Limits...
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.

Show MeSH

Related in: MedlinePlus

Expression levels of ppGalNAc-T2 in different SGC7901 clones of stably transfectedcells, including untransfected SGC7901 cells, SGC7901 cells stably overexpressingppGalNAc-T2 (SGC7901-T2s), SGC7901 cells with downregulated expression of ppGalNAc-T2(SGC7901-T2as) or with empty vector as control. β-actin was used as an internalcontrol for loading. ppGalNAc-T2 expression in different SGC7901 cells was analysed by(A) RT-PCR and (C) western blot analysis. (B and D) Band intensity was quantified usingdensitometry and normalized to β-actin band intensity. 1, Untreated SGC7901cells; 2, control group; 3, SGC7901-T2s group; and 4, SGC7901-T2as group.(*P<0.05, **P>0.05compared to the untreated group).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4042861&req=5

f3-ijmm-30-06-1267: Expression levels of ppGalNAc-T2 in different SGC7901 clones of stably transfectedcells, including untransfected SGC7901 cells, SGC7901 cells stably overexpressingppGalNAc-T2 (SGC7901-T2s), SGC7901 cells with downregulated expression of ppGalNAc-T2(SGC7901-T2as) or with empty vector as control. β-actin was used as an internalcontrol for loading. ppGalNAc-T2 expression in different SGC7901 cells was analysed by(A) RT-PCR and (C) western blot analysis. (B and D) Band intensity was quantified usingdensitometry and normalized to β-actin band intensity. 1, Untreated SGC7901cells; 2, control group; 3, SGC7901-T2s group; and 4, SGC7901-T2as group.(*P<0.05, **P>0.05compared to the untreated group).

Mentions: To further explore the role of ppGalNAc-T2 in gastric cancer, SGC7901 cells were used toreconstitute the expression of ppGalNAc-T2 by stable overexpression (SGC7901-T2s) ordownregulation (SGC7901-T2as) of ppGalNAc-T2. Transfection efficiency was measured usingfluorescence microscopy to detect expression of the plasmid-encoded eGFP gene (Fig. 2). Then, the ppGalNAc-T2 mRNA andprotein levels in the SGC7901 cells were measured by RT-PCR and western blot analysis,respectively. When compared with untreated cells, ppGalNAc-T2 transcripts were increasedin the pEGFP-C1-ppGalNAc-T2 sense vector transfected cells (P<0.05) (Fig. 3A and B). Consistent with theRT-PCR results, the expression of the ppGalNAc-T2 protein was clearly increased in thisgroup (P<0.05) (Fig. 3C andD). Furthermore, ppGalNAc-T2 mRNA and protein expression was suppressed in theSGC7901-T2as group when compared to the untreated group (P<0.05), while nodifference was found between the control group and the untreated cells (P>0.05).The above results indicate the successful construction of the ppGalNAc-T2 overexpressionor downregulation cell lines. These stable cell lines can be effectively used to furtherexamine the role of ppGalNAc-T2.


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 ppGalNAc-T2 in different SGC7901 clones of stably transfectedcells, including untransfected SGC7901 cells, SGC7901 cells stably overexpressingppGalNAc-T2 (SGC7901-T2s), SGC7901 cells with downregulated expression of ppGalNAc-T2(SGC7901-T2as) or with empty vector as control. β-actin was used as an internalcontrol for loading. ppGalNAc-T2 expression in different SGC7901 cells was analysed by(A) RT-PCR and (C) western blot analysis. (B and D) Band intensity was quantified usingdensitometry and normalized to β-actin band intensity. 1, Untreated SGC7901cells; 2, control group; 3, SGC7901-T2s group; and 4, SGC7901-T2as group.(*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

f3-ijmm-30-06-1267: Expression levels of ppGalNAc-T2 in different SGC7901 clones of stably transfectedcells, including untransfected SGC7901 cells, SGC7901 cells stably overexpressingppGalNAc-T2 (SGC7901-T2s), SGC7901 cells with downregulated expression of ppGalNAc-T2(SGC7901-T2as) or with empty vector as control. β-actin was used as an internalcontrol for loading. ppGalNAc-T2 expression in different SGC7901 cells was analysed by(A) RT-PCR and (C) western blot analysis. (B and D) Band intensity was quantified usingdensitometry and normalized to β-actin band intensity. 1, Untreated SGC7901cells; 2, control group; 3, SGC7901-T2s group; and 4, SGC7901-T2as group.(*P<0.05, **P>0.05compared to the untreated group).
Mentions: To further explore the role of ppGalNAc-T2 in gastric cancer, SGC7901 cells were used toreconstitute the expression of ppGalNAc-T2 by stable overexpression (SGC7901-T2s) ordownregulation (SGC7901-T2as) of ppGalNAc-T2. Transfection efficiency was measured usingfluorescence microscopy to detect expression of the plasmid-encoded eGFP gene (Fig. 2). Then, the ppGalNAc-T2 mRNA andprotein levels in the SGC7901 cells were measured by RT-PCR and western blot analysis,respectively. When compared with untreated cells, ppGalNAc-T2 transcripts were increasedin the pEGFP-C1-ppGalNAc-T2 sense vector transfected cells (P<0.05) (Fig. 3A and B). Consistent with theRT-PCR results, the expression of the ppGalNAc-T2 protein was clearly increased in thisgroup (P<0.05) (Fig. 3C andD). Furthermore, ppGalNAc-T2 mRNA and protein expression was suppressed in theSGC7901-T2as group when compared to the untreated group (P<0.05), while nodifference was found between the control group and the untreated cells (P>0.05).The above results indicate the successful construction of the ppGalNAc-T2 overexpressionor downregulation cell lines. These stable cell lines can be effectively used to furtherexamine the role of ppGalNAc-T2.

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