Establishment of a lung adenocarcinoma cell line stably expressing small interfering RNA targeting hepatoma-derived growth factor and detection of interference effect.
- Author:
Jie MENG
1
,
2
;
Li-Ming CAO
;
Cheng-Ping HU
;
Zhi-Yuan ZHEN
Author Information
- Publication Type:Journal Article
- MeSH: Adenocarcinoma; metabolism; pathology; Base Sequence; Cell Line, Tumor; Female; Genetic Vectors; genetics; Humans; Intercellular Signaling Peptides and Proteins; biosynthesis; genetics; Lentivirus; genetics; metabolism; Lung Neoplasms; metabolism; pathology; Male; Molecular Sequence Data; RNA Interference; RNA, Small Interfering; genetics; Recombinant Proteins; biosynthesis; genetics; Sequence Analysis, DNA
- From: Journal of Southern Medical University 2009;29(11):2233-2236
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct a small interfering RNA (siRNA) expression vector targeting hepatoma-derived growth factor (HDGF) and establish a lung adenocarcinoma cell line stably expressing siRNA-HDGF.
METHODRT-PCR was used to examine HDGF expression in lung adenocarcinoma samples and the matched adjacent lung tissues, and also in lung adenocarcinoma SPC-A-1 cell line. A recombinant lentivirus shRNA-HDGF vector was constructed and transfected into SPC-A-1 cells via Lipofectamine 2000, and the cells with stable expression of HDGF-siRNA was screened by blasticidin selection. The interference effect of siRNA-HDGF was assessed by real-time PCR.
RESULTSCompared to the adjacent lung tissues, lung adenocarcinoma and SPC-A-1 cells showed increased expression of HDGF. The recombinant lentivirus shRNA-HDGF vector was successfully constructed and verified by sequence analysis. siRNA-HDGF recombinants markedly inhibited the expression of HDGF in SPC-A-1 cells.
CONCLUSIONHDGF expression increases in lung adenocarcinoma and SPC-A-1 cell lines. The recombinant siRNA-HDGF lentivirus vector can inhibit the expression of HDGF in SPC-A-1 cells.
