MicroRNA-33a functions as a bone metastasis suppressor in lung cancer by targeting parathyroid hormone related protein
Bone is a common site of metastasis for lung cancer, and is associated with significant morbidity and a dismal prognosis. MicroRNAs (miRNAs) are increasingly implicated in regulating the progression of malignancies. The efficacy of miR-33a or anti-miR-33a plasmid was assessed by Real-time PCR. Lucif...
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| Vydáno v: | Biochimica et biophysica acta Ročník 1830; číslo 6; s. 3756 - 3766 |
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| Médium: | Journal Article |
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Netherlands
Elsevier B.V
01.06.2013
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| ISSN: | 0304-4165, 0006-3002, 1872-8006 |
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| Abstract | Bone is a common site of metastasis for lung cancer, and is associated with significant morbidity and a dismal prognosis. MicroRNAs (miRNAs) are increasingly implicated in regulating the progression of malignancies.
The efficacy of miR-33a or anti-miR-33a plasmid was assessed by Real-time PCR. Luciferase assays were using One-Glo Luciferase Assay System. Measurement of secreted factors was determined by ELISA kit.
We have found that miR-33a, which is downregulated in lung cancer cells, directly targets PTHrP (parathyroid hormone-related protein), a potent stimulator of osteoclastic bone resorption, leading to decreased osteolytic bone metastasis. We also found that miR-33a levels are inversely correlated with PTHrP expression between human normal bronchial cell line and lung cancer cell lines. The reintroduction of miR-33a reduces the stimulatory effect of A549 on the production of osteoclastogenesis activator RANKL (receptor activator of nuclear factor kappa-B ligand) and M-CSF (macrophage colony-stimulating factor) on osteoblasts, while the expression of PTHrP is decreased in A549 cells. miR-33a overexpression also reduces the inhibitory activity of A549 on the production of OPG (osteoprotegerin), an osteoclastogenesis inhibitor. In addition, miR-33a-mediated PTHrP downregulation results in decreased IL-8 secretion in A549, which contributes to decreased lung cancer-mediated osteoclast differentiation and bone resorption.
These findings have led us to conclude that miR-33a may be a potent tumor suppressor, which inhibits direct and indirect osteoclastogenesis through repression of PTHrP.
miR-33a may even predict a poor prognosis for lung cancer patients.
•miR-33a is downregulated in lung cancer cell lines.•miR-33a regulates the expression of PTHrP.•Overexpression of miR-33a decreases the induction of A549 on the production of RANKL and M-CSF in osteoblasts.•Ectopic expression of miR-33a decreases the stimulatory effect of A549 on osteoclastogenesis.•Knockdown of miR-33a triggers the inductive effect of BEAS-2B on the production of RANKL and M-CSF in osteoblasts. |
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| AbstractList | Bone is a common site of metastasis for lung cancer, and is associated with significant morbidity and a dismal prognosis. MicroRNAs (miRNAs) are increasingly implicated in regulating the progression of malignancies.BACKGROUNDBone is a common site of metastasis for lung cancer, and is associated with significant morbidity and a dismal prognosis. MicroRNAs (miRNAs) are increasingly implicated in regulating the progression of malignancies.The efficacy of miR-33a or anti-miR-33a plasmid was assessed by Real-time PCR. Luciferase assays were using One-Glo Luciferase Assay System. Measurement of secreted factors was determined by ELISA kit.METHODSThe efficacy of miR-33a or anti-miR-33a plasmid was assessed by Real-time PCR. Luciferase assays were using One-Glo Luciferase Assay System. Measurement of secreted factors was determined by ELISA kit.We have found that miR-33a, which is downregulated in lung cancer cells, directly targets PTHrP (parathyroid hormone-related protein), a potent stimulator of osteoclastic bone resorption, leading to decreased osteolytic bone metastasis. We also found that miR-33a