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Polypeptide N-acetylgalactosaminyltransferase 2 regulates cellular metastasis-associated behavior in gastric cancer.

Hua D, Shen L, Xu L, Jiang Z, Zhou Y, Yue A, Zou S, Cheng Z, Wu S - Int. J. Mol. Med. (2012)

Bottom Line: We found that cell proliferation, adhesion and invasion were decreased in ppGalNAc-T2 overexpressed cells but increased in ppGalNAc-T2 downregulated cells.However, it did not exhibit any apparent correlation with MMP-14 expression levels.Our data show the effect of ppGalNAc-T2 on proliferation, adhesion or invasion of SGC7901 gastric cancer cells, suggesting that ppGalNAc-T2 may exert anti-proliferative and anti-metastatic activity through the decrease of MMP-2 and TGF-β1.

View Article: PubMed Central - PubMed

Affiliation: Fourth Affiliated Hospital of Soochow University, Wuxi, Jiangsu 214062, P.R. China.

ABSTRACT
Aberrant glycosylation of cell surface glycoprotein due to specific alterations of glycosyltransferase activity is usually associated with invasion and metastasis of cancer, particularly of gastric carcinomas. Polypeptide N-acetylgalactosaminyltransferase 2 (ppGalNAc-T2), which catalyzes initiation of mucin-type O-glycosylation, is also involved in tumor migration and invasion. However, a comprehensive understanding of how ppGalNAc-T2 correlates with the metastasic potential of human gastric cancer is not currently available. In the present study, ppGalNAc-T2 was detected in a variety of human poorly differentiated tumor cells, and expression appeared to be higher in SGC7901 gastric cancer cells. In addition, we investigated the potential effects of ppGalNAc-T2 on growth and metastasis-associated behavior in SGC7901 cells after stable transfection with ppGalNAc-T2 sense and antisense vectors. We found that cell proliferation, adhesion and invasion were decreased in ppGalNAc-T2 overexpressed cells but increased in ppGalNAc-T2 downregulated cells. Therefore, we attempted to clarify the mechanisms underlying the anti-metastatic activities of ppGalNAc-T2. Further investigation indicated that overexpression of ppGalNAc-T2 is involved in the inhibition of matrix metalloproteinase (MMP)-2 expression at both the protein and mRNA levels, which may be associated with ppGalNAc-T2 suppressing the expression of transforming growth factor (TGF)-β1. However, it did not exhibit any apparent correlation with MMP-14 expression levels. Our data show the effect of ppGalNAc-T2 on proliferation, adhesion or invasion of SGC7901 gastric cancer cells, suggesting that ppGalNAc-T2 may exert anti-proliferative and anti-metastatic activity through the decrease of MMP-2 and TGF-β1. These results indicate that ppGalNAc‑T2 may be used as a novel therapeutic target for human gastric cancer treatment.

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Expression levels of MMP-2, MMP-14 and TGF-β1 in different SGC7901 clones ofstably transfected cells, including untransfected SGC7901 cells, SGC7901 cells stablyoverexpressing ppGalNAc-T2 (SGC7901-T2s), SGC7901 cells with downregulated expression ofppGalNAc-T2 (SGC7901-T2as) or with empty vector as control. β-actin was used asan internal control for loading. ppGalNAc-T2 expression in different SGC7901 cells wasanalyzed by (A) RT-PCR and (C) western blot analysis. (B and D) Band intensity wasquantified using densitometry and normalized to β-actin band intensity. 1,untreated SGC7901 cells; 2, control group; 3, SGC7901-T2s group; and 4, SGC7901-T2asgroup. (*P<0.05, **P>0.05compared to the untreated group).
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f7-ijmm-30-06-1267: Expression levels of MMP-2, MMP-14 and TGF-β1 in different SGC7901 clones ofstably transfected cells, including untransfected SGC7901 cells, SGC7901 cells stablyoverexpressing ppGalNAc-T2 (SGC7901-T2s), SGC7901 cells with downregulated expression ofppGalNAc-T2 (SGC7901-T2as) or with empty vector as control. β-actin was used asan internal control for loading. ppGalNAc-T2 expression in different SGC7901 cells wasanalyzed by (A) RT-PCR and (C) western blot analysis. (B and D) Band intensity wasquantified using densitometry and normalized to β-actin band intensity. 1,untreated SGC7901 cells; 2, control group; 3, SGC7901-T2s group; and 4, SGC7901-T2asgroup. (*P<0.05, **P>0.05compared to the untreated group).

Mentions: Among the MMP family that has been identified, MMP-2 is considered a key enzyme since itis responsible for degradation of the ECM. Meanwhile, MMP-2 activity can be activated byMMP-14, and this activity may be involved in tumor invasion and metastasis. Therefore, toinvestigate whether the metastasic inhibitory effect of ppGalNAc-T2 resulted from thesuppression of MMP-2 and MMP-14 expression, MMP-2 and MMP-14 mRNA and protein levels weremeasured. Using RT-PCR, we found that the expression of MMP-2 at the mRNA level was lowerin the SGC7901-T2s group than in the SGC7901-T2as group (P<0.05), and there was nodifference between untreated and control group cells (P>0.05) (Fig. 7A and B). However, there was noevident change on the mRNA transcriptional expression of MMP-14.


