[Construction of the recombinant adenovirus vector with EGFR antisense cDNA of controlling cell cycle].
OBJECTIVE
To recombine the adenovirus vector carrying EGFR sence/antisense cDNA which takes part in control of cell cycle.


METHODS
The 1032 bp EGFR sence/antisense cDNA fragment was cloned into the shuttle plasmid pAdTrack-CMV. The resultant plasmid and the backbone plasmid pAdEasy-1 were transferred into E. coli BJ5183 for homologous recombination, and the recombinant adenoviruses were generated in cells. The recombinant adenoviruses were packaged and amplified in the 293 cells. Then the viral titer was detected by GFP.


RESULTS
The recombinant adenovirus vector carrying EGFR sence/antisense cDNA to control the cell cycle was constructed successfully. The viral titers were 2.2 x 10(9) efu/mL and 2.5 X 10(9) efu/mL respectively.


CONCLUSIONS
The recombinant adenovirus vector constructed by us could introduce EGFR antisense cDNA into the laryngeal squamous cell carcinoma line or tumor tissue, which would provide an experiment basis to study further the interfered mechanism of signal transdution and the therapies of laryngeal squamous cell carcinoma.
Xian
Jun-Ming
JM
Department of Otolaryngology, West China Hospital, Sichuan University, Chengdu 610041, China.
Zhou
Guang-yao
GY
Liang
Chuan-yu
CY
Liu
Shi-xi
SX
D000256:Adenoviridae / Q000235:genetics* / Q000378:metabolism; D002294:Carcinoma, Squamous Cell / Q000473:pathology; D002453:Cell Cycle; D018076:DNA, Complementary / Q000235:genetics; D004622:Embryo, Mammalian; D066246:ErbB Receptors / Q000096:biosynthesis* / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D007668:Kidney / Q000166:cytology; D007822:Laryngeal Neoplasms / Q000473:pathology; D016376:Oligonucleotides, Antisense / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D011995:Recombination, Genetic; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; D016376:Oligonucleotides, Antisense; D011994:Recombinant Proteins; D066246:ErbB Receptors