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Discoidin domain receptor 1 contributes to tumorigenesis through modulation of TGFBI expression.

Rudra-Ganguly N, Lowe C, Mattie M, Chang MS, Satpayev D, Verlinsky A, An Z, Hu L, Yang P, Challita-Eid P, Stover DR, Pereira DS - PLoS ONE (2014)

Bottom Line: Although DDR1 is expressed in many normal tissues, upregulated expression of DDR1 in a variety of human cancers such as lung, colon and brain cancers is known to be associated with poor prognosis.Together, these data suggest that DDR1 expression level influences tumor growth in part via modulation of TGFBI expression.The reciprocal expression of DDR1 and TGFBI may help to elucidate the contribution of DDR1 in tumorigenesis and TGFBI may also be used as a biomarker for the therapeutic development of DDR1 specific inhibitors.

View Article: PubMed Central - PubMed

Affiliation: Agensys Inc., an affiliate of Astellas Pharma Inc, Santa Monica, CA, United States of America.

ABSTRACT
Discoidin domain receptor 1 (DDR1) is a member of the receptor tyrosine kinase family. The receptor is activated upon binding to its ligand, collagen, and plays a crucial role in many fundamental processes such as cell differentiation, adhesion, migration and invasion. Although DDR1 is expressed in many normal tissues, upregulated expression of DDR1 in a variety of human cancers such as lung, colon and brain cancers is known to be associated with poor prognosis. Using shRNA silencing, we assessed the oncogenic potential of DDR1. DDR1 knockdown impaired tumor cell proliferation and migration in vitro and tumor growth in vivo. Microarray analysis of tumor cells demonstrated upregulation of TGFBI expression upon DDR1 knockdown, which was subsequently confirmed at the protein level. TGFBI is a TGFβ-induced extracellular matrix protein secreted by the tumor cells and is known to act either as a tumor promoter or tumor suppressor, depending on the tumor environment. Here, we show that exogenous addition of recombinant TGFBI to BXPC3 tumor cells inhibited clonogenic growth and migration, thus recapitulating the phenotypic effect observed from DDR1 silencing. BXPC3 tumor xenografts demonstrated reduced growth with DDR1 knockdown, and the same xenograft tumors exhibited an increase in TGFBI expression level. Together, these data suggest that DDR1 expression level influences tumor growth in part via modulation of TGFBI expression. The reciprocal expression of DDR1 and TGFBI may help to elucidate the contribution of DDR1 in tumorigenesis and TGFBI may also be used as a biomarker for the therapeutic development of DDR1 specific inhibitors.

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Efficacy of shRNA targeting DDR1 on BXPC3 tumor growth.(a) BXPC3 cells untreated (•), NT shRNA (▪) and DDR1 shRNA5 (Δ) were implanted in ICR-SCID mice and tumor growth was monitored up to 28 days. (b) On day 28, when compared to untreated and NT shRNA groups, the tumors in DDR1 shRNA5 group was the smallest as measured by the tumor volume (N = 10). The calculated percent inhibition in tumor growth in DDR1 shRNA group was 50.7% (***, p<0.0001) and 43.2% (**, p<0.0052) when compared to parental and NT shRNA groups, respectively. Tumors were harvested and DDR1 expression was confirmed by (c) immunohistochemistry and (d) western blot.
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pone-0111515-g003: Efficacy of shRNA targeting DDR1 on BXPC3 tumor growth.(a) BXPC3 cells untreated (•), NT shRNA (▪) and DDR1 shRNA5 (Δ) were implanted in ICR-SCID mice and tumor growth was monitored up to 28 days. (b) On day 28, when compared to untreated and NT shRNA groups, the tumors in DDR1 shRNA5 group was the smallest as measured by the tumor volume (N = 10). The calculated percent inhibition in tumor growth in DDR1 shRNA group was 50.7% (***, p<0.0001) and 43.2% (**, p<0.0052) when compared to parental and NT shRNA groups, respectively. Tumors were harvested and DDR1 expression was confirmed by (c) immunohistochemistry and (d) western blot.

Mentions: To evaluate whether DDR1 expression is required for tumor cell growth in vivo, we examined the effect of DDR1 knockdown on the growth of BXPC3 tumor xenografts in mice. Tumor growth monitored for 25 days showed that DDR1 knockdown resulted in approximate 50% tumor growth inhibition compared to the control groups (Fig. 3a). Statistical analysis of tumor size data on day 25 (Fig. 3b) indicated smaller tumors in the DDR1 shRNA5 group than those in the parental (p<0.0001) or NT shRNA group (p<0.01). To confirm DDR1 protein expression level in vivo, we evaluated DDR1 expression by immunohistochemistry and western blot in tumors at the end of the study. (Fig. 3c and 3d) Both analyses confirmed reduced DDR1 expression in the tumors from the shRNA5 group compared to the tumors from either the parental or the NT shRNA group. The loss of DDR1 expression in tumors might contribute towards attenuated tumor growth.


