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miR-126-3p Inhibits Thyroid Cancer Cell Growth and Metastasis, and Is Associated with Aggressive Thyroid Cancer.

Xiong Y, Kotian S, Zeiger MA, Zhang L, Kebebew E - PLoS ONE (2015)

Bottom Line: We found that miR-126-3p expression was significantly lower in larger tumors, in tumor samples with extrathyroidal invasion, and in higher risk group thyroid cancer in 496 papillary thyroid cancer samples from The Cancer Genome Atlas study cohort.In an independent sample set, lower miR-126-3p expression was observed in follicular thyroid cancers (which have capsular and angioinvasion) as compared to follicular adenomas.Of these 14 genes, SLC7A5 and ADAM9 were confirmed to be inhibited by miR-126-3p overexpression and to be direct targets of miR-136-3p.

View Article: PubMed Central - PubMed

Affiliation: Endocrine Oncology Branch, Center for Cancer Research, National Cancer Institute, Bethesda, Maryland, United States of America.

ABSTRACT

Background: Previous studies have shown that microRNAs are dysregulated in thyroid cancer and play important roles in the post-transcriptional regulation of target oncogenes and/or tumor suppressor genes.

Methodology/principal findings: We studied the function of miR-126-3p in thyroid cancer cells, and as a marker of disease aggressiveness. We found that miR-126-3p expression was significantly lower in larger tumors, in tumor samples with extrathyroidal invasion, and in higher risk group thyroid cancer in 496 papillary thyroid cancer samples from The Cancer Genome Atlas study cohort. In an independent sample set, lower miR-126-3p expression was observed in follicular thyroid cancers (which have capsular and angioinvasion) as compared to follicular adenomas. Mechanistically, ectopic overexpression of miR-126-3p significantly inhibited thyroid cancer cell proliferation, in vitro (p<0.01) and in vivo (p<0.01), colony formation (p<0.01), tumor spheroid formation (p<0.05), cellular migration (p<0.05), VEGF secretion and endothelial tube formation, and lung metastasis in vivo. We found 14 predicted target genes, which were significantly altered upon miR-126-3p transfection in thyroid cancer cells, and which are involved in cancer biology. Of these 14 genes, SLC7A5 and ADAM9 were confirmed to be inhibited by miR-126-3p overexpression and to be direct targets of miR-136-3p.

Conclusions/significance: To our knowledge, this is the first study to demonstrate that miR-126-3p has a tumor-suppressive function in thyroid cancer cells, and is associated with aggressive disease phenotype.

No MeSH data available.


Related in: MedlinePlus

miR-126-3p reduces VEGF secretion and endothelial tube formation.(A) miR-126-3p overexpression significantly reduced secreted VEGF levels in TPC-1 and FTC-133 cell lines but not XTC-1 cells. * p < 0.5. VEGF levels were also normalized to total protein. Culture media was harvested from TPC-1, FTC-133 and XTC-1 cells transfected with miR-NC and miR-126-3p after 72 hours of transfection. (B) Expression of VEGF protein in lung tumor metastases was decreased in mice injected with FTC-133 Luc cells transfected with miR-126-3p as compared to miR-NC. (C) Endothelial cell tube formation is decreased with miR-126-3p overexpression. All experiments were repeated at least three times.
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pone.0130496.g006: miR-126-3p reduces VEGF secretion and endothelial tube formation.(A) miR-126-3p overexpression significantly reduced secreted VEGF levels in TPC-1 and FTC-133 cell lines but not XTC-1 cells. * p < 0.5. VEGF levels were also normalized to total protein. Culture media was harvested from TPC-1, FTC-133 and XTC-1 cells transfected with miR-NC and miR-126-3p after 72 hours of transfection. (B) Expression of VEGF protein in lung tumor metastases was decreased in mice injected with FTC-133 Luc cells transfected with miR-126-3p as compared to miR-NC. (C) Endothelial cell tube formation is decreased with miR-126-3p overexpression. All experiments were repeated at least three times.

Mentions: Given that we found miR-126-3p expression is lower in localized follicular thyroid cancer with capsular and vascular invasion, and miR-126-3p has been reported to regulate angiogenesis and target VEGF, we next determined whether miR-126-3p regulates angiogenesis in thyroid cancer cells [17–20]. We found that miR-126-3p overexpression significantly decreased VEGF secretion in two of three thyroid cancer cells in vitro (Fig 6A). One of the principal determinants for successful tumor growth is the ability to recruit new blood vessels. Thus, we analyzed the VEGF protein expression level in the lung metastatic tumors from the in vivo studies and endothelial tube formation by using the HUVEC assay in vitro. We found that miR-126-3p overexpression decreased VEGF protein expression in metastatic tumor xenografts cells and reduced endothelial cell tube formation (Fig 6B and 6C). These findings are consistent with our results of reduced miR-126-3p expression in localized follicular thyroid cancer with angioinvasion.


miR-126-3p Inhibits Thyroid Cancer Cell Growth and Metastasis, and Is Associated with Aggressive Thyroid Cancer.