levels are inversely correlated with PTHrP expression between human normal bronchial cell line and lung cancer cell lines. The reintroduction of miR-33a reduces the stimulatory effect of A549 on the production of osteoclastogenesis activator RANKL (receptor activator of nuclear factor kappa-B ligand) and M-CSF (macrophage colony-stimulating factor) on osteoblasts, while the expression of PTHrP is decreased in A549 cells. miR-33a overexpression also reduces the inhibitory activity of A549 on the production of OPG (osteoprotegerin), an osteoclastogenesis inhibitor. In addition, miR-33a-mediated PTHrP downregulation results in decreased IL-8 secretion in A549, which contributes to decreased lung cancer-mediated osteoclast differentiation and bone resorption.RESULTSWe have found that miR-33a, which is downregulated in lung cancer cells, directly targets PTHrP (parathyroid hormone-related protein), a potent stimulator of osteoclastic bone resorption, leading to decreased osteolytic bone metastasis. We also found that miR-33a levels are inversely correlated with PTHrP expression between human normal bronchial cell line and lung cancer cell lines. The reintroduction of miR-33a reduces the stimulatory effect of A549 on the production of osteoclastogenesis activator RANKL (receptor activator of nuclear factor kappa-B ligand) and M-CSF (macrophage colony-stimulating factor) on osteoblasts, while the expression of PTHrP is decreased in A549 cells. miR-33a overexpression also reduces the inhibitory activity of A549 on the production of OPG (osteoprotegerin), an osteoclastogenesis inhibitor. In addition, miR-33a-mediated PTHrP downregulation results in decreased IL-8 secretion in A549, which contributes to decreased lung cancer-mediated osteoclast differentiation and bone resorption.These findings have led us to conclude that miR-33a may be a potent tumor suppressor, which inhibits direct and indirect osteoclastogenesis through repression of PTHrP.CONCLUSIONSThese findings have led us to conclude that miR-33a may be a potent tumor suppressor, which inhibits direct and indirect osteoclastogenesis through repression of PTHrP.miR-33a may even predict a poor prognosis for lung cancer patients.GENERAL SIGNIFICANCEmiR-33a may even predict a poor prognosis for lung cancer patients. Bone is a common site of metastasis for lung cancer, and is associated with significant morbidity and a dismal prognosis. MicroRNAs (miRNAs) are increasingly implicated in regulating the progression of malignancies.The efficacy of miR-33a or anti-miR-33a plasmid was assessed by Real-time PCR. Luciferase assays were using One-Glo Luciferase Assay System. Measurement of secreted factors was determined by ELISA kit.We have found that miR-33a, which is downregulated in lung cancer cells, directly targets PTHrP (parathyroid hormone-related protein), a potent stimulator of osteoclastic bone resorption, leading to decreased osteolytic bone metastasis. We also found that miR-33a levels are inversely correlated with PTHrP expression between human normal bronchial cell line and lung cancer cell lines. The reintroduction of miR-33a reduces the stimulatory effect of A549 on the production of osteoclastogenesis activator RANKL (receptor activator of nuclear factor kappa-B ligand) and M-CSF (macrophage colony-stimulating factor) on osteoblasts, while the expression of PTHrP is decreased in A549 cells. miR-33a overexpression also reduces the inhibitory activity of A549 on the production of OPG (osteoprotegerin), an osteoclastogenesis inhibitor. In addition, miR-33a-mediated PTHrP downregulation results in decreased IL-8 secretion in A549, which contributes to decreased lung cancer-mediated osteoclast differentiation and bone resorption.These findings have led us to conclude that miR-33a may be a potent tumor suppressor, which inhibits direct and indirect osteoclastogenesis through repression of PTHrP.miR-33a may even predict a poor prognosis for lung cancer patients. Bone is a common site of metastasis for lung cancer, and is associated with significant morbidity and a dismal prognosis. MicroRNAs (miRNAs) are increasingly implicated in regulating the progression