[Adenovirus-mediated antisense HSP70 cDNA transfection inhibits the growth of laryngeal carcinoma Hep-2 cells].
OBJECTIVE
To construct a recombinant adenovirus vector carrying antisense heat shock protein 70 (HSP70) cDNA and observe its effect on inhibiting the growth of laryngeal carcinoma Hep-2 cells.


METHODS
The HSP70 gene fragment encoding the 5' region was cloned reversely into the shuttle plasmid PAdTrack-CMV, and the resultant plasmid was recombined with the backbone plasmid PadEasy-1 in the E.coli Bj5183 cells to generate the recombinant adenovirus vector. The adenovirus were then packaged and amplified in 293 cells, and the viral titer was determined using GFP.


RESULTS
The recombinant adenovirus vector carrying antisense HSP70 cDNA was constructed successfully with a viral titer of 8 x 10(9). HSP70 expression of Hep-2 cells was obviously blocked by antisense HSP70 RNA, and Western blotting and immuohistochemistry demonstrated that cells transfected with antisense HSP70 did not express or express HSP70 at low levels. Flow cytometry presented apoptotic peak in the antisense HSP70-transfected cells, but not in the control cells.


CONCLUSIONS
The recombinant adenovirus vector containing antisense HSP70 cDNA can effectively deliver antisense HSP70 gene into Hep-2 cells, suggesting the great potential of this gene therapy strategy in management of human laryngeal carcinoma.
Wang
Xiao-xia
XX
Department of Otolaryngology, 451 Hospital of PLA, Xi'an 710054, China. xiaoxiawang9139@sina.com.cn
Yao
Xiao-bao
XB
Ji
Xian-sheng
XS
Li
Sheng-li
SL
Zhu
Hong-liang
HL
Fan
Dai-ming
DM
D000256:Adenoviridae / Q000235:genetics; D045744:Cell Line, Tumor; D016373:DNA, Antisense / Q000494:pharmacology*; D018076:DNA, Complementary / Q000235:genetics; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000096:biosynthesis; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics*; D006801:Humans; D007822:Laryngeal Neoplasms / Q000628:therapy*; D016372:RNA, Antisense / Q000494:pharmacology; D014162:Transfection
D016373:DNA, Antisense; D018076:DNA, Complementary; D018840:HSP70 Heat-Shock Proteins; D016372:RNA, Antisense