Xiong Y, Kotian S, Zeiger MA, Zhang L, Kebebew E - PLoS ONE (2015)

miR-126-3p reduces VEGF secretion and endothelial tube formation.(A) miR-126-3p overexpression significantly reduced secreted VEGF levels in TPC-1 and FTC-133 cell lines but not XTC-1 cells. * p < 0.5. VEGF levels were also normalized to total protein. Culture media was harvested from TPC-1, FTC-133 and XTC-1 cells transfected with miR-NC and miR-126-3p after 72 hours of transfection. (B) Expression of VEGF protein in lung tumor metastases was decreased in mice injected with FTC-133 Luc cells transfected with miR-126-3p as compared to miR-NC. (C) Endothelial cell tube formation is decreased with miR-126-3p overexpression. All experiments were repeated at least three times.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4526518&req=5

pone.0130496.g006: miR-126-3p reduces VEGF secretion and endothelial tube formation.(A) miR-126-3p overexpression significantly reduced secreted VEGF levels in TPC-1 and FTC-133 cell lines but not XTC-1 cells. * p < 0.5. VEGF levels were also normalized to total protein. Culture media was harvested from TPC-1, FTC-133 and XTC-1 cells transfected with miR-NC and miR-126-3p after 72 hours of transfection. (B) Expression of VEGF protein in lung tumor metastases was decreased in mice injected with FTC-133 Luc cells transfected with miR-126-3p as compared to miR-NC. (C) Endothelial cell tube formation is decreased with miR-126-3p overexpression. All experiments were repeated at least three times.
Mentions: Given that we found miR-126-3p expression is lower in localized follicular thyroid cancer with capsular and vascular invasion, and miR-126-3p has been reported to regulate angiogenesis and target VEGF, we next determined whether miR-126-3p regulates angiogenesis in thyroid cancer cells [17–20]. We found that miR-126-3p overexpression significantly decreased VEGF secretion in two of three thyroid cancer cells in vitro (Fig 6A). One of the principal determinants for successful tumor growth is the ability to recruit new blood vessels. Thus, we analyzed the VEGF protein expression level in the lung metastatic tumors from the in vivo studies and endothelial tube formation by using the HUVEC assay in vitro. We found that miR-126-3p overexpression decreased VEGF protein expression in metastatic tumor xenografts cells and reduced endothelial cell tube formation (Fig 6B and 6C). These findings are consistent with our results of reduced miR-126-3p expression in localized follicular thyroid cancer with angioinvasion.

Bottom Line: We found that miR-126-3p expression was significantly lower in larger tumors, in tumor samples with extrathyroidal invasion, and in higher risk group thyroid cancer in 496 papillary thyroid cancer samples from The Cancer Genome Atlas study cohort.In an independent sample set, lower miR-126-3p expression was observed in follicular thyroid cancers (which have capsular and angioinvasion) as compared to follicular adenomas.Of these 14 genes, SLC7A5 and ADAM9 were confirmed to be inhibited by miR-126-3p overexpression and to be direct targets of miR-136-3p.

View Article: PubMed Central - PubMed

Affiliation: Endocrine Oncology Branch, Center for Cancer Research, National Cancer Institute, Bethesda, Maryland, United States of America.

ABSTRACT

Background: Previous studies have shown that microRNAs are dysregulated in thyroid cancer and play important roles in the post-transcriptional regulation of target oncogenes and/or tumor suppressor genes.

Methodology/principal findings: We studied the function of miR-126-3p in thyroid cancer cells, and as a marker of disease aggressiveness. We found that miR-126-3p expression was significantly lower in larger tumors, in tumor samples with extrathyroidal invasion, and in higher risk group thyroid cancer in 496 papillary thyroid cancer samples from The Cancer Genome Atlas study cohort. In an independent sample set, lower miR-126-3p expression was observed in follicular thyroid cancers (which have capsular and angioinvasion) as compared to follicular adenomas. Mechanistically, ectopic overexpression of miR-126-3p significantly inhibited thyroid cancer cell proliferation, in vitro (p<0.01) and in vivo (p<0.01), colony formation (p<0.01), tumor spheroid formation (p<0.05), cellular migration (p<0.05), VEGF secretion and endothelial tube formation, and lung metastasis in vivo. We found 14 predicted target genes, which were significantly altered upon miR-126-3p transfection in thyroid cancer cells, and which are involved in cancer biology. Of these 14 genes, SLC7A5 and ADAM9 were confirmed to be inhibited by miR-126-3p overexpression and to be direct targets of miR-136-3p.

Conclusions/significance: To our knowledge, this is the first study to demonstrate that miR-126-3p has a tumor-suppressive function in thyroid cancer cells, and is associated with aggressive disease phenotype.

No MeSH data available.


Related in: MedlinePlus