[Establishment of a lung adenocarcinoma cell line stably expressing small interfering RNA targeting hepatoma-derived growth factor and detection of interference effect].
OBJECTIVE
To 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.


METHODS
RT-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.


RESULTS
Compared 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.


CONCLUSIONS
HDGF 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.
Meng
Jie
J
Department of Respiration, Xiangya Hospital, Central South University, Changsha 410008, China. xw7018@163.com
Cao
Li-Ming
LM
Hu
Cheng-Ping
CP
Zhen
Zhi-Yuan
ZY
D000230:Adenocarcinoma / Q000378:metabolism* / Q000473:pathology; D001483:Base Sequence; D045744:Cell Line, Tumor; D005260:Female; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000096:biosynthesis* / Q000235:genetics; D016086:Lentivirus / Q000235:genetics / Q000378:metabolism; D008175:Lung Neoplasms / Q000378:metabolism* / Q000473:pathology; D008297:Male; D008969:Molecular Sequence Data; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D017422:Sequence Analysis, DNA
D036341:Intercellular Signaling Peptides and Proteins; D034741:RNA, Small Interfering; D011994:Recombinant Proteins; C051610:hepatoma-derived growth factor