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Sulf-2, a heparan sulfate endosulfatase, promotes human lung carcinogenesis.

Lemjabbar-Alaoui H, van Zante A, Singer MS, Xue Q, Wang YQ, Tsay D, He B, Jablons DM, Rosen SD - Oncogene (2009)

Bottom Line: We found induction of SULF2 transcripts and Sulf-2 protein in human lung adenocarcinoma and squamous cell carcinoma, the two major classes of non-small-cell lung carcinomas (NSCLCs).We confirmed widespread Sulf-2 protein expression in tumor cells of 10/10 surgical specimens of human lung squamous carcinomas.Our findings support an essential role for Sulf-2 in lung cancer, the leading cancer killer.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, University of California, San Francisco, CA 94143, USA.

ABSTRACT
Heparan sulfate (HS) proteoglycans (HSPGs) bind to multiple growth factors/morphogens and regulate their signaling. 6-O-sulfation (6S) of glucosamine within HS chains is critical for many of these ligand interactions. Sulf-1 and Sulf-2, which are extracellular neutral-pH sulfatases, provide a novel post-synthetic mechanism for regulation of HSPG function by removing 6S from intact HS chains. The Sulfs can thereby modulate several signaling pathways, including the promotion of Wnt signaling. We found induction of SULF2 transcripts and Sulf-2 protein in human lung adenocarcinoma and squamous cell carcinoma, the two major classes of non-small-cell lung carcinomas (NSCLCs). We confirmed widespread Sulf-2 protein expression in tumor cells of 10/10 surgical specimens of human lung squamous carcinomas. We studied five Sulf-2(+) NSCLC cell lines, including two, which were derived by cigarette-smoke transformation of bronchial epithelial cells. shRNA-mediated Sulf-2 knockdown in these lines caused an increase in 6S on their cell surface and in parallel reversed their transformed phenotype in vitro, eliminated autocrine Wnt signaling and strongly blunted xenograft tumor formation in nude mice. Conversely, forced Sulf-2 expression in non-malignant bronchial epithelial cells produced a partially transformed phenotype. Our findings support an essential role for Sulf-2 in lung cancer, the leading cancer killer.

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Effects of Sulf-2 overexpression in non-malignant bronchial epithelial cells: (a) BEAS2B and H140 were transduced with either Sulf2 or inactive Sulf-2 (S2ΔCC). Immunoblotting of Sulf-2 protein in CM (Upper) and cell lysates (Middle) of transduced cells as compared to Sulf-2+ cells (H292, Calu-6 and P-ST) with b-actin as loading control (Lower). (b) Staining of parental and transduced cells with RB4CD12 or control antibody, BEAS2B (left), H140 (right). (c) Healing of scratch wounds by parental and transduced cells. Scale bar = 76μm. (d) Growth of parental and transduced cells by Cell Titer Blue assay. (e) Soft agar colony formation by parental and transduced cells after 21 days of culture. (f) 9 clones of Sulf-2 expressing BEAS2B cells were generated from colonies in soft agar. Apoptosis was determined for the clones, parentals, and transduced populations by measuring staining with APC conjugated-Annexin V. The values shown in graphs are means ± SDs for 3 independent determinations (* indicates p< 0.05).
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Figure 4: Effects of Sulf-2 overexpression in non-malignant bronchial epithelial cells: (a) BEAS2B and H140 were transduced with either Sulf2 or inactive Sulf-2 (S2ΔCC). Immunoblotting of Sulf-2 protein in CM (Upper) and cell lysates (Middle) of transduced cells as compared to Sulf-2+ cells (H292, Calu-6 and P-ST) with b-actin as loading control (Lower). (b) Staining of parental and transduced cells with RB4CD12 or control antibody, BEAS2B (left), H140 (right). (c) Healing of scratch wounds by parental and transduced cells. Scale bar = 76μm. (d) Growth of parental and transduced cells by Cell Titer Blue assay. (e) Soft agar colony formation by parental and transduced cells after 21 days of culture. (f) 9 clones of Sulf-2 expressing BEAS2B cells were generated from colonies in soft agar. Apoptosis was determined for the clones, parentals, and transduced populations by measuring staining with APC conjugated-Annexin V. The values shown in graphs are means ± SDs for 3 independent determinations (* indicates p< 0.05).

Mentions: These loss-of-function findings suggested that Sulf-2 may have a transforming capability in lung epithelial cells. To examine the functional consequences of Sulf-2 expression in lung cells, we overexpressed either active or inactive Sulf-2 (S2ΔCC) in two non-malignant human bronchial epithelial cell lines (BEAS2B and 16HBE14o-), both of which are negative for the Sulfs. Transduction resulted in Sulf-2 expression levels comparable to those found in high-expressing lung cancer cell lines (Fig. 4a).


