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Small interference RNA targeting tissue factor inhibits human lung adenocarcinoma growth in vitro and in vivo.

Xu C, Gui Q, Chen W, Wu L, Sun W, Zhang N, Xu Q, Wang J, Fu X - J. Exp. Clin. Cancer Res. (2011)

Bottom Line: TF -siRNA significantly reduced the expression of TF in the mRNA and protein levels.The down-regulation of TF in A549 cells resulted in the suppression of cell proliferation, invasion and metastasis and induced cell apoptosis in dose-dependent manner.Erk MAPK, PI3K/Akt pathways as well as VEGF and MMP-2/-9 expressions were inhibited in TF-siRNA transfected cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of General Thoracic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People's Republic of China.

ABSTRACT

Background: The human coagulation trigger tissue factor (TF) is overexpressed in several types of cancer and involved in tumor growth, vascularization, and metastasis. To explore the role of TF in biological processes of lung adenocarcinoma, we used RNA interference (RNAi) technology to silence TF in a lung adenocarcinoma cell line A549 with high-level expression of TF and evaluate its antitumor effects in vitro and in vivo.

Methods: The specific small interfering RNA (siRNA) designed for targeting human TF was transfected into A549 cells. The expression of TF was detected by reverse transcription-PCR and Western blot. Cell proliferation was measured by MTT and clonogenic assays. Cell apoptosis was assessed by flow cytometry. The metastatic potential of A549 cells was determined by wound healing, the mobility and Matrigel invasion assays. Expressions of PI3K/Akt, Erk1/2, VEGF and MMP-2/-9 in transfected cells were detected by Western blot. In vivo, the effect of TF-siRNA on the growth of A549 lung adenocarcinoma xenografts in nude mice was investigated.

Results: TF -siRNA significantly reduced the expression of TF in the mRNA and protein levels. The down-regulation of TF in A549 cells resulted in the suppression of cell proliferation, invasion and metastasis and induced cell apoptosis in dose-dependent manner. Erk MAPK, PI3K/Akt pathways as well as VEGF and MMP-2/-9 expressions were inhibited in TF-siRNA transfected cells. Moreover, intratumoral injection of siRNA targeting TF suppressed the tumor growth of A549 cells in vivo model of lung adenocarcinoma.

Conclusions: Down-regulation of TF using siRNA could provide a potential approach for gene therapy against lung adenocarcinoma, and the antitumor effects may be associated with inhibition of Erk MAPK, PI3K/Akt pathways.

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Related in: MedlinePlus

Knockdown of TF with TF-siRNA induced apoptosis of lung adenocarcinoma cells. The transfected cells, labeled with AnnexinV-FITC and propidium iodide, were subjected to flow cytometric analysis. Two parameter histogram Dot Plot displayed FL1-FITC on the x axis and FL2-PI on the y axis. The result showed that TF-siRNA increased the apoptotic rate in A549 cells in a dose-dependent manner.
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Figure 13: Knockdown of TF with TF-siRNA induced apoptosis of lung adenocarcinoma cells. The transfected cells, labeled with AnnexinV-FITC and propidium iodide, were subjected to flow cytometric analysis. Two parameter histogram Dot Plot displayed FL1-FITC on the x axis and FL2-PI on the y axis. The result showed that TF-siRNA increased the apoptotic rate in A549 cells in a dose-dependent manner.

Mentions: To evaluate further whether knockdown of TF induces A549 cells apoptosis, at 48 h after transfection, the cells were harvested and analyzed by flow cytometry. As shown in Figure 13, the apoptosis rates of 25 nM, 50 nM and 100 nM SiTF groups were 7.0%, 9.0% and 16.0%, respectively, which were higher than 4.0% in control and 4.8% in mock groups, and indicated a dose-dependent increase.


Small interference RNA targeting tissue factor inhibits human lung adenocarcinoma growth in vitro and in vivo.

Xu C, Gui Q, Chen W, Wu L, Sun W, Zhang N, Xu Q, Wang J, Fu X - J. Exp. Clin. Cancer Res. (2011)

Knockdown of TF with TF-siRNA induced apoptosis of lung adenocarcinoma cells. The transfected cells, labeled with AnnexinV-FITC and propidium iodide, were subjected to flow cytometric analysis. Two parameter histogram Dot Plot displayed FL1-FITC on the x axis and FL2-PI on the y axis. The result showed that TF-siRNA increased the apoptotic rate in A549 cells in a dose-dependent manner.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3118969&req=5

Figure 13: Knockdown of TF with TF-siRNA induced apoptosis of lung adenocarcinoma cells. The transfected cells, labeled with AnnexinV-FITC and propidium iodide, were subjected to flow cytometric analysis. Two parameter histogram Dot Plot displayed FL1-FITC on the x axis and FL2-PI on the y axis. The result showed that TF-siRNA increased the apoptotic rate in A549 cells in a dose-dependent manner.
Mentions: To evaluate further whether knockdown of TF induces A549 cells apoptosis, at 48 h after transfection, the cells were harvested and analyzed by flow cytometry. As shown in Figure 13, the apoptosis rates of 25 nM, 50 nM and 100 nM SiTF groups were 7.0%, 9.0% and 16.0%, respectively, which were higher than 4.0% in control and 4.8% in mock groups, and indicated a dose-dependent increase.

Bottom Line: TF -siRNA significantly reduced the expression of TF in the mRNA and protein levels.The down-regulation of TF in A549 cells resulted in the suppression of cell proliferation, invasion and metastasis and induced cell apoptosis in dose-dependent manner.Erk MAPK, PI3K/Akt pathways as well as VEGF and MMP-2/-9 expressions were inhibited in TF-siRNA transfected cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of General Thoracic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People's Republic of China.

ABSTRACT

Background: The human coagulation trigger tissue factor (TF) is overexpressed in several types of cancer and involved in tumor growth, vascularization, and metastasis. To explore the role of TF in biological processes of lung adenocarcinoma, we used RNA interference (RNAi) technology to silence TF in a lung adenocarcinoma cell line A549 with high-level expression of TF and evaluate its antitumor effects in vitro and in vivo.

Methods: The specific small interfering RNA (siRNA) designed for targeting human TF was transfected into A549 cells. The expression of TF was detected by reverse transcription-PCR and Western blot. Cell proliferation was measured by MTT and clonogenic assays. Cell apoptosis was assessed by flow cytometry. The metastatic potential of A549 cells was determined by wound healing, the mobility and Matrigel invasion assays. Expressions of PI3K/Akt, Erk1/2, VEGF and MMP-2/-9 in transfected cells were detected by Western blot. In vivo, the effect of TF-siRNA on the growth of A549 lung adenocarcinoma xenografts in nude mice was investigated.

Results: TF -siRNA significantly reduced the expression of TF in the mRNA and protein levels. The down-regulation of TF in A549 cells resulted in the suppression of cell proliferation, invasion and metastasis and induced cell apoptosis in dose-dependent manner. Erk MAPK, PI3K/Akt pathways as well as VEGF and MMP-2/-9 expressions were inhibited in TF-siRNA transfected cells. Moreover, intratumoral injection of siRNA targeting TF suppressed the tumor growth of A549 cells in vivo model of lung adenocarcinoma.

Conclusions: Down-regulation of TF using siRNA could provide a potential approach for gene therapy against lung adenocarcinoma, and the antitumor effects may be associated with inhibition of Erk MAPK, PI3K/Akt pathways.

Show MeSH
Related in: MedlinePlus