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Beneficial role of overexpression of TFPI-2 on tumour progression in human small cell lung cancer.

Lavergne M, Jourdan ML, Blechet C, Guyetant S, Pape AL, Heuze-Vourc'h N, Courty Y, Lerondel S, Sobilo J, Iochmann S, Reverdiau P - FEBS Open Bio (2013)

Bottom Line: In this study, low levels of TFPI-2 expression were found in 65% of patients with small cell lung cancer (SCLC), the most aggressive type of lung cancer.Such inhibition could be explained in vitro by a decrease in tumour cell viability, blockade of G1/S phase cell cycle transition and an increase in apoptosis shown in NCI-H209 cells expressing TFPI-2.We also demonstrated that TFPI-2 upregulation in NCI-H209 cells decreased MMP expression, particularly by downregulating MMP-1 and MMP-3.

View Article: PubMed Central - PubMed

Affiliation: EA 6305, Université François Rabelais de Tours, Tours F-37032, France ; Centre d'Etude des Pathologies Respiratoires, UMR 1100/EA6305, Tours F-37032, France.

ABSTRACT
Tissue factor pathway inhibitor-2 (TFPI-2) is a potent inhibitor of plasmin, a protease which is involved in tumour progression by activating (MMPs). This therefore makes TFPI-2 a potential inhibitor of invasiveness and the development of metastases. In this study, low levels of TFPI-2 expression were found in 65% of patients with small cell lung cancer (SCLC), the most aggressive type of lung cancer. To study the impact of TFPI-2 in tumour progression, TFPI-2 was overexpressed in NCI-H209 SCLC cells which were orthotopically implanted in nude mice. Investigations showed that TFPI-2 inhibited lung tumour growth. Such inhibition could be explained in vitro by a decrease in tumour cell viability, blockade of G1/S phase cell cycle transition and an increase in apoptosis shown in NCI-H209 cells expressing TFPI-2. We also demonstrated that TFPI-2 upregulation in NCI-H209 cells decreased MMP expression, particularly by downregulating MMP-1 and MMP-3. Moreover, TFPI-2 inhibited phosphorylation of the MAPK signalling pathway proteins involved in the induction of MMP transcripts, among which MMP-1 was predominant in SCLC tissues and was inversely expressed with TFPI-2 in 35% of cases. These results suggest that downregulation of TFPI-2 expression could favour the development of SCLC.

No MeSH data available.


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Impact of TFPI-2 expression on cell growth and cell cycle in NCI-H209 cells. (A) Proliferation of cells expressing TFPI-2 (T6 and T28 cells) or not (852 cells) was assessed by MTS assay every 3 days over 2 weeks. Results represent median and quartile (Q1 and Q3) from at least 4 independent experiments using Mann Whitney statistical analysis with *p < 0.05, **p < 0.01, ***p < 0.001. (B) Cell cycle phase distribution of NCI-H209 cells after serum starvation for 8 h, and exposure to 2.5 mM thymidine for 16 h and 300 μM hydroxyurea for 24 h. After culture with medium containing 10% FCS for 0, 8, 16 and 24 h, cells were stained with propidium iodide and analysed by flow cytometry. The percentages of cells in G0/G1, S, G2/M phases are expressed as the median and quartiles of at least 3 independent experiments. (C) Representative immunobloting of p15, p27 and CDK4 involved in cell cycle and β-actin as the loading control, expressed in 852, T28 and T6 cells.
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fig0004: Impact of TFPI-2 expression on cell growth and cell cycle in NCI-H209 cells. (A) Proliferation of cells expressing TFPI-2 (T6 and T28 cells) or not (852 cells) was assessed by MTS assay every 3 days over 2 weeks. Results represent median and quartile (Q1 and Q3) from at least 4 independent experiments using Mann Whitney statistical analysis with *p < 0.05, **p < 0.01, ***p < 0.001. (B) Cell cycle phase distribution of NCI-H209 cells after serum starvation for 8 h, and exposure to 2.5 mM thymidine for 16 h and 300 μM hydroxyurea for 24 h. After culture with medium containing 10% FCS for 0, 8, 16 and 24 h, cells were stained with propidium iodide and analysed by flow cytometry. The percentages of cells in G0/G1, S, G2/M phases are expressed as the median and quartiles of at least 3 independent experiments. (C) Representative immunobloting of p15, p27 and CDK4 involved in cell cycle and β-actin as the loading control, expressed in 852, T28 and T6 cells.

Mentions: To establish whether TFPI-2 could modify the cell proliferation that might influence tumour progression, a MTS assay on cells expressing TFPI-2 and on cells not expressing TFPI-2 was performed every 3 days over a period of 2 weeks. The 3 clones had the same proliferation rate for 10 days (Fig. 4A). In contrast, we clearly demonstrated that TFPI-2 expression in T6 and T28 cells significantly reduced proliferation (4-fold) from the 12th day compared to that of 852 cells.