of malignancies. The efficacy of miR-33a or anti-miR-33a plasmid was assessed by Real-time PCR. Luciferase assays were using One-Glo Luciferase Assay System. Measurement of secreted factors was determined by ELISA kit. We have found that miR-33a, which is downregulated in lung cancer cells, directly targets PTHrP (parathyroid hormone-related protein), a potent stimulator of osteoclastic bone resorption, leading to decreased osteolytic bone metastasis. We also found that miR-33a levels are inversely correlated with PTHrP expression between human normal bronchial cell line and lung cancer cell lines. The reintroduction of miR-33a reduces the stimulatory effect of A549 on the production of osteoclastogenesis activator RANKL (receptor activator of nuclear factor kappa-B ligand) and M-CSF (macrophage colony-stimulating factor) on osteoblasts, while the expression of PTHrP is decreased in A549 cells. miR-33a overexpression also reduces the inhibitory activity of A549 on the production of OPG (osteoprotegerin), an osteoclastogenesis inhibitor. In addition, miR-33a-mediated PTHrP downregulation results in decreased IL-8 secretion in A549, which contributes to decreased lung cancer-mediated osteoclast differentiation and bone resorption. These findings have led us to conclude that miR-33a may be a potent tumor suppressor, which inhibits direct and indirect osteoclastogenesis through repression of PTHrP. miR-33a may even predict a poor prognosis for lung cancer patients. •miR-33a is downregulated in lung cancer cell lines.•miR-33a regulates the expression of PTHrP.•Overexpression of miR-33a decreases the induction of A549 on the production of RANKL and M-CSF in osteoblasts.•Ectopic expression of miR-33a decreases the stimulatory effect of A549 on osteoclastogenesis.•Knockdown of miR-33a triggers the inductive effect of BEAS-2B on the production of RANKL and M-CSF in osteoblasts. Bone is a common site of metastasis for lung cancer, and is associated with significant morbidity and a dismal prognosis. MicroRNAs (miRNAs) are increasingly implicated in regulating the progression of malignancies. The efficacy of miR-33a or anti-miR-33a plasmid was assessed by Real-time PCR. Luciferase assays were using One-Glo Luciferase Assay System. Measurement of secreted factors was determined by ELISA kit. We have found that miR-33a, which is downregulated in lung cancer cells, directly targets PTHrP (parathyroid hormone-related protein), a potent stimulator of osteoclastic bone resorption, leading to decreased osteolytic bone metastasis. We also found that miR-33a levels are inversely correlated with PTHrP expression between human normal bronchial cell line and lung cancer cell lines. The reintroduction of miR-33a reduces the stimulatory effect of A549 on the production of osteoclastogenesis activator RANKL (receptor activator of nuclear factor kappa-B ligand) and M-CSF (macrophage colony-stimulating factor) on osteoblasts, while the expression of PTHrP is decreased in A549 cells. miR-33a overexpression also reduces the inhibitory activity of A549 on the production of OPG (osteoprotegerin), an osteoclastogenesis inhibitor. In addition, miR-33a-mediated PTHrP downregulation results in decreased IL-8 secretion in A549, which contributes to decreased lung cancer-mediated osteoclast differentiation and bone resorption. These findings have led us to conclude that miR-33a may be a potent tumor suppressor, which inhibits direct and indirect osteoclastogenesis through repression of PTHrP. miR-33a may even predict a poor prognosis for lung cancer patients. BACKGROUND: Bone is a common site of metastasis for lung cancer, and is associated with significant morbidity and a dismal prognosis. MicroRNAs (miRNAs) are increasingly implicated in regulating the progression of malignancies. METHODS: The efficacy of miR-33a or anti-miR-33a plasmid was assessed by Real-time PCR. Luciferase assays were using One-Glo Luciferase Assay System. Measurement of secreted factors was determined by ELISA kit. RESULTS: We have found that miR-33a, which is downregulated in lung cancer cells, directly targets PTHrP (parathyroid hormone-related protein), a