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Inhibition of Akt activity induces the mesenchymal-to-epithelial reverting transition with restoring E-cadherin expression in KB and KOSCC-25B oral squamous cell carcinoma cells.

Hong KO, Kim JH, Hong JS, Yoon HJ, Lee JI, Hong SP, Hong SD - J. Exp. Clin. Cancer Res. (2009)

Bottom Line: Inhibition of Akt activity by PIA decreased NF-kappaB signaling, but did not affect phosphorylation of ERK, JNK, and p38 in KB and KOSCC-25B cells.In contrast, inhibition of Akt activity by PIA did not induce any changes in SIP-1/ZEB-2 expression.All of these findings suggest that Akt inhibition could induce the MErT through decreased NF-kappaB signaling and downregulation of Snail and Twist in OSCC cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Oral Pathology, School of Dentistry and Dental Research Institute, Seoul National University, Seoul, Korea. hongko95@hanmail.net

ABSTRACT

Background: The Akt/PKB family of kinases is frequently activated in human cancers, including oral squamous cell carcinoma (OSCC). Akt-induced epithelial-to-mesenchymal transition (EMT) involves downregulation of E-cadherin, which appears to result from upregulation of the transcription repressor Snail. Recently, it was proposed that carcinoma cells, especially in metastatic sites, could acquire the mesenchymal-to-epithelial reverting transition (MErT) in order to adapt the microenvironments and re-expression of E-cadherin be a critical indicator of MErT. However, the precise mechanism and biologic or clinical importance of the MErT in cancers have been little known. This study aimed to investigate whether Akt inhibition would restore the expression of E-cadherin and beta-catenin, reduce that of Vimentin, and induce the MErT in OSCC cells with low or negative expression of E-cadherin. We also investigate whether inhibition of Akt activity would affect the E-cadherin repressors and signaling molecules like NF-kappaB, ERK, and p38.

Methods: We screened several OSCC cell lines in order to select suitable cell line models for inducing MErT, using immunoblotting and methylation specific-PCR. We examined whether Akt inhibitor phosphatidylinositol ether lipid analogues (PIA) treatment would restore the expression of E-cadherin and beta-catenin, reduce that of Vimentin, and induce the MErT in KB and KOSCC-25B cells using RT-PCR, immunoblotting, immunofluorescence analysis, and in vitro migration assay. We also investigated whether inhibition of Akt activity would affect the E-cadherin repressors, including Snail, Twist, and SIP-1/ZEB-2 and signaling molecules like NF-kappaB, ERK, JNK, and p38 using RT-PCR, immunoblotting, and immunofluorescence analysis.

Results: Of the 7 OSCC cell lines, KB and KOSCC-25B showed constitutively activated phosphorylated Akt and low or negative expression of E-cadherin. Inhibition of Akt activity by PIA decreased NF-kappaB signaling, but did not affect phosphorylation of ERK, JNK, and p38 in KB and KOSCC-25B cells. Akt inhibition led to downregulation of Snail and Twist expression. In contrast, inhibition of Akt activity by PIA did not induce any changes in SIP-1/ZEB-2 expression. PIA treatment induced the expression of E-cadherin and beta-catenin, reduce that of Vimentin, restored their epithelial morphology of a polygonal shape, and reduced tumor cell migration in KB and KOSCC-25B cells, which was the corresponding feature of MErT.

Conclusion: All of these findings suggest that Akt inhibition could induce the MErT through decreased NF-kappaB signaling and downregulation of Snail and Twist in OSCC cells. A strategy involving Akt inhibition might be a useful therapeutic tool in controlling cancer dissemination and metastasis in oral cancer patients.

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Related in: MedlinePlus

Reduced migratory ability due to Akt inhibition. Photomicrography of control (A) and PIA-treated (B) KOSCC-25B groups in the in vitro migration assay. (C) The numbers of KB and KOSCC-25B cells from the PIA-treated group that migrated through the filter were only 61.1% and 56.4% of that in control cells (P < 0.05), respectively.
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Figure 6: Reduced migratory ability due to Akt inhibition. Photomicrography of control (A) and PIA-treated (B) KOSCC-25B groups in the in vitro migration assay. (C) The numbers of KB and KOSCC-25B cells from the PIA-treated group that migrated through the filter were only 61.1% and 56.4% of that in control cells (P < 0.05), respectively.

Mentions: In order to examine whether inhibition of Akt activity could affect cell motility, we performed an in vitro migration assay. The numbers of KB and KOSCC-25B cells from the PIA-treated group that migrated through the filter were only 61.1% and 56.4% of that in control cells (P < 0.05; Fig. 6), respectively.


