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Desmocollin‑2 affects the adhesive strength and cytoskeletal arrangement in esophageal squamous cell carcinoma cells.

Fang WK, Liao LD, Zeng FM, Zhang PX, Wu JY, Shen J, Xu LY, Li EM - Mol Med Rep (2014)

Bottom Line: Desmocollin‑2 (DSC2), a transmembrane glycoprotein belonging to the desmosomal cadherin family, has been found to be differentially expressed in several types of cancer and to be involved in tumor progression.The results demonstrated that DSC2 knock down by RNAi caused defects in cell‑cell adhesion and a concomitant reduction in desmosomal protein expression and adherens junction molecule distribution.A decrease in the expression of DSC2 caused an increase in free γ‑catenin levels, thus promoting its recruitment to the adherens junction complex.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Shantou University Medical College, Shantou, Guangdong 515041, P.R. China.

ABSTRACT
Desmocollin‑2 (DSC2), a transmembrane glycoprotein belonging to the desmosomal cadherin family, has been found to be differentially expressed in several types of cancer and to be involved in tumor progression. The tumor metastasis suppressing property of DSC2 in esophageal squamous cell carcinoma (ESCC) has been described, however, its contribution to cell cohesion in ESCC remains to be elucidated. In the present study, using RNA interference (RNAi), the expression of DSC2 was silenced in SHEEC and KYSE510 cells. Hanging drop and fragmentation assays were performed to investigate the role of DSC2 in cell‑cell adhesion. Western blot analysis and confocal microscopy were used to analyze the expression and localization of cell adhesion molecules and cytoskeletal arrangement. The results demonstrated that DSC2 knock down by RNAi caused defects in cell‑cell adhesion and a concomitant reduction in desmosomal protein expression and adherens junction molecule distribution. A decrease in the expression of DSC2 caused an increase in free γ‑catenin levels, thus promoting its recruitment to the adherens junction complex. In addition, the RNAi‑mediated inhibition of DSC2 led to keratin intermediate filament retraction and filamentous‑actin cytoskeleton rearrangement. Taken together, these data support our previous findings and the proposal that DSC2 may be involved in the regulation of the invasive behavior of cells by a mechanism that controls cell‑cell attachment and cytoskeleton rearrangement.

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Related in: MedlinePlus

DSC2 silencing in ESCC cells by siRNAs. (A) RT-qPCR analysis of the expression of DSC2 in ESCC cell lines. The expression levels of DSC2 were normalized to that of β-actin. (B) RT-qPCR analysis of DSC2 silencing by siRNAs. Cells were transfected with DSC2 siRNA or control siRNA. (C) DSC2 silencing in SHEEC and KYSE510 cells was evaluated using western blot analysis. β-actin served as a loading control. (D) Immunofluorescence analysis of DSC2 silencing by siRNAs (magnification, ×400). DSC2, desmocollin-2; ESCC, esophageal squamous cell carcinoma; RT-qPCR, reverse transcription quantitative polymerase chain reaction.
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f1-mmr-10-05-2358: DSC2 silencing in ESCC cells by siRNAs. (A) RT-qPCR analysis of the expression of DSC2 in ESCC cell lines. The expression levels of DSC2 were normalized to that of β-actin. (B) RT-qPCR analysis of DSC2 silencing by siRNAs. Cells were transfected with DSC2 siRNA or control siRNA. (C) DSC2 silencing in SHEEC and KYSE510 cells was evaluated using western blot analysis. β-actin served as a loading control. (D) Immunofluorescence analysis of DSC2 silencing by siRNAs (magnification, ×400). DSC2, desmocollin-2; ESCC, esophageal squamous cell carcinoma; RT-qPCR, reverse transcription quantitative polymerase chain reaction.

Mentions: Initially, the mRNA expression levels of DSC2 were examined in several ESCC cell lines by RT-qPCR. As shown in Fig. 1A, DSC2 mRNA expression was detected in all the cell lines evaluated, with SHEEC and KYSE510 cells containing the highest levels. Therefore, these cell lines were selected for use as the model in the subsequent functional studies.