Discoidin domain receptor 1 contributes to tumorigenesis through modulation of TGFBI expression.

Rudra-Ganguly N, Lowe C, Mattie M, Chang MS, Satpayev D, Verlinsky A, An Z, Hu L, Yang P, Challita-Eid P, Stover DR, Pereira DS - PLoS ONE (2014)

Efficacy of shRNA targeting DDR1 on BXPC3 tumor growth.(a) BXPC3 cells untreated (•), NT shRNA (▪) and DDR1 shRNA5 (Δ) were implanted in ICR-SCID mice and tumor growth was monitored up to 28 days. (b) On day 28, when compared to untreated and NT shRNA groups, the tumors in DDR1 shRNA5 group was the smallest as measured by the tumor volume (N = 10). The calculated percent inhibition in tumor growth in DDR1 shRNA group was 50.7% (***, p<0.0001) and 43.2% (**, p<0.0052) when compared to parental and NT shRNA groups, respectively. Tumors were harvested and DDR1 expression was confirmed by (c) immunohistochemistry and (d) western blot.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4219757&req=5

pone-0111515-g003: Efficacy of shRNA targeting DDR1 on BXPC3 tumor growth.(a) BXPC3 cells untreated (•), NT shRNA (▪) and DDR1 shRNA5 (Δ) were implanted in ICR-SCID mice and tumor growth was monitored up to 28 days. (b) On day 28, when compared to untreated and NT shRNA groups, the tumors in DDR1 shRNA5 group was the smallest as measured by the tumor volume (N = 10). The calculated percent inhibition in tumor growth in DDR1 shRNA group was 50.7% (***, p<0.0001) and 43.2% (**, p<0.0052) when compared to parental and NT shRNA groups, respectively. Tumors were harvested and DDR1 expression was confirmed by (c) immunohistochemistry and (d) western blot.
Mentions: To evaluate whether DDR1 expression is required for tumor cell growth in vivo, we examined the effect of DDR1 knockdown on the growth of BXPC3 tumor xenografts in mice. Tumor growth monitored for 25 days showed that DDR1 knockdown resulted in approximate 50% tumor growth inhibition compared to the control groups (Fig. 3a). Statistical analysis of tumor size data on day 25 (Fig. 3b) indicated smaller tumors in the DDR1 shRNA5 group than those in the parental (p<0.0001) or NT shRNA group (p<0.01). To confirm DDR1 protein expression level in vivo, we evaluated DDR1 expression by immunohistochemistry and western blot in tumors at the end of the study. (Fig. 3c and 3d) Both analyses confirmed reduced DDR1 expression in the tumors from the shRNA5 group compared to the tumors from either the parental or the NT shRNA group. The loss of DDR1 expression in tumors might contribute towards attenuated tumor growth.

Bottom Line: Although DDR1 is expressed in many normal tissues, upregulated expression of DDR1 in a variety of human cancers such as lung, colon and brain cancers is known to be associated with poor prognosis.Together, these data suggest that DDR1 expression level influences tumor growth in part via modulation of TGFBI expression.The reciprocal expression of DDR1 and TGFBI may help to elucidate the contribution of DDR1 in tumorigenesis and TGFBI may also be used as a biomarker for the therapeutic development of DDR1 specific inhibitors.

View Article: PubMed Central - PubMed

Affiliation: Agensys Inc., an affiliate of Astellas Pharma Inc, Santa Monica, CA, United States of America.

ABSTRACT
Discoidin domain receptor 1 (DDR1) is a member of the receptor tyrosine kinase family. The receptor is activated upon binding to its ligand, collagen, and plays a crucial role in many fundamental processes such as cell differentiation, adhesion, migration and invasion. Although DDR1 is expressed in many normal tissues, upregulated expression of DDR1 in a variety of human cancers such as lung, colon and brain cancers is known to be associated with poor prognosis. Using shRNA silencing, we assessed the oncogenic potential of DDR1. DDR1 knockdown impaired tumor cell proliferation and migration in vitro and tumor growth in vivo. Microarray analysis of tumor cells demonstrated upregulation of TGFBI expression upon DDR1 knockdown, which was subsequently confirmed at the protein level. TGFBI is a TGFβ-induced extracellular matrix protein secreted by the tumor cells and is known to act either as a tumor promoter or tumor suppressor, depending on the tumor environment. Here, we show that exogenous addition of recombinant TGFBI to BXPC3 tumor cells inhibited clonogenic growth and migration, thus recapitulating the phenotypic effect observed from DDR1 silencing. BXPC3 tumor xenografts demonstrated reduced growth with DDR1 knockdown, and the same xenograft tumors exhibited an increase in TGFBI expression level. Together, these data suggest that DDR1 expression level influences tumor growth in part via modulation of TGFBI expression. The reciprocal expression of DDR1 and TGFBI may help to elucidate the contribution of DDR1 in tumorigenesis and TGFBI may also be used as a biomarker for the therapeutic development of DDR1 specific inhibitors.

Show MeSH
Related in: MedlinePlus