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SULF2 overexpression positively regulates tumorigenicity of human prostate cancer cells.

Vicente CM, Lima MA, Nader HB, Toma L - J. Exp. Clin. Cancer Res. (2015)

Bottom Line: Transfection of DU-145 and PC3 prostate cancer cells with SULF2 resulted in increased viability, which did not occur with normal prostate cells.The effect was reverted by the knockdown of SULF2 using specific siRNAs.Detailed structural analysis of HS from cells overexpressing SULF2 showed a reduction of the trisulfated disaccharide UA(2S)-GlcNS(6S).

View Article: PubMed Central - PubMed

Affiliation: Departamento de Bioquímica, Disciplina de Biologia Molecular, Universidade Federal de São Paulo, UNIFESP, Rua Três de Maio, 100 - 4° andar, Vila Clementino, CEP 04044-020, São Paulo, SP, Brazil. carolmv@yahoo.com.

ABSTRACT

Background: SULF2 is a 6-O-endosulfatase which removes 6-O sulfate residues from N-glucosamine present on heparan sulfate (HS). The sulfation pattern of HS influences signaling events mediated by heparan sulfate proteoglycans (HSPGs) located on cell surface, which are critical for the interactions with growth factors and their receptors. Alterations in SULF2 expression have been identified in the context of several cancer types but its function in cancer is still unclear where the precise molecular mechanism involved has not been fully deciphered. To further investigate SULF2 role in tumorigenesis, we overexpressed such gene in prostate cancer cell lines.

Methods: The normal prostate epithelial cell line RWPE-1 and the prostate cancer cells DU-145, and PC3 were transfected with SULF2-expressing plasmid pcDNA3.1/Myc-His(-)-Hsulf-2. Transfected cells were then submitted to viability, migration and colony formation assays.

Results: Transfection of DU-145 and PC3 prostate cancer cells with SULF2 resulted in increased viability, which did not occur with normal prostate cells. The effect was reverted by the knockdown of SULF2 using specific siRNAs. Furthermore, forced expression of SULF2 augmented cell migration and colony formation in both prostate cell lines. Detailed structural analysis of HS from cells overexpressing SULF2 showed a reduction of the trisulfated disaccharide UA(2S)-GlcNS(6S). There was an increase in epithelial-mesenchymal transition markers and an increase in WNT signaling pathway.

Conclusions: These results indicate that SULF2 have a pro-tumorigenic effect in DU-145 and PC3 cancer cells, suggesting an important role of this enzyme in prostatic cancer metastasis.

No MeSH data available.


Related in: MedlinePlus

Forced expression of SULF2 on prostate cancer cells increased EMT markers. DU-145 (A) and PC3 (B) prostate cancer cell lines overexpressing SULF2 were stained with anti-CD44, anti-vimentin and anti-N-caderin antibodies and analyzed by flow cytometry, as described in methods. The graphics represent relative quantity of positively stained cells (C, D). *P ≤ 0.05.
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Fig6: Forced expression of SULF2 on prostate cancer cells increased EMT markers. DU-145 (A) and PC3 (B) prostate cancer cell lines overexpressing SULF2 were stained with anti-CD44, anti-vimentin and anti-N-caderin antibodies and analyzed by flow cytometry, as described in methods. The graphics represent relative quantity of positively stained cells (C, D). *P ≤ 0.05.

Mentions: The epithelial-mesenchymal transition (EMT) is a key developmental program that is often activated during cancer invasion and metastasis [39]. Recently, EMT markers have been found to confer malignant traits, such as motility, invasiveness and resistance to apoptosis [39]. Since we have observed an increase of these characteristics on prostate cancer cells with forced expression of SULF2, we decided to analyze some EMT markers in these cells. Thus, prostate cancer cells were immunostained for CD44, vimentin, and N-cadherin and the presence as well as their quantity analyzed by flow cytometry. Indeed, PC3 and DU-145 prostate cancer cells overexpressing SULF2 exhibited increased levels of CD44, vimentin, and N-cadherin (Figure 6). Hence, our results indicate that prostate cancer cells overexpressing SULF2 become more undifferentiated, which is in agreement with the increased cell growth and migration presented by them.Figure 6


SULF2 overexpression positively regulates tumorigenicity of human prostate cancer cells.

