Establishment of a lung adenocarcinoma cell line stably expressing small interfering RNA targeting hepatoma-derived growth factor and detection of interference effect / 南方医科大学学报

ABSTRACT

<p><b>OBJECTIVE</b>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.</p><p><b>METHOD</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>