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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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Sulf-2 protein expression in NSCLC tumors: representative sections of benign lung and squamous cell carcinoma were stained with hematoxylin and eosin (H&E) and adjacent serial sections were stained with anti-Sulf-2 antibody (2B4). (a) Normal lung, H&E. (b) Normal lung stained with 2B4. (c) Squamous cell carcinoma, H&E. (d) Squamous cell carcinoma stained with 2B4 antibody demonstrates islands of tumor cells strongly positive for Sulf-2 surrounded by weakly staining desmoplastic stroma. Panels a, b, c and d are low-power micrographs (100X, scale bar = 500μm). (e and f) High-power micrographs of squamous cell carcinoma stained with 2B4 antibody. Panel f shows staining of tumor-associated stromal cells with 2B4 antibody (400X, scale bar = 100μm).
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Figure 2: Sulf-2 protein expression in NSCLC tumors: representative sections of benign lung and squamous cell carcinoma were stained with hematoxylin and eosin (H&E) and adjacent serial sections were stained with anti-Sulf-2 antibody (2B4). (a) Normal lung, H&E. (b) Normal lung stained with 2B4. (c) Squamous cell carcinoma, H&E. (d) Squamous cell carcinoma stained with 2B4 antibody demonstrates islands of tumor cells strongly positive for Sulf-2 surrounded by weakly staining desmoplastic stroma. Panels a, b, c and d are low-power micrographs (100X, scale bar = 500μm). (e and f) High-power micrographs of squamous cell carcinoma stained with 2B4 antibody. Panel f shows staining of tumor-associated stromal cells with 2B4 antibody (400X, scale bar = 100μm).

Mentions: We performed immunocytochemistry for Sulf-2 in NSCLC tumors (ten cases each of squamous cell carcinoma and adenocarcinoma) selected from archived surgical specimens (Supplementary Table 2). Normal lung tissue from the same specimens served as controls. Paraffin sections were stained with a newly developed Sulf-2 mAb (Supplementary Fig. 3). All ten cases of squamous cell carcinoma showed some degree of staining for Sulf-2 in the tumor cells. The staining was patchy and accentuated in the cells at the periphery of nests (Fig. 2). The percentage of positive cells was variable and the intensity of staining ranged from faint to intense (Supplementary Table 2). No correlation was found between tumor differentiation and the percentage of positive cells or intensity of staining. In striking contrast, there was no staining of adenocarcinoma cells in any of the cases. This was surprising since three of the NSCLC cell lines with SULF2 expression (Calu-3, Calu-6, and A549) could be classified as adenocarcinomas (Supplementary Table 1). However, both adenocarcinomas and squamous cell carcinomas showed conspicuous staining of tumor stroma, including spindle-shaped cells and endothelial cells of blood vessels. In control tissue distant from the tumors, endothelial cell staining was also seen (Fig. 2b), but normal airway epithelium, except for rare basal cells, was negative for Sulf-2.


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)

Sulf-2 protein expression in NSCLC tumors: representative sections of benign lung and squamous cell carcinoma were stained with hematoxylin and eosin (H&E) and adjacent serial sections were stained with anti-Sulf-2 antibody (2B4). (a) Normal lung, H&E. (b) Normal lung stained with 2B4. (c) Squamous cell carcinoma, H&E. (d) Squamous cell carcinoma stained with 2B4 antibody demonstrates islands of tumor cells strongly positive for Sulf-2 surrounded by weakly staining desmoplastic stroma. Panels a, b, c and d are low-power micrographs (100X, scale bar = 500μm). (e and f) High-power micrographs of squamous cell carcinoma stained with 2B4 antibody. Panel f shows staining of tumor-associated stromal cells with 2B4 antibody (400X, scale bar = 100μm).
© Copyright Policy
Related In: Results  -  Collection

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

Figure 2: Sulf-2 protein expression in NSCLC tumors: representative sections of benign lung and squamous cell carcinoma were stained with hematoxylin and eosin (H&E) and adjacent serial sections were stained with anti-Sulf-2 antibody (2B4). (a) Normal lung, H&E. (b) Normal lung stained with 2B4. (c) Squamous cell carcinoma, H&E. (d) Squamous cell carcinoma stained with 2B4 antibody demonstrates islands of tumor cells strongly positive for Sulf-2 surrounded by weakly staining desmoplastic stroma. Panels a, b, c and d are low-power micrographs (100X, scale bar = 500μm). (e and f) High-power micrographs of squamous cell carcinoma stained with 2B4 antibody. Panel f shows staining of tumor-associated stromal cells with 2B4 antibody (400X, scale bar = 100μm).
Mentions: We performed immunocytochemistry for Sulf-2 in NSCLC tumors (ten cases each of squamous cell carcinoma and adenocarcinoma) selected from archived surgical specimens (Supplementary Table 2). Normal lung tissue from the same specimens served as controls. Paraffin sections were stained with a newly developed Sulf-2 mAb (Supplementary Fig. 3). All ten cases of squamous cell carcinoma showed some degree of staining for Sulf-2 in the tumor cells. The staining was patchy and accentuated in the cells at the periphery of nests (Fig. 2). The percentage of positive cells was variable and the intensity of staining ranged from faint to intense (Supplementary Table 2). No correlation was found between tumor differentiation and the percentage of positive cells or intensity of staining. In striking contrast, there was no staining of adenocarcinoma cells in any of the cases. This was surprising since three of the NSCLC cell lines with SULF2 expression (Calu-3, Calu-6, and A549) could be classified as adenocarcinomas (Supplementary Table 1). However, both adenocarcinomas and squamous cell carcinomas showed conspicuous staining of tumor stroma, including spindle-shaped cells and endothelial cells of blood vessels. In control tissue distant from the tumors, endothelial cell staining was also seen (Fig. 2b), but normal airway epithelium, except for rare basal cells, was negative for Sulf-2.

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