Limits...
Caveolin-1 accumulation in the tongue cancer tumor microenvironment is significantly associated with poor prognosis: an in-vivo and in-vitro study.

Vered M, Lehtonen M, Hotakainen L, Pirilä E, Teppo S, Nyberg P, Sormunen R, Zlotogorski-Hurvitz A, Salo T, Dayan D - BMC Cancer (2015)

Bottom Line: RM findings were similar to IM, inferring action of HSC-3 derived factors, and no differences were seen when hypoxia was induced.HSC-3-derived exosomes were loaded with CAV1.Accumulation of CAV1-TME in TSCC had a negative prognostic value.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Pathology and Oral Medicine, School of Dental Medicine, Tel Aviv University, Tel Aviv, 69978, Israel. mvered@post.tau.ac.il.

ABSTRACT

Background: Caveolin-1 (CAV1) may be upregulated by hypoxia and acts in a tumor-dependent manner. We investigated CAV1 in tongue squamous cell carcinoma (TSCC) and its association with clinical outcomes, and studied in vitro possible ways for CAV1 accumulation in the tumor microenvironment (TME).

Methods: TSCC cases (N = 64) were immunohistochemically stained for CAV1. Scores were separately assessed in the tumor and TME and plotted for association with recurrence and survival (univariate analysis with log-rank test). In vitro studies were performed on a 3D myoma organotypic model, a mimicker of TME. Prior to monoculturing HSC-3 tongue cancer cells, the model underwent modifications in oxygenation level (1%O2 hypoxia to upregulate CAV1) and/or in the amount of natural soluble factors [deleted by 14-day rinsing (rinsed myoma, RM), to allow only HSC-3-derived factors to act]. Controls included normoxia (21%O2) and naturally occurring soluble factors (intact myoma, IM). HSC-3 cells were also co-cultured with CaDEC12 cells (fibroblasts exposed to human tongue cancer). CAV1 expression and cellular distribution were examined in different cellular components in hypoxic and rinsed myoma assays. Twist served as a marker for the process of epithelial-mesenchymal transition (EMT). Exosomes isolated from HSC-3 media were investigated for containing CAV1.

Results: Expression of CAV1 in TSCC had a higher score in TME than in the tumor cells and a negative impact on recurrence (p = 0.01) and survival (p = 0.003). Monocultures of HSC-3 revealed expression of CAV1 mainly in the TME-like myoma assay, similar to TSCC. CAV1+, alpha-smooth muscle actin (αSMA) + and Twist + CAF-like cells were observed surrounding the invading HSC-3, possibly reflecting EMT. RM findings were similar to IM, inferring action of HSC-3 derived factors, and no differences were seen when hypoxia was induced. HSC-3-CaDEC12 co-cultures revealed CAV1+, αSMA+ and cytokeratin-negative CAF-like cells, raising the possibility of CaDEC12 cells gaining a CAF phenotype. HSC-3-derived exosomes were loaded with CAV1.

Conclusions: Accumulation of CAV1-TME in TSCC had a negative prognostic value. In vitro studies showed the presence of CAV1 in cancer cells undergoing EMT and in fibroblasts undergoing trans-differentiation to CAFs. CAV1 delivery to the TME involved cancer cell-derived exosomes.

Show MeSH

Related in: MedlinePlus

Caveolin-1 immunostaining patterns in human tongue sections and association with clinical outcomes. A. Staining in the tumor is principally limited to the peripheral cells and is assessed with a score of 1.0 (50% for extent of staining and 2 for staining intensity). In contrast, there is almost no staining of the tumor microenvironment (TME) that was given a score of 0.1 (10% for extent of staining and 1 for staining intensity). B. Most of the tumor cells are strongly positive (a score of 3) and TME is negative (a score of 0). C. Caveolin-1 is moderately positive in the tumor cells (score of 0.9), but it predominates in the TME (score of 3) (A, B, C - bar = 200 μ). D. Kaplan-Meyer analysis shows a significantly negative influence of high CAV-TME score on locoregional recurrence (p = 0.01). E. Kaplan-Meier analysis shows a significantly negative influence of high CAV-TME score on overall survival (p = 0.003).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4318139&req=5