potent stimulator of osteoclastic bone resorption, leading to decreased osteolytic bone metastasis. We also found that miR-33a levels are inversely correlated with PTHrP expression between human normal bronchial cell line and lung cancer cell lines. The reintroduction of miR-33a reduces the stimulatory effect of A549 on the production of osteoclastogenesis activator RANKL (receptor activator of nuclear factor kappa-B ligand) and M-CSF (macrophage colony-stimulating factor) on osteoblasts, while the expression of PTHrP is decreased in A549 cells. miR-33a overexpression also reduces the inhibitory activity of A549 on the production of OPG (osteoprotegerin), an osteoclastogenesis inhibitor. In addition, miR-33a-mediated PTHrP downregulation results in decreased IL-8 secretion in A549, which contributes to decreased lung cancer-mediated osteoclast differentiation and bone resorption. CONCLUSIONS: These findings have led us to conclude that miR-33a may be a potent tumor suppressor, which inhibits direct and indirect osteoclastogenesis through repression of PTHrP. GENERAL SIGNIFICANCE: miR-33a may even predict a poor prognosis for lung cancer patients. |
| Author | Kuo, Po-Lin Huang, Ming-Shyan Hsu, Ya-Ling Liao, Szi-Hui Hung, Jen-Yu |
| Author_xml | – sequence: 1 givenname: Po-Lin surname: Kuo fullname: Kuo, Po-Lin organization: Institute of Clinical Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan – sequence: 2 givenname: Szi-Hui surname: Liao fullname: Liao, Szi-Hui organization: Graduate Institute of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan – sequence: 3 givenname: Jen-Yu surname: Hung fullname: Hung, Jen-Yu organization: Cancer Center, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan – sequence: 4 givenname: Ming-Shyan surname: Huang fullname: Huang, Ming-Shyan organization: Cancer Center, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan – sequence: 5 givenname: Ya-Ling surname: Hsu fullname: Hsu, Ya-Ling email: hsuyl326@gmail.com organization: Graduate Institute of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/23458685$$D View this record in MEDLINE/PubMed |
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| Keywords | Osteoclastogenesis RANKL M-CSF mir-33a Lung cancer OPG Real-time qRT-PCR PTHrP |
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| Snippet | Bone is a common site of metastasis for lung cancer, and is associated with significant morbidity and a dismal prognosis. MicroRNAs (miRNAs) are increasingly... BACKGROUND: Bone is a common site of metastasis for lung cancer, and is associated with significant morbidity and a dismal prognosis. MicroRNAs (miRNAs) are... |
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| SubjectTerms | Bone Neoplasms - genetics Bone Neoplasms - metabolism Bone Neoplasms - pathology Bone Neoplasms - secondary bone resorption Bone Resorption - genetics Bone Resorption - metabolism Bone Resorption - pathology bones Cell Differentiation - genetics Cell Line, Tumor Down-Regulation - genetics enzyme-linked immunosorbent assay Gene Expression Regulation, Neoplastic Genes, Tumor Suppressor Humans interleukin-8 Interleukin-8 - biosynthesis Interleukin-8 - genetics luciferase Lung cancer lung neoplasms Lung Neoplasms - genetics Lung Neoplasms - metabolism Lung Neoplasms - pathology macrophage colony-stimulating factor metastasis microRNA MicroRNAs - biosynthesis MicroRNAs - genetics mir-33a morbidity neoplasm cells Neoplasm Metastasis Neoplasm Proteins - biosynthesis Neoplasm Proteins - genetics OPG osteoblasts Osteoclastogenesis osteoclasts Osteoclasts - metabolism Osteoclasts - pathology Osteoprotegerin - biosynthesis Osteoprotegerin - genetics parathyroid hormone Parathyroid Hormone-Related Protein - biosynthesis Parathyroid Hormone-Related Protein - genetics patients plasmids prognosis PTHrP quantitative polymerase chain reaction RANK Ligand - biosynthesis RANK Ligand - genetics RNA, Neoplasm - biosynthesis RNA, Neoplasm - genetics secretion |
| Title | MicroRNA-33a functions as a bone metastasis suppressor in lung cancer by targeting parathyroid hormone related protein |
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