Sulf-2, a heparan sulfate endosulfatase, promotes human lung carcinogenesis.

Lemjabbar-Alaoui H, van Zante A, Singer MS, Xue Q, Wang YQ, Tsay D, He B, Jablons DM, Rosen SD - Oncogene (2009)

Effects of Sulf-2 overexpression in non-malignant bronchial epithelial cells: (a) BEAS2B and H140 were transduced with either Sulf2 or inactive Sulf-2 (S2ΔCC). Immunoblotting of Sulf-2 protein in CM (Upper) and cell lysates (Middle) of transduced cells as compared to Sulf-2+ cells (H292, Calu-6 and P-ST) with b-actin as loading control (Lower). (b) Staining of parental and transduced cells with RB4CD12 or control antibody, BEAS2B (left), H140 (right). (c) Healing of scratch wounds by parental and transduced cells. Scale bar = 76μm. (d) Growth of parental and transduced cells by Cell Titer Blue assay. (e) Soft agar colony formation by parental and transduced cells after 21 days of culture. (f) 9 clones of Sulf-2 expressing BEAS2B cells were generated from colonies in soft agar. Apoptosis was determined for the clones, parentals, and transduced populations by measuring staining with APC conjugated-Annexin V. The values shown in graphs are means ± SDs for 3 independent determinations (* indicates p< 0.05).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2818095&req=5

Figure 4: Effects of Sulf-2 overexpression in non-malignant bronchial epithelial cells: (a) BEAS2B and H140 were transduced with either Sulf2 or inactive Sulf-2 (S2ΔCC). Immunoblotting of Sulf-2 protein in CM (Upper) and cell lysates (Middle) of transduced cells as compared to Sulf-2+ cells (H292, Calu-6 and P-ST) with b-actin as loading control (Lower). (b) Staining of parental and transduced cells with RB4CD12 or control antibody, BEAS2B (left), H140 (right). (c) Healing of scratch wounds by parental and transduced cells. Scale bar = 76μm. (d) Growth of parental and transduced cells by Cell Titer Blue assay. (e) Soft agar colony formation by parental and transduced cells after 21 days of culture. (f) 9 clones of Sulf-2 expressing BEAS2B cells were generated from colonies in soft agar. Apoptosis was determined for the clones, parentals, and transduced populations by measuring staining with APC conjugated-Annexin V. The values shown in graphs are means ± SDs for 3 independent determinations (* indicates p< 0.05).
Mentions: These loss-of-function findings suggested that Sulf-2 may have a transforming capability in lung epithelial cells. To examine the functional consequences of Sulf-2 expression in lung cells, we overexpressed either active or inactive Sulf-2 (S2ΔCC) in two non-malignant human bronchial epithelial cell lines (BEAS2B and 16HBE14o-), both of which are negative for the Sulfs. Transduction resulted in Sulf-2 expression levels comparable to those found in high-expressing lung cancer cell lines (Fig. 4a).

Bottom Line: We found induction of SULF2 transcripts and Sulf-2 protein in human lung adenocarcinoma and squamous cell carcinoma, the two major classes of non-small-cell lung carcinomas (NSCLCs).We confirmed widespread Sulf-2 protein expression in tumor cells of 10/10 surgical specimens of human lung squamous carcinomas.Our findings support an essential role for Sulf-2 in lung cancer, the leading cancer killer.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, University of California, San Francisco, CA 94143, USA.

ABSTRACT
Heparan sulfate (HS) proteoglycans (HSPGs) bind to multiple growth factors/morphogens and regulate their signaling. 6-O-sulfation (6S) of glucosamine within HS chains is critical for many of these ligand interactions. Sulf-1 and Sulf-2, which are extracellular neutral-pH sulfatases, provide a novel post-synthetic mechanism for regulation of HSPG function by removing 6S from intact HS chains. The Sulfs can thereby modulate several signaling pathways, including the promotion of Wnt signaling. We found induction of SULF2 transcripts and Sulf-2 protein in human lung adenocarcinoma and squamous cell carcinoma, the two major classes of non-small-cell lung carcinomas (NSCLCs). We confirmed widespread Sulf-2 protein expression in tumor cells of 10/10 surgical specimens of human lung squamous carcinomas. We studied five Sulf-2(+) NSCLC cell lines, including two, which were derived by cigarette-smoke transformation of bronchial epithelial cells. shRNA-mediated Sulf-2 knockdown in these lines caused an increase in 6S on their cell surface and in parallel reversed their transformed phenotype in vitro, eliminated autocrine Wnt signaling and strongly blunted xenograft tumor formation in nude mice. Conversely, forced Sulf-2 expression in non-malignant bronchial epithelial cells produced a partially transformed phenotype. Our findings support an essential role for Sulf-2 in lung cancer, the leading cancer killer.

Show MeSH
Related in: MedlinePlus