Fig1: Caveolin-1 immunostaining patterns in human tongue sections and association with clinical outcomes. A. Staining in the tumor is principally limited to the peripheral cells and is assessed with a score of 1.0 (50% for extent of staining and 2 for staining intensity). In contrast, there is almost no staining of the tumor microenvironment (TME) that was given a score of 0.1 (10% for extent of staining and 1 for staining intensity). B. Most of the tumor cells are strongly positive (a score of 3) and TME is negative (a score of 0). C. Caveolin-1 is moderately positive in the tumor cells (score of 0.9), but it predominates in the TME (score of 3) (A, B, C - bar = 200 μ). D. Kaplan-Meyer analysis shows a significantly negative influence of high CAV-TME score on locoregional recurrence (p = 0.01). E. Kaplan-Meier analysis shows a significantly negative influence of high CAV-TME score on overall survival (p = 0.003).

Mentions: The present study is the first to investigate the accumulation of CAV1 in cancer cells and TME components in a series of tongue cancer cases and its association with clinical outcome. CAV-SCC was positive in all 64 cases (median score 0.9, range 0.1–3), with only 11 cases being greater than the median and only one case with a score of 3. CAV-TME was positive in 54 cases (median 0.6, range 0–3), with 24 cases being greater than the median and 11 cases having a score of 3. CAV-total had a median of 1.6 (range 0.1–4.5). Figure 1A-C illustrates the CAV1 staining patterns. Univariate analysis demonstrated that only CAV-TME was associated with clinical outcome: a high score had a negative impact on both recurrence (P = 0.01) and survival (P = 0.003) (Figure 1D and E, respectively). Although we detected evidence of CAV1 in the tumor cells in all the examined cases, as reported in previous studies [15-18], we found no significant association with either recurrence or survival. These findings motivated us to perform a series of in vitro studies in which we attempted to determine the mechanisms of CAV1 accumulation within the TME.Figure 1


Caveolin-1 accumulation in the tongue cancer tumor microenvironment is significantly associated with poor prognosis: an in-vivo and in-vitro study.

Vered M, Lehtonen M, Hotakainen L, Pirilä E, Teppo S, Nyberg P, Sormunen R, Zlotogorski-Hurvitz A, Salo T, Dayan D - BMC Cancer (2015)

Caveolin-1 immunostaining patterns in human tongue sections and association with clinical outcomes. A. Staining in the tumor is principally limited to the peripheral cells and is assessed with a score of 1.0 (50% for extent of staining and 2 for staining intensity). In contrast, there is almost no staining of the tumor microenvironment (TME) that was given a score of 0.1 (10% for extent of staining and 1 for staining intensity). B. Most of the tumor cells are strongly positive (a score of 3) and TME is negative (a score of 0). C. Caveolin-1 is moderately positive in the tumor cells (score of 0.9), but it predominates in the TME (score of 3) (A, B, C - bar = 200 μ). D. Kaplan-Meyer analysis shows a significantly negative influence of high CAV-TME score on locoregional recurrence (p = 0.01). E. Kaplan-Meier analysis shows a significantly negative influence of high CAV-TME score on overall survival (p = 0.003).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4318139&req=5

