Construction of lentiviral vector of RNA inter-ference of MMP-9 gene and its inhibitive role on the invasion of laryngeal cancer cell
- VernacularTitle:MMP-9基因RNAi重组慢病毒的构建及其对喉癌细胞侵袭的抑制
- Author:
Yanan SUN
;
Ming LIU
;
Yan SUN
;
Linli TIAN
;
Hui JIAO
;
- Publication Type:Journal Article
- Keywords:
RNA Interference;
Gelatinase B;
Neoplasms;
Lentivirus
- From:
Chinese Archives of Otolaryngology-Head and Neck Surgery
2006;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE To construct a lentiviral vector of RNA interference (RNAi) of MMP-9 gene and observe its inhibitive role on the invasion of laryngeal cancer cells.METHODS The effective sequence of siRNA targeting MMP-9 gene was confirmed.Both sense and antisense Oligo DNA of the targeting sequence was designed,synthesized and cloned into the pLVTHM vector,which contained H1 promoter and green fluorescent protein (GFP).The resulting lentiviral vector containing MMP-9 shRNA was called LV2 shMMP-9,and it was confirmed by PCR and sequencing.After that,MMP-9 shRNA was transfected into Hep-2 cells and western blot was used to test the expression of MMP-9.At last,Boyden Chamber was used to observe the invasion of the Hep-2 cells. RESULTS PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of MMP-9 (LV2shMMP-9) producing MMP-9 shRNA was constructed successfully. The titer of concentrated virus was 8?1010TU /L. Western blot showed that the expression of MMP-9 was negative in the MMP-9 siRNA transfected Hep-2 cells. And Boyden Chamber showed the invasive capability of Hep-2 cells transfected MMP-9 siRNA were obviously decreased.CONCLUSION The lentivirus RNAi vector of MMP-9 was constructed successfully,and MMP-9 silence can inhibit invasion of laryngeal cancer in vitro.
