Effect of a recombinant lentiviral vector carrying hepatocyte nuclear factor 6 gene on migration and invasion abilities of SW620 cells.
- Author:
Haitao GENG
1
;
Qian XIAO
;
Dengyong XU
;
Lifeng HU
;
Kefeng DING
Author Information
- Publication Type:Journal Article
- MeSH: Cell Line, Tumor; Cell Movement; Colorectal Neoplasms; metabolism; pathology; Genetic Vectors; genetics; Hepatocyte Nuclear Factor 6; biosynthesis; genetics; Humans; Lentivirus; genetics; metabolism; Neoplasm Invasiveness; Neoplasm Metastasis; Recombinant Proteins; biosynthesis; genetics; Transfection
- From: Journal of Southern Medical University 2012;32(1):66-70
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct a recombinant lentiviral vector that stably express hepatocyte nuclear factor 6 (HNF6) in colorectal cancer cell line and examine its effects on the invasive ability of SW620 cells.
METHODSThe lentiviral vector pLeno-DCE-HA-HNF6 was constructed and transfected into 293T cells. The supernatant containing the lentivirus particles was harvested to determine the virus titer. A stable cell line was established by infecting SW620 cells with the lentivirus particles, and the transfection efficiency was examined by fluorescence microscopy and flow cytometry. The invasion ability of the transfected SW620-HNF6 cells was assessed by wound healing and transwell assays.
RESULTSThe recombinant lentiviral vector was correctly constructed and verified by sequencing. SW620-HNF6 cell line with stable HNF6 expression was established successfully, and the transfection efficiency reached 82.3%. Western blotting and quantitative PCR demonstrated significantly upregulated HNF6 expression in SW620-HNF6 cells, which showed obviously suppressed invasive ability in wound healing and transwell assays.
CONCLUSIONWe have successfully established a colorectal cancer cell line SW620-HNF6 stably expressing HNF6, which shows a lowered migration activity in vitro.
