Fan Min, Xian Jun-ming, Zhou Guang-yao, Liang Chuan-yu, Liu Shi-xi
Department of Otolaryngology, West China Hospital, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2006 Jul;37(4):515-9.
To investigate the effect of p16beta interfering with the signal conduction of Hep-2 cell cycle in Vitro.
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.
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.
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.