Vicente CM, Lima MA, Nader HB, Toma L - J. Exp. Clin. Cancer Res. (2015)

Forced expression of SULF2 on prostate cancer cells increased EMT markers. DU-145 (A) and PC3 (B) prostate cancer cell lines overexpressing SULF2 were stained with anti-CD44, anti-vimentin and anti-N-caderin antibodies and analyzed by flow cytometry, as described in methods. The graphics represent relative quantity of positively stained cells (C, D). *P ≤ 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4374423&req=5

Fig6: Forced expression of SULF2 on prostate cancer cells increased EMT markers. DU-145 (A) and PC3 (B) prostate cancer cell lines overexpressing SULF2 were stained with anti-CD44, anti-vimentin and anti-N-caderin antibodies and analyzed by flow cytometry, as described in methods. The graphics represent relative quantity of positively stained cells (C, D). *P ≤ 0.05.
Mentions: The epithelial-mesenchymal transition (EMT) is a key developmental program that is often activated during cancer invasion and metastasis [39]. Recently, EMT markers have been found to confer malignant traits, such as motility, invasiveness and resistance to apoptosis [39]. Since we have observed an increase of these characteristics on prostate cancer cells with forced expression of SULF2, we decided to analyze some EMT markers in these cells. Thus, prostate cancer cells were immunostained for CD44, vimentin, and N-cadherin and the presence as well as their quantity analyzed by flow cytometry. Indeed, PC3 and DU-145 prostate cancer cells overexpressing SULF2 exhibited increased levels of CD44, vimentin, and N-cadherin (Figure 6). Hence, our results indicate that prostate cancer cells overexpressing SULF2 become more undifferentiated, which is in agreement with the increased cell growth and migration presented by them.Figure 6

Bottom Line: Transfection of DU-145 and PC3 prostate cancer cells with SULF2 resulted in increased viability, which did not occur with normal prostate cells.The effect was reverted by the knockdown of SULF2 using specific siRNAs.Detailed structural analysis of HS from cells overexpressing SULF2 showed a reduction of the trisulfated disaccharide UA(2S)-GlcNS(6S).

View Article: PubMed Central - PubMed

Affiliation: Departamento de Bioquímica, Disciplina de Biologia Molecular, Universidade Federal de São Paulo, UNIFESP, Rua Três de Maio, 100 - 4° andar, Vila Clementino, CEP 04044-020, São Paulo, SP, Brazil. carolmv@yahoo.com.

ABSTRACT

Background: SULF2 is a 6-O-endosulfatase which removes 6-O sulfate residues from N-glucosamine present on heparan sulfate (HS). The sulfation pattern of HS influences signaling events mediated by heparan sulfate proteoglycans (HSPGs) located on cell surface, which are critical for the interactions with growth factors and their receptors. Alterations in SULF2 expression have been identified in the context of several cancer types but its function in cancer is still unclear where the precise molecular mechanism involved has not been fully deciphered. To further investigate SULF2 role in tumorigenesis, we overexpressed such gene in prostate cancer cell lines.

Methods: The normal prostate epithelial cell line RWPE-1 and the prostate cancer cells DU-145, and PC3 were transfected with SULF2-expressing plasmid pcDNA3.1/Myc-His(-)-Hsulf-2. Transfected cells were then submitted to viability, migration and colony formation assays.

Results: Transfection of DU-145 and PC3 prostate cancer cells with SULF2 resulted in increased viability, which did not occur with normal prostate cells. The effect was reverted by the knockdown of SULF2 using specific siRNAs. Furthermore, forced expression of SULF2 augmented cell migration and colony formation in both prostate cell lines. Detailed structural analysis of HS from cells overexpressing SULF2 showed a reduction of the trisulfated disaccharide UA(2S)-GlcNS(6S). There was an increase in epithelial-mesenchymal transition markers and an increase in WNT signaling pathway.

Conclusions: These results indicate that SULF2 have a pro-tumorigenic effect in DU-145 and PC3 cancer cells, suggesting an important role of this enzyme in prostatic cancer metastasis.

No MeSH data available.


Related in: MedlinePlus