Beneficial role of overexpression of TFPI-2 on tumour progression in human small cell lung cancer.

Lavergne M, Jourdan ML, Blechet C, Guyetant S, Pape AL, Heuze-Vourc'h N, Courty Y, Lerondel S, Sobilo J, Iochmann S, Reverdiau P - FEBS Open Bio (2013)

Impact of TFPI-2 expression on cell growth and cell cycle in NCI-H209 cells. (A) Proliferation of cells expressing TFPI-2 (T6 and T28 cells) or not (852 cells) was assessed by MTS assay every 3 days over 2 weeks. Results represent median and quartile (Q1 and Q3) from at least 4 independent experiments using Mann Whitney statistical analysis with *p < 0.05, **p < 0.01, ***p < 0.001. (B) Cell cycle phase distribution of NCI-H209 cells after serum starvation for 8 h, and exposure to 2.5 mM thymidine for 16 h and 300 μM hydroxyurea for 24 h. After culture with medium containing 10% FCS for 0, 8, 16 and 24 h, cells were stained with propidium iodide and analysed by flow cytometry. The percentages of cells in G0/G1, S, G2/M phases are expressed as the median and quartiles of at least 3 independent experiments. (C) Representative immunobloting of p15, p27 and CDK4 involved in cell cycle and β-actin as the loading control, expressed in 852, T28 and T6 cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3722576&req=5

fig0004: Impact of TFPI-2 expression on cell growth and cell cycle in NCI-H209 cells. (A) Proliferation of cells expressing TFPI-2 (T6 and T28 cells) or not (852 cells) was assessed by MTS assay every 3 days over 2 weeks. Results represent median and quartile (Q1 and Q3) from at least 4 independent experiments using Mann Whitney statistical analysis with *p < 0.05, **p < 0.01, ***p < 0.001. (B) Cell cycle phase distribution of NCI-H209 cells after serum starvation for 8 h, and exposure to 2.5 mM thymidine for 16 h and 300 μM hydroxyurea for 24 h. After culture with medium containing 10% FCS for 0, 8, 16 and 24 h, cells were stained with propidium iodide and analysed by flow cytometry. The percentages of cells in G0/G1, S, G2/M phases are expressed as the median and quartiles of at least 3 independent experiments. (C) Representative immunobloting of p15, p27 and CDK4 involved in cell cycle and β-actin as the loading control, expressed in 852, T28 and T6 cells.
Mentions: To establish whether TFPI-2 could modify the cell proliferation that might influence tumour progression, a MTS assay on cells expressing TFPI-2 and on cells not expressing TFPI-2 was performed every 3 days over a period of 2 weeks. The 3 clones had the same proliferation rate for 10 days (Fig. 4A). In contrast, we clearly demonstrated that TFPI-2 expression in T6 and T28 cells significantly reduced proliferation (4-fold) from the 12th day compared to that of 852 cells.

Bottom Line: In this study, low levels of TFPI-2 expression were found in 65% of patients with small cell lung cancer (SCLC), the most aggressive type of lung cancer.Such inhibition could be explained in vitro by a decrease in tumour cell viability, blockade of G1/S phase cell cycle transition and an increase in apoptosis shown in NCI-H209 cells expressing TFPI-2.We also demonstrated that TFPI-2 upregulation in NCI-H209 cells decreased MMP expression, particularly by downregulating MMP-1 and MMP-3.

View Article: PubMed Central - PubMed

Affiliation: EA 6305, Université François Rabelais de Tours, Tours F-37032, France ; Centre d'Etude des Pathologies Respiratoires, UMR 1100/EA6305, Tours F-37032, France.

ABSTRACT
Tissue factor pathway inhibitor-2 (TFPI-2) is a potent inhibitor of plasmin, a protease which is involved in tumour progression by activating (MMPs). This therefore makes TFPI-2 a potential inhibitor of invasiveness and the development of metastases. In this study, low levels of TFPI-2 expression were found in 65% of patients with small cell lung cancer (SCLC), the most aggressive type of lung cancer. To study the impact of TFPI-2 in tumour progression, TFPI-2 was overexpressed in NCI-H209 SCLC cells which were orthotopically implanted in nude mice. Investigations showed that TFPI-2 inhibited lung tumour growth. Such inhibition could be explained in vitro by a decrease in tumour cell viability, blockade of G1/S phase cell cycle transition and an increase in apoptosis shown in NCI-H209 cells expressing TFPI-2. We also demonstrated that TFPI-2 upregulation in NCI-H209 cells decreased MMP expression, particularly by downregulating MMP-1 and MMP-3. Moreover, TFPI-2 inhibited phosphorylation of the MAPK signalling pathway proteins involved in the induction of MMP transcripts, among which MMP-1 was predominant in SCLC tissues and was inversely expressed with TFPI-2 in 35% of cases. These results suggest that downregulation of TFPI-2 expression could favour the development of SCLC.

No MeSH data available.


Related in: MedlinePlus