Soluble FasL combined with survivin RNA interference induced apoptosis of lung adenocarcinoma
- Author:
Li-Ming LIU
1
Author Information
1. Department of Respiratory Medicine
- Publication Type:Journal Article
- Keywords:
Apoptosis;
Cell line, tumor;
Fas ligand protein;
RNA interference
- From:
Tumor
2007;27(8):615-619
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the anti-proliferative and pro-apoptotic effects of soluble Fas ligand (sFasL) combined with survivin RNA interference on A549 lung adenocarcinoma cells in vitro. Methods: The expression vector of survivin small interfering RNA (siRNA) was constructed and confirmed by sequencing. The survivin siRNA expression vector was transfected into A549 cells and the stable cell line expressed survivin small interfering RNA were selected by G418 (geneticin) screening. The mRNA and protein levels of survivin were tested by fluorescent quantitative real-time reverse transcription polymerase chain reaction (FQ-RT-PCR), Western blot, and immunohistochemistry. After the transfected cells were treated with sFasL, the cell proliferation and apoptosis were assessed by MTT assay and flow cytometry. Results: Survivin siRNA expression vector was successfully constructed and stable A549 cells were selected. The mRNA and protein levels of survivin decreased by 76.4% (P <0.01) and 84.5%, respectively. Immunohistochemistry showed that the survivin was down-regulated. Cells in siRNA + sFasL group had lower proliferation rate at every time point than that of single sFasL group, siRNA group and control group. The difference was significant (P < 0.01). The apoptotic rate was significantly higher in siRNA + sFasL group than that of sFasL group, siRNA group, and control group [(18.6 ± 2.93)% vs (10.12 ± 2.31)%, (10.09 ± 1.31)%, (1.14 ± 0.21)%, P < 0.01]. Conclusions: Survivin siRNA expression vector significantly inhibites the expression of survivin in A549 cells. The sFasL combined with survivin siRNA interference can more effectively inhibit the proliferation and induce the apoptosis than single sFasL or survivin RNA interference treatment in lung adeno carcinoma A549 cells.
