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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.


Related in: MedlinePlus

Immunohistochemistry analysis of small cell lung cancer samples (SCLC). (A) Immunostaining of neuroendocrine and epithelial differentiation markers, chromogranin, synaptophysin, CD56 and KL1 on tissue sections from patients with SCLC. (B) Immunostaining of TFPI-2 using labelled streptavidin–biotin immunoenzymatic detection after incubation with a rabbit polyclonal antibody specific to human TFPI-2 and haematoxylin counterstaining. Representative samples with immunostaining intensity from 0 to 3. Original magnification ×40. (C) Immunostaining intensity of MMP-1, -2, -3 and -9 according to low (intensity 0 and 1) or high level of TFPI-2 expression (intensity 2 and 3) for each of the 40 tissue samples.
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fig0001: Immunohistochemistry analysis of small cell lung cancer samples (SCLC). (A) Immunostaining of neuroendocrine and epithelial differentiation markers, chromogranin, synaptophysin, CD56 and KL1 on tissue sections from patients with SCLC. (B) Immunostaining of TFPI-2 using labelled streptavidin–biotin immunoenzymatic detection after incubation with a rabbit polyclonal antibody specific to human TFPI-2 and haematoxylin counterstaining. Representative samples with immunostaining intensity from 0 to 3. Original magnification ×40. (C) Immunostaining intensity of MMP-1, -2, -3 and -9 according to low (intensity 0 and 1) or high level of TFPI-2 expression (intensity 2 and 3) for each of the 40 tissue samples.

Mentions: Biopsy specimens of SCLC obtained from 40 patients (30 men and 10 women) aged 45–84 years (median 65 years) were studied. Ninety-seven percent of these patients were smokers. Twenty-five patients (63%) had advanced disease with metastases at the time of diagnosis. Only 2 of 36 patients for whom survival data were available were still alive at the last follow up. The median overall survival was 6 months and the 2-year survival rate was 5.5%. The diagnosis of SCLC was made primarily on haematoxylin–eosin–safran stained slides showing small malignant cells with high mitotic rates. Among the epithelial markers (i.e. pan-cytokeratins (KL1) and epithelial membrane antigen (EMA)) and neuroendocrine differentiation markers (i.e. CD56, chromogranin A and synaptophysin) at least three of them were expressed in all tumours (Fig. 1A). We then examined TFPI-2 and MMP-1, -2, -3 and -9 expression in lung samples of 40 patients with SCLC and established staining intensity from 0 to 3 by immunohistochemistry. Homogeneous expression of TFPI-2 and MMP was observed in 100% of tumour cells in lung samples. Low levels of TFPI-2 (staining intensity of 0 or 1) were found in 26 (65%) SCLC specimens while high levels of TFPI-2 expression were found in only 14 (35%) SCLC specimens, including only 1 with a staining intensity of 3 (Fig. 1B). The highest expression level in MMPs was found for MMP-1 with a staining intensity of 2 for 50% of specimens (Fig. 1C). For MMP-2, MMP-3 and MMP-9, an immunostaining intensity of 1 was predominant in 50%, 57.5% and 61.5% of biopsies, respectively. To determine the relationship between TFPI-2 and MMPs, we compared the immunostaining intensity obtained from the same tissue. In SCLC with low levels of TFPI-2 (0 and 1), MMP staining was mainly equal to that of TFPI-2 or was higher for MMP-1 in only 38% of tissue samples (intensity of 2). In tissues with high levels of TFPI-2 (2 and 3), none had higher levels of MMP staining than those for TFPI-2. Staining levels for MMP-1 were mostly equal to those for TFPI-2, and staining levels lower than those for TFPI-2 were obtained for MMP-2 in all specimens, whereas for MMP-3 and MMP-9 they were lower in 85.7% and 57.1% of tissues, respectively.


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)

Immunohistochemistry analysis of small cell lung cancer samples (SCLC). (A) Immunostaining of neuroendocrine and epithelial differentiation markers, chromogranin, synaptophysin, CD56 and KL1 on tissue sections from patients with SCLC. (B) Immunostaining of TFPI-2 using labelled streptavidin–biotin immunoenzymatic detection after incubation with a rabbit polyclonal antibody specific to human TFPI-2 and haematoxylin counterstaining. Representative samples with immunostaining intensity from 0 to 3. Original magnification ×40. (C) Immunostaining intensity of MMP-1, -2, -3 and -9 according to low (intensity 0 and 1) or high level of TFPI-2 expression (intensity 2 and 3) for each of the 40 tissue samples.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig0001: Immunohistochemistry analysis of small cell lung cancer samples (SCLC). (A) Immunostaining of neuroendocrine and epithelial differentiation markers, chromogranin, synaptophysin, CD56 and KL1 on tissue sections from patients with SCLC. (B) Immunostaining of TFPI-2 using labelled streptavidin–biotin immunoenzymatic detection after incubation with a rabbit polyclonal antibody specific to human TFPI-2 and haematoxylin counterstaining. Representative samples with immunostaining intensity from 0 to 3. Original magnification ×40. (C) Immunostaining intensity of MMP-1, -2, -3 and -9 according to low (intensity 0 and 1) or high level of TFPI-2 expression (intensity 2 and 3) for each of the 40 tissue samples.
Mentions: Biopsy specimens of SCLC obtained from 40 patients (30 men and 10 women) aged 45–84 years (median 65 years) were studied. Ninety-seven percent of these patients were smokers. Twenty-five patients (63%) had advanced disease with metastases at the time of diagnosis. Only 2 of 36 patients for whom survival data were available were still alive at the last follow up. The median overall survival was 6 months and the 2-year survival rate was 5.5%. The diagnosis of SCLC was made primarily on haematoxylin–eosin–safran stained slides showing small malignant cells with high mitotic rates. Among the epithelial markers (i.e. pan-cytokeratins (KL1) and epithelial membrane antigen (EMA)) and neuroendocrine differentiation markers (i.e. CD56, chromogranin A and synaptophysin) at least three of them were expressed in all tumours (Fig. 1A). We then examined TFPI-2 and MMP-1, -2, -3 and -9 expression in lung samples of 40 patients with SCLC and established staining intensity from 0 to 3 by immunohistochemistry. Homogeneous expression of TFPI-2 and MMP was observed in 100% of tumour cells in lung samples. Low levels of TFPI-2 (staining intensity of 0 or 1) were found in 26 (65%) SCLC specimens while high levels of TFPI-2 expression were found in only 14 (35%) SCLC specimens, including only 1 with a staining intensity of 3 (Fig. 1B). The highest expression level in MMPs was found for MMP-1 with a staining intensity of 2 for 50% of specimens (Fig. 1C). For MMP-2, MMP-3 and MMP-9, an immunostaining intensity of 1 was predominant in 50%, 57.5% and 61.5% of biopsies, respectively. To determine the relationship between TFPI-2 and MMPs, we compared the immunostaining intensity obtained from the same tissue. In SCLC with low levels of TFPI-2 (0 and 1), MMP staining was mainly equal to that of TFPI-2 or was higher for MMP-1 in only 38% of tissue samples (intensity of 2). In tissues with high levels of TFPI-2 (2 and 3), none had higher levels of MMP staining than those for TFPI-2. Staining levels for MMP-1 were mostly equal to those for TFPI-2, and staining levels lower than those for TFPI-2 were obtained for MMP-2 in all specimens, whereas for MMP-3 and MMP-9 they were lower in 85.7% and 57.1% of tissues, respectively.

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