Polypeptide N-acetylgalactosaminyltransferase 2 regulates cellular metastasis-associated behavior in gastric cancer.

Hua D, Shen L, Xu L, Jiang Z, Zhou Y, Yue A, Zou S, Cheng Z, Wu S - Int. J. Mol. Med. (2012)

Expression levels of MMP-2, MMP-14 and TGF-β1 in different SGC7901 clones ofstably transfected cells, including untransfected SGC7901 cells, SGC7901 cells stablyoverexpressing ppGalNAc-T2 (SGC7901-T2s), SGC7901 cells with downregulated expression ofppGalNAc-T2 (SGC7901-T2as) or with empty vector as control. β-actin was used asan internal control for loading. ppGalNAc-T2 expression in different SGC7901 cells wasanalyzed by (A) RT-PCR and (C) western blot analysis. (B and D) Band intensity wasquantified using densitometry and normalized to β-actin band intensity. 1,untreated SGC7901 cells; 2, control group; 3, SGC7901-T2s group; and 4, SGC7901-T2asgroup. (*P<0.05, **P>0.05compared to the untreated group).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4042861&req=5

f7-ijmm-30-06-1267: Expression levels of MMP-2, MMP-14 and TGF-β1 in different SGC7901 clones ofstably transfected cells, including untransfected SGC7901 cells, SGC7901 cells stablyoverexpressing ppGalNAc-T2 (SGC7901-T2s), SGC7901 cells with downregulated expression ofppGalNAc-T2 (SGC7901-T2as) or with empty vector as control. β-actin was used asan internal control for loading. ppGalNAc-T2 expression in different SGC7901 cells wasanalyzed by (A) RT-PCR and (C) western blot analysis. (B and D) Band intensity wasquantified using densitometry and normalized to β-actin band intensity. 1,untreated SGC7901 cells; 2, control group; 3, SGC7901-T2s group; and 4, SGC7901-T2asgroup. (*P<0.05, **P>0.05compared to the untreated group).
Mentions: Among the MMP family that has been identified, MMP-2 is considered a key enzyme since itis responsible for degradation of the ECM. Meanwhile, MMP-2 activity can be activated byMMP-14, and this activity may be involved in tumor invasion and metastasis. Therefore, toinvestigate whether the metastasic inhibitory effect of ppGalNAc-T2 resulted from thesuppression of MMP-2 and MMP-14 expression, MMP-2 and MMP-14 mRNA and protein levels weremeasured. Using RT-PCR, we found that the expression of MMP-2 at the mRNA level was lowerin the SGC7901-T2s group than in the SGC7901-T2as group (P<0.05), and there was nodifference between untreated and control group cells (P>0.05) (Fig. 7A and B). However, there was noevident change on the mRNA transcriptional expression of MMP-14.

Bottom Line: We found that cell proliferation, adhesion and invasion were decreased in ppGalNAc-T2 overexpressed cells but increased in ppGalNAc-T2 downregulated cells.However, it did not exhibit any apparent correlation with MMP-14 expression levels.Our data show the effect of ppGalNAc-T2 on proliferation, adhesion or invasion of SGC7901 gastric cancer cells, suggesting that ppGalNAc-T2 may exert anti-proliferative and anti-metastatic activity through the decrease of MMP-2 and TGF-β1.

View Article: PubMed Central - PubMed

Affiliation: Fourth Affiliated Hospital of Soochow University, Wuxi, Jiangsu 214062, P.R. China.

ABSTRACT
Aberrant glycosylation of cell surface glycoprotein due to specific alterations of glycosyltransferase activity is usually associated with invasion and metastasis of cancer, particularly of gastric carcinomas. Polypeptide N-acetylgalactosaminyltransferase 2 (ppGalNAc-T2), which catalyzes initiation of mucin-type O-glycosylation, is also involved in tumor migration and invasion. However, a comprehensive understanding of how ppGalNAc-T2 correlates with the metastasic potential of human gastric cancer is not currently available. In the present study, ppGalNAc-T2 was detected in a variety of human poorly differentiated tumor cells, and expression appeared to be higher in SGC7901 gastric cancer cells. In addition, we investigated the potential effects of ppGalNAc-T2 on growth and metastasis-associated behavior in SGC7901 cells after stable transfection with ppGalNAc-T2 sense and antisense vectors. We found that cell proliferation, adhesion and invasion were decreased in ppGalNAc-T2 overexpressed cells but increased in ppGalNAc-T2 downregulated cells. Therefore, we attempted to clarify the mechanisms underlying the anti-metastatic activities of ppGalNAc-T2. Further investigation indicated that overexpression of ppGalNAc-T2 is involved in the inhibition of matrix metalloproteinase (MMP)-2 expression at both the protein and mRNA levels, which may be associated with ppGalNAc-T2 suppressing the expression of transforming growth factor (TGF)-β1. However, it did not exhibit any apparent correlation with MMP-14 expression levels. Our data show the effect of ppGalNAc-T2 on proliferation, adhesion or invasion of SGC7901 gastric cancer cells, suggesting that ppGalNAc-T2 may exert anti-proliferative and anti-metastatic activity through the decrease of MMP-2 and TGF-β1. These results indicate that ppGalNAc‑T2 may be used as a novel therapeutic target for human gastric cancer treatment.

Show MeSH
Related in: MedlinePlus