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A Novel Function of CD82/KAI1 in Sialyl Lewis Antigen-Mediated Adhesion of Cancer Cells: Evidence for an Anti-Metastasis Effect by Down-Regulation of Sialyl Lewis Antigens.

Yoshihama N, Yamaguchi K, Chigita S, Mine M, Abe M, Ishii K, Kobayashi Y, Akimoto N, Mori Y, Sugiura T - PLoS ONE (2015)

Bottom Line: In addition, cells over-expressing CD82 exhibited reduced expression of sLea/x compared to CD82-negative wild-type cells.Significant down-regulation of ST3 β-galactoside α-2, 3-sialyltransferase 4 (ST3GAL4) was detected by cDNA microarray, real-time PCR, and western blotting analyses.We concluded that CD82 decreases sLea/x expression via the down-regulation of ST3GAL4 expression and thereby reduces the adhesion of cancer cells to blood vessels, which results in inhibition of metastasis.

View Article: PubMed Central - PubMed

Affiliation: Section of Oral and Maxillofacial Surgery, Division of Maxillofacial Diagnostic and Surgical Sciences, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan.

ABSTRACT
We have recently elucidated a novel function for CD82 in E-cadherin-mediated homocellular adhesion; due to this function, it can inhibit cancer cell dissociation from the primary cancer nest and limit metastasis. However, the effect of CD82 on selectin ligand-mediated heterocellular adhesion has not yet been elucidated. In this study, we focused on the effects of the metastasis suppressor CD82/KAI1 on heterocellular adhesion of cancer cells to the endothelium of blood vessels in order to further elucidate the function of tetraspanins. The over-expression of CD82 in cancer cells led to the inhibition of experimentally induced lung metastases in mice and significantly inhibited the adhesion of these cells to human umbilical vein epithelial cells (HUVECs) in vitro. Pre-treatment of the cells with function-perturbing antibodies against sLea/x significantly inhibited the adhesion of CD82-negative cells to HUVECs. In addition, cells over-expressing CD82 exhibited reduced expression of sLea/x compared to CD82-negative wild-type cells. Significant down-regulation of ST3 β-galactoside α-2, 3-sialyltransferase 4 (ST3GAL4) was detected by cDNA microarray, real-time PCR, and western blotting analyses. Knockdown of ST3GAL4 on CD82-negative wild-type cells inhibited expression of sLex and reduced cell adhesion to HUVECs. We concluded that CD82 decreases sLea/x expression via the down-regulation of ST3GAL4 expression and thereby reduces the adhesion of cancer cells to blood vessels, which results in inhibition of metastasis.

No MeSH data available.


Related in: MedlinePlus

Effects of CD82 on the expression of sialyl Lewis antigens.Whole cell lysates (200 μg) of h1299 cells (h1299/zeo, zeo; h1299/CD82, CD82; h1299/CD82-sh.control, sh.cont; h1299/CD82-sh.CD82, sh.CD82) were resolved by 7.5% SDS-PAGE and analysed by immunoblotting with anti-sLea (A) or anti-sLex (B) antibodies. The same blots were stripped and re-probed for β-actin as a loading control. Experiments were repeated in triplicate, and the most representative data are shown.
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pone.0124743.g003: Effects of CD82 on the expression of sialyl Lewis antigens.Whole cell lysates (200 μg) of h1299 cells (h1299/zeo, zeo; h1299/CD82, CD82; h1299/CD82-sh.control, sh.cont; h1299/CD82-sh.CD82, sh.CD82) were resolved by 7.5% SDS-PAGE and analysed by immunoblotting with anti-sLea (A) or anti-sLex (B) antibodies. The same blots were stripped and re-probed for β-actin as a loading control. Experiments were repeated in triplicate, and the most representative data are shown.

Mentions: The expression of sialyl Lewis antigens on h1299 cells was analysed by immunoblotting (Fig 3). sLea was detected as bands corresponding to approximately 97, 125 and 200 kDa, and its expression level was 5–8-fold higher in h1299/zeo cells than in h1299/CD82 cells (Fig 3A). Similarly, the expression of sLex proteins was detected at approximately 90, 125, and 200 kDa, and was 5–8-fold higher in h1299/zeo cells than in h1299/CD82 cells (Fig 3B). These findings were in accordance with the results of the HUVEC adhesion assay.


