1 and pcDNA3.1(+)-antiGlut-1, respectively, were examined by restriction analysis and DNA sequencing. The pcDNA3.1(+)-antiGlut-1 was subsequently transfected into Hep-2 cells. AntiGlut-1 mRNA expression was detected, indicating the successful construction of an antisense Glut-1 plasmid capable of transfecting Hep-2 laryngeal carcinoma cells. These data provide a firm basis for additional studies using the plasmid pcDNA3.1(+)-antiGlut-1 to determine its therapeutic potential for the treatment of laryngeal carcinoma.
Chen
X M
XM
Key Laboratory of National Infectious Diseases, Institute of Infectious Diseases, Zhejiang University, Hangzhou, China.
Zhou
S H
SH
Fan
J
J
Hu
M J
MJ
Wang
S Q
SQ
Yu
Y S
YS
D000818:Animals; D001483:Base Sequence; D045744:Cell Line, Tumor; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D051272:Glucose Transporter Type 1 / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007822:Laryngeal Neoplasms / Q000235:genetics / Q000628:therapy; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism
D051272:Glucose Transporter Type 1; D016376:Oligonucleotides, Antisense