ZHANG Qian-ying,YE Lin,YU Xiao-yan,SHEN Na,DENG Bi,WU Xiao-song,CHEN Hong-yan.Effect of focal adhesion kinase gene silencing on proliferation and invasion of Hep-2 cells[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(06):526-529.
ZHANG Qian-ying,YE Lin,YU Xiao-yan,SHEN Na,DENG Bi,WU Xiao-song,CHEN Hong-yan.Effect of focal adhesion kinase gene silencing on proliferation and invasion of Hep-2 cells[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(06):526-529.DOI:
Effect of focal adhesion kinase gene silencing on proliferation and invasion of Hep-2 cells
Objective To observe the effects of RNAi-mediated focal adhesion kinase (FAK) gene silencing on laryngeal carcinoma Hep-2 cells. Methods Recombinant plasmids generating short hairpin RNA were constructed
including two interferential sequences FAK-pGenesil-CH1
FAK-pGenesil-CH2 and one negative control sequence FAK-pGenesil-HK. Hep-2 cells were transfected with recombinant plasmids. FAK mRNA and protein were
analyzed respectively by RT-PCR and Western blotting. Cell proliferation
cell cycle and invasion of Hep-2 cells were observed by MTT assay
flow cytometry and transwell invasion assay
respectively. Results Recombinant plasmids of FAK-pGenesil-CH1 and FAK-pGenesil-CH2 downregulated the expressions of FAK mRNA and protein (P0.05). The proliferation inhibition ratio in Hep-2 cells reached 58.34% and 52.78% 48 h after transfection of recombinant plasmids of FAK-pGenesil-CH1 and FAK-pGenesil-CH2
while that was 8.34% in the control (P0.05). More cells arrested in G
0
/G
1
phase in the transfected groups than controls. The number of FAK-pGenesil-CH1 transfected Hep-2 cells
and FAK-pGenesil-CH2 transfected Hep-2 cells through Matrigel membrane was (56.0±6.7) and (51.0±5.1)
respectively
significantly less than that in control groups (P0.05). Conclusion The constructed recombinant plasmids of FAK-pGenesil-CH1 and FAK-pGenesil-CH2 can suppress the expression of FAK mRNA and protein in Hep-2 cells
and inhibit the cloning and invasion of Hep-2 cells.