A Novel Function of CD82/KAI1 in Sialyl Lewis Antigen-Mediated Adhesion of Cancer Cells: Evidence for an Anti-Metastasis Effect by Down-Regulation of Sialyl Lewis Antigens.

Yoshihama N, Yamaguchi K, Chigita S, Mine M, Abe M, Ishii K, Kobayashi Y, Akimoto N, Mori Y, Sugiura T - PLoS ONE (2015)

Effects of CD82 on the expression of sialyl Lewis antigens.Whole cell lysates (200 μg) of h1299 cells (h1299/zeo, zeo; h1299/CD82, CD82; h1299/CD82-sh.control, sh.cont; h1299/CD82-sh.CD82, sh.CD82) were resolved by 7.5% SDS-PAGE and analysed by immunoblotting with anti-sLea (A) or anti-sLex (B) antibodies. The same blots were stripped and re-probed for β-actin as a loading control. Experiments were repeated in triplicate, and the most representative data are shown.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4414583&req=5

pone.0124743.g003: Effects of CD82 on the expression of sialyl Lewis antigens.Whole cell lysates (200 μg) of h1299 cells (h1299/zeo, zeo; h1299/CD82, CD82; h1299/CD82-sh.control, sh.cont; h1299/CD82-sh.CD82, sh.CD82) were resolved by 7.5% SDS-PAGE and analysed by immunoblotting with anti-sLea (A) or anti-sLex (B) antibodies. The same blots were stripped and re-probed for β-actin as a loading control. Experiments were repeated in triplicate, and the most representative data are shown.
Mentions: The expression of sialyl Lewis antigens on h1299 cells was analysed by immunoblotting (Fig 3). sLea was detected as bands corresponding to approximately 97, 125 and 200 kDa, and its expression level was 5–8-fold higher in h1299/zeo cells than in h1299/CD82 cells (Fig 3A). Similarly, the expression of sLex proteins was detected at approximately 90, 125, and 200 kDa, and was 5–8-fold higher in h1299/zeo cells than in h1299/CD82 cells (Fig 3B). These findings were in accordance with the results of the HUVEC adhesion assay.

Bottom Line: In addition, cells over-expressing CD82 exhibited reduced expression of sLea/x compared to CD82-negative wild-type cells.Significant down-regulation of ST3 β-galactoside α-2, 3-sialyltransferase 4 (ST3GAL4) was detected by cDNA microarray, real-time PCR, and western blotting analyses.We concluded that CD82 decreases sLea/x expression via the down-regulation of ST3GAL4 expression and thereby reduces the adhesion of cancer cells to blood vessels, which results in inhibition of metastasis.

View Article: PubMed Central - PubMed

Affiliation: Section of Oral and Maxillofacial Surgery, Division of Maxillofacial Diagnostic and Surgical Sciences, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan.

ABSTRACT
We have recently elucidated a novel function for CD82 in E-cadherin-mediated homocellular adhesion; due to this function, it can inhibit cancer cell dissociation from the primary cancer nest and limit metastasis. However, the effect of CD82 on selectin ligand-mediated heterocellular adhesion has not yet been elucidated. In this study, we focused on the effects of the metastasis suppressor CD82/KAI1 on heterocellular adhesion of cancer cells to the endothelium of blood vessels in order to further elucidate the function of tetraspanins. The over-expression of CD82 in cancer cells led to the inhibition of experimentally induced lung metastases in mice and significantly inhibited the adhesion of these cells to human umbilical vein epithelial cells (HUVECs) in vitro. Pre-treatment of the cells with function-perturbing antibodies against sLea/x significantly inhibited the adhesion of CD82-negative cells to HUVECs. In addition, cells over-expressing CD82 exhibited reduced expression of sLea/x compared to CD82-negative wild-type cells. Significant down-regulation of ST3 β-galactoside α-2, 3-sialyltransferase 4 (ST3GAL4) was detected by cDNA microarray, real-time PCR, and western blotting analyses. Knockdown of ST3GAL4 on CD82-negative wild-type cells inhibited expression of sLex and reduced cell adhesion to HUVECs. We concluded that CD82 decreases sLea/x expression via the down-regulation of ST3GAL4 expression and thereby reduces the adhesion of cancer cells to blood vessels, which results in inhibition of metastasis.

No MeSH data available.


Related in: MedlinePlus