Fig1: Caveolin-1 immunostaining patterns in human tongue sections and association with clinical outcomes. A. Staining in the tumor is principally limited to the peripheral cells and is assessed with a score of 1.0 (50% for extent of staining and 2 for staining intensity). In contrast, there is almost no staining of the tumor microenvironment (TME) that was given a score of 0.1 (10% for extent of staining and 1 for staining intensity). B. Most of the tumor cells are strongly positive (a score of 3) and TME is negative (a score of 0). C. Caveolin-1 is moderately positive in the tumor cells (score of 0.9), but it predominates in the TME (score of 3) (A, B, C - bar = 200 μ). D. Kaplan-Meyer analysis shows a significantly negative influence of high CAV-TME score on locoregional recurrence (p = 0.01). E. Kaplan-Meier analysis shows a significantly negative influence of high CAV-TME score on overall survival (p = 0.003).
Mentions: The present study is the first to investigate the accumulation of CAV1 in cancer cells and TME components in a series of tongue cancer cases and its association with clinical outcome. CAV-SCC was positive in all 64 cases (median score 0.9, range 0.1–3), with only 11 cases being greater than the median and only one case with a score of 3. CAV-TME was positive in 54 cases (median 0.6, range 0–3), with 24 cases being greater than the median and 11 cases having a score of 3. CAV-total had a median of 1.6 (range 0.1–4.5). Figure 1A-C illustrates the CAV1 staining patterns. Univariate analysis demonstrated that only CAV-TME was associated with clinical outcome: a high score had a negative impact on both recurrence (P = 0.01) and survival (P = 0.003) (Figure 1D and E, respectively). Although we detected evidence of CAV1 in the tumor cells in all the examined cases, as reported in previous studies [15-18], we found no significant association with either recurrence or survival. These findings motivated us to perform a series of in vitro studies in which we attempted to determine the mechanisms of CAV1 accumulation within the TME.Figure 1

Bottom Line: RM findings were similar to IM, inferring action of HSC-3 derived factors, and no differences were seen when hypoxia was induced.HSC-3-derived exosomes were loaded with CAV1.Accumulation of CAV1-TME in TSCC had a negative prognostic value.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Pathology and Oral Medicine, School of Dental Medicine, Tel Aviv University, Tel Aviv, 69978, Israel. mvered@post.tau.ac.il.

ABSTRACT

Background: Caveolin-1 (CAV1) may be upregulated by hypoxia and acts in a tumor-dependent manner. We investigated CAV1 in tongue squamous cell carcinoma (TSCC) and its association with clinical outcomes, and studied in vitro possible ways for CAV1 accumulation in the tumor microenvironment (TME).

Methods: TSCC cases (N = 64) were immunohistochemically stained for CAV1. Scores were separately assessed in the tumor and TME and plotted for association with recurrence and survival (univariate analysis with log-rank test). In vitro studies were performed on a 3D myoma organotypic model, a mimicker of TME. Prior to monoculturing HSC-3 tongue cancer cells, the model underwent modifications in oxygenation level (1%O2 hypoxia to upregulate CAV1) and/or in the amount of natural soluble factors [deleted by 14-day rinsing (rinsed myoma, RM), to allow only HSC-3-derived factors to act]. Controls included normoxia (21%O2) and naturally occurring soluble factors (intact myoma, IM). HSC-3 cells were also co-cultured with CaDEC12 cells (fibroblasts exposed to human tongue cancer). CAV1 expression and cellular distribution were examined in different cellular components in hypoxic and rinsed myoma assays. Twist served as a marker for the process of epithelial-mesenchymal transition (EMT). Exosomes isolated from HSC-3 media were investigated for containing CAV1.

Results: Expression of CAV1 in TSCC had a higher score in TME than in the tumor cells and a negative impact on recurrence (p = 0.01) and survival (p = 0.003). Monocultures of HSC-3 revealed expression of CAV1 mainly in the TME-like myoma assay, similar to TSCC. CAV1+, alpha-smooth muscle actin (αSMA) + and Twist + CAF-like cells were observed surrounding the invading HSC-3, possibly reflecting EMT. RM findings were similar to IM, inferring action of HSC-3 derived factors, and no differences were seen when hypoxia was induced. HSC-3-CaDEC12 co-cultures revealed CAV1+, αSMA+ and cytokeratin-negative CAF-like cells, raising the possibility of CaDEC12 cells gaining a CAF phenotype. HSC-3-derived exosomes were loaded with CAV1.

Conclusions: Accumulation of CAV1-TME in TSCC had a negative prognostic value. In vitro studies showed the presence of CAV1 in cancer cells undergoing EMT and in fibroblasts undergoing trans-differentiation to CAFs. CAV1 delivery to the TME involved cancer cell-derived exosomes.

Show MeSH
Related in: MedlinePlus