Effect of lentivirus-mediated nestin gene silencing on metastatic potential of human melanoma cell line UACC903 in vitro.
- Author:
Xuhui YANG
1
;
Tian XIA
;
Weihua YU
;
Xiaofang LU
;
Peng XIANG
;
Feng HE
Author Information
- Publication Type:Journal Article
- MeSH: Cell Line, Tumor; Cell Movement; genetics; Genetic Vectors; Humans; Intermediate Filament Proteins; genetics; metabolism; Lentivirus; genetics; Melanoma; pathology; Neoplasm Metastasis; genetics; Nerve Tissue Proteins; genetics; metabolism; Nestin; RNA Interference; RNA, Messenger; genetics; metabolism; RNA, Small Interfering; genetics
- From: Journal of Southern Medical University 2012;32(1):7-13
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the effect of lentivirus-mediated RNA interference (RNAi) of nestin on the metastatic potential of human melanoma cell line UACC903.
METHODSA lentiviral vector for RNAi targeting the coding region of human nestin mRNA (nestin-RNAi-LV) and another control vector containing a nonsense sequence were constructed. The vectors were transfected into UACC903 cells, and nestin expression in the cells was detected by RT-PCR, immunofluorence assay and Western blotting. The invasive ability and migration of the transfected UACC903 cells was evaluated using Transwell and scrape assay, respectively. Fluorescence assay was used to examine the expressions of E-cadherin, N-cadherin and β-catenin in the cells.
RESULTSThe lentiviral vector nestin-RNAi-LV was constructed successfully. Compared with the control vector, nestin-RNAi-LV resulted in obviously reduced expression of nestin mRNA and protein, lowered migration ability of UACC903 cells, and reduced cell adhesion and invasiveness (P<0.05).
CONCLUSIONLentivirus-mediated nestin RNAi can specifically inhibit nestin expression to cause decreased cell migration and invasiveness of human melanoma cell line UACC903.