Inhibition of Akt activity induces the mesenchymal-to-epithelial reverting transition with restoring E-cadherin expression in KB and KOSCC-25B oral squamous cell carcinoma cells.

Hong KO, Kim JH, Hong JS, Yoon HJ, Lee JI, Hong SP, Hong SD - J. Exp. Clin. Cancer Res. (2009)

Reduced migratory ability due to Akt inhibition. Photomicrography of control (A) and PIA-treated (B) KOSCC-25B groups in the in vitro migration assay. (C) The numbers of KB and KOSCC-25B cells from the PIA-treated group that migrated through the filter were only 61.1% and 56.4% of that in control cells (P < 0.05), respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2651854&req=5

Figure 6: Reduced migratory ability due to Akt inhibition. Photomicrography of control (A) and PIA-treated (B) KOSCC-25B groups in the in vitro migration assay. (C) The numbers of KB and KOSCC-25B cells from the PIA-treated group that migrated through the filter were only 61.1% and 56.4% of that in control cells (P < 0.05), respectively.
Mentions: In order to examine whether inhibition of Akt activity could affect cell motility, we performed an in vitro migration assay. The numbers of KB and KOSCC-25B cells from the PIA-treated group that migrated through the filter were only 61.1% and 56.4% of that in control cells (P < 0.05; Fig. 6), respectively.

Bottom Line: Inhibition of Akt activity by PIA decreased NF-kappaB signaling, but did not affect phosphorylation of ERK, JNK, and p38 in KB and KOSCC-25B cells.In contrast, inhibition of Akt activity by PIA did not induce any changes in SIP-1/ZEB-2 expression.All of these findings suggest that Akt inhibition could induce the MErT through decreased NF-kappaB signaling and downregulation of Snail and Twist in OSCC cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Oral Pathology, School of Dentistry and Dental Research Institute, Seoul National University, Seoul, Korea. hongko95@hanmail.net

ABSTRACT

Background: The Akt/PKB family of kinases is frequently activated in human cancers, including oral squamous cell carcinoma (OSCC). Akt-induced epithelial-to-mesenchymal transition (EMT) involves downregulation of E-cadherin, which appears to result from upregulation of the transcription repressor Snail. Recently, it was proposed that carcinoma cells, especially in metastatic sites, could acquire the mesenchymal-to-epithelial reverting transition (MErT) in order to adapt the microenvironments and re-expression of E-cadherin be a critical indicator of MErT. However, the precise mechanism and biologic or clinical importance of the MErT in cancers have been little known. This study aimed to investigate whether Akt inhibition would restore the expression of E-cadherin and beta-catenin, reduce that of Vimentin, and induce the MErT in OSCC cells with low or negative expression of E-cadherin. We also investigate whether inhibition of Akt activity would affect the E-cadherin repressors and signaling molecules like NF-kappaB, ERK, and p38.

Methods: We screened several OSCC cell lines in order to select suitable cell line models for inducing MErT, using immunoblotting and methylation specific-PCR. We examined whether Akt inhibitor phosphatidylinositol ether lipid analogues (PIA) treatment would restore the expression of E-cadherin and beta-catenin, reduce that of Vimentin, and induce the MErT in KB and KOSCC-25B cells using RT-PCR, immunoblotting, immunofluorescence analysis, and in vitro migration assay. We also investigated whether inhibition of Akt activity would affect the E-cadherin repressors, including Snail, Twist, and SIP-1/ZEB-2 and signaling molecules like NF-kappaB, ERK, JNK, and p38 using RT-PCR, immunoblotting, and immunofluorescence analysis.

Results: Of the 7 OSCC cell lines, KB and KOSCC-25B showed constitutively activated phosphorylated Akt and low or negative expression of E-cadherin. Inhibition of Akt activity by PIA decreased NF-kappaB signaling, but did not affect phosphorylation of ERK, JNK, and p38 in KB and KOSCC-25B cells. Akt inhibition led to downregulation of Snail and Twist expression. In contrast, inhibition of Akt activity by PIA did not induce any changes in SIP-1/ZEB-2 expression. PIA treatment induced the expression of E-cadherin and beta-catenin, reduce that of Vimentin, restored their epithelial morphology of a polygonal shape, and reduced tumor cell migration in KB and KOSCC-25B cells, which was the corresponding feature of MErT.

Conclusion: All of these findings suggest that Akt inhibition could induce the MErT through decreased NF-kappaB signaling and downregulation of Snail and Twist in OSCC cells. A strategy involving Akt inhibition might be a useful therapeutic tool in controlling cancer dissemination and metastasis in oral cancer patients.

Show MeSH
Related in: MedlinePlus