Desmocollin‑2 affects the adhesive strength and cytoskeletal arrangement in esophageal squamous cell carcinoma cells.

Fang WK, Liao LD, Zeng FM, Zhang PX, Wu JY, Shen J, Xu LY, Li EM - Mol Med Rep (2014)

DSC2 silencing in ESCC cells by siRNAs. (A) RT-qPCR analysis of the expression of DSC2 in ESCC cell lines. The expression levels of DSC2 were normalized to that of β-actin. (B) RT-qPCR analysis of DSC2 silencing by siRNAs. Cells were transfected with DSC2 siRNA or control siRNA. (C) DSC2 silencing in SHEEC and KYSE510 cells was evaluated using western blot analysis. β-actin served as a loading control. (D) Immunofluorescence analysis of DSC2 silencing by siRNAs (magnification, ×400). DSC2, desmocollin-2; ESCC, esophageal squamous cell carcinoma; RT-qPCR, reverse transcription quantitative polymerase chain reaction.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4214350&req=5

f1-mmr-10-05-2358: DSC2 silencing in ESCC cells by siRNAs. (A) RT-qPCR analysis of the expression of DSC2 in ESCC cell lines. The expression levels of DSC2 were normalized to that of β-actin. (B) RT-qPCR analysis of DSC2 silencing by siRNAs. Cells were transfected with DSC2 siRNA or control siRNA. (C) DSC2 silencing in SHEEC and KYSE510 cells was evaluated using western blot analysis. β-actin served as a loading control. (D) Immunofluorescence analysis of DSC2 silencing by siRNAs (magnification, ×400). DSC2, desmocollin-2; ESCC, esophageal squamous cell carcinoma; RT-qPCR, reverse transcription quantitative polymerase chain reaction.
Mentions: Initially, the mRNA expression levels of DSC2 were examined in several ESCC cell lines by RT-qPCR. As shown in Fig. 1A, DSC2 mRNA expression was detected in all the cell lines evaluated, with SHEEC and KYSE510 cells containing the highest levels. Therefore, these cell lines were selected for use as the model in the subsequent functional studies.

Bottom Line: Desmocollin‑2 (DSC2), a transmembrane glycoprotein belonging to the desmosomal cadherin family, has been found to be differentially expressed in several types of cancer and to be involved in tumor progression.The results demonstrated that DSC2 knock down by RNAi caused defects in cell‑cell adhesion and a concomitant reduction in desmosomal protein expression and adherens junction molecule distribution.A decrease in the expression of DSC2 caused an increase in free γ‑catenin levels, thus promoting its recruitment to the adherens junction complex.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Shantou University Medical College, Shantou, Guangdong 515041, P.R. China.

ABSTRACT
Desmocollin‑2 (DSC2), a transmembrane glycoprotein belonging to the desmosomal cadherin family, has been found to be differentially expressed in several types of cancer and to be involved in tumor progression. The tumor metastasis suppressing property of DSC2 in esophageal squamous cell carcinoma (ESCC) has been described, however, its contribution to cell cohesion in ESCC remains to be elucidated. In the present study, using RNA interference (RNAi), the expression of DSC2 was silenced in SHEEC and KYSE510 cells. Hanging drop and fragmentation assays were performed to investigate the role of DSC2 in cell‑cell adhesion. Western blot analysis and confocal microscopy were used to analyze the expression and localization of cell adhesion molecules and cytoskeletal arrangement. The results demonstrated that DSC2 knock down by RNAi caused defects in cell‑cell adhesion and a concomitant reduction in desmosomal protein expression and adherens junction molecule distribution. A decrease in the expression of DSC2 caused an increase in free γ‑catenin levels, thus promoting its recruitment to the adherens junction complex. In addition, the RNAi‑mediated inhibition of DSC2 led to keratin intermediate filament retraction and filamentous‑actin cytoskeleton rearrangement. Taken together, these data support our previous findings and the proposal that DSC2 may be involved in the regulation of the invasive behavior of cells by a mechanism that controls cell‑cell attachment and cytoskeleton rearrangement.

Show MeSH
Related in: MedlinePlus