[Study in vitro on p16beta interfering the cell cycle signal conduction of laryngeal squamous cell carcinoma].
OBJECTIVE
To investigate the effect of p16beta interfering with the signal conduction of Hep-2 cell cycle in Vitro.


METHODS
AdEasy Vector System was used to construct the recombinant adenovirus vector AdEasy-GFP-p1613. The recombinant adenovirus vector could introduce p16beta gene into HEK 293 cell. Then the purified recombinant adenovirus was used to infect Hep-2 cells in vitro. The protein expression of p16beta, proliferation inhibition, cell cycle arrest, DNA content, apoptosis ratio in Hep-2 cells were examined by MTT, Western blotting analysis, Flow cytometry assay, Immunocytochemistry respectively.


RESULTS
The recombinant adenovirus Adeasy-p16beta was constructed successfully and higher titer recombinant adenovirus particle was got. When Adeasy-p16beta was transferred into Hep-2 cells, both the growth inhibition and P14 (ARF) protein overexpression in Hep-2 cells were visible effectively. The most Hep-2 cells were blocked in G1/G0 phase and the cell apoptosis ratio increased.


CONCLUSIONS
The p16beta in Hep-2 cell infected by recombinant adenovirus, which lead to express P14 (ARF) protein effectively and to inhibit cell proliferative activation, effectively interfere the signal conduction mechanisms of culture Hep-2 laryngeal squamous cell carcinoma, and the growth of much cultured cells is blocked in G1/G0 phase.
Fan
Min
M
Department of Otolaryngology, West China Hospital, Sichuan University, Chengdu 610041, China.
Xian
Jun-ming
JM
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; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000096:biosynthesis* / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007822:Laryngeal Neoplasms / Q000473:pathology*; D010957:Plasmids / Q000235:genetics; D011995:Recombination, Genetic; D015398:Signal Transduction; D014162:Transfection; D014407:Tumor Cells, Cultured
D019941:Cyclin-Dependent Kinase Inhibitor p16