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MicroRNA-144 suppresses osteosarcoma growth and metastasis by targeting ROCK1 and ROCK2.

Wang W, Zhou X, Wei M - Oncotarget (2015)

Bottom Line: Low-level expression of miR-144 was significantly associated with distant metastasis and poor prognosis.Furthermore, we identified Rho-associated kinases 1 and 2 (ROCK1 and ROCK2) as direct targets for miR-144 binding, resulting in suppression of their expression.In clinical OS specimens, ROCK1 and ROCK2 levels were elevated, relative to that in paired normal bone tissues, and inversely correlated with miR-144 expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedic Surgery, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200127, China.

ABSTRACT
Osteosarcoma (OS) is the most common primary tumor of bone. MicroRNAs (miRNAs) are a class of endogenously expressed small non-coding RNAs that are strongly implicated in cancerous processes. However, our current understanding of the biological role of miRNAs in OS remains incomplete. In the present study, miR-144 was markedly downregulated in OS cell lines and clinical specimens. Low-level expression of miR-144 was significantly associated with distant metastasis and poor prognosis. Functional studies demonstrated that ectopic expression of miR-144 suppresses tumor cell proliferation and metastasis in vitro as well as in vivo. Furthermore, we identified Rho-associated kinases 1 and 2 (ROCK1 and ROCK2) as direct targets for miR-144 binding, resulting in suppression of their expression. Exogenous expression of ROCK1 or ROCK2 in 143B-miR-144 cells partially restored miR-144-inhibited cell proliferation and invasion. In clinical OS specimens, ROCK1 and ROCK2 levels were elevated, relative to that in paired normal bone tissues, and inversely correlated with miR-144 expression. Taken together, miR-144 suppresses OS progression by directly downregulating ROCK1 and ROCK2 expression, and may be a promising therapeutic target for OS.

No MeSH data available.


Related in: MedlinePlus

miR-144 suppresses OS cell proliferation in vitroA. 143B cells were transiently transfected with miR-144 mimics or NC mimics, and cell viability determined with the CCK-8 assay. B. FACS analysis of 143B cells was performed after transfection with miR-144 mimics or NC mimics. C. The apoptosis cells were stained with Annexin V and PI, and analyzed using FACS. D. Representative photographs and quantitative analysis of plate colony formation of 143B cells stably expressing miR-144. Silencing of miR-144 in Saos-2 cells significantly promoted proliferation E. and cell cycle G1/S transition F. *P < 0.05, **P < 0.01.
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Figure 2: miR-144 suppresses OS cell proliferation in vitroA. 143B cells were transiently transfected with miR-144 mimics or NC mimics, and cell viability determined with the CCK-8 assay. B. FACS analysis of 143B cells was performed after transfection with miR-144 mimics or NC mimics. C. The apoptosis cells were stained with Annexin V and PI, and analyzed using FACS. D. Representative photographs and quantitative analysis of plate colony formation of 143B cells stably expressing miR-144. Silencing of miR-144 in Saos-2 cells significantly promoted proliferation E. and cell cycle G1/S transition F. *P < 0.05, **P < 0.01.

Mentions: To explore the potential role of miR-144 in OS pathogenesis, 143B cells with high metastatic potential and low endogenous miR-144 expression, were transfected with miR-144 mimics, and miR-144 expression assayed using qRT-PCR (Supplementary Fig. 1A). Data from the cell viability assay showed that upregulation of miR-144 significantly suppresses the proliferation of 143B cells, compared with miR-NC-transfected cells (Fig. 2A). Flow cytometry analysis revealed that miR-144 overexpression leads to an increased percentage of cells in the G1 phase, alone with a decrease in S-phase cells (Fig. 2B), suggesting that this miRNA induces G1/S arrest. Moreover, the rate of apoptosis was significantly higher in 143B cells overexpressing miR-144 (Fig. 2C). The lentivirus system expressing miR-144-GFP was additionally applied to generate 143B-miR-144 cells stably expressing miR-144 (Supplementary Fig. 1B). As expected, the colony formation rate of these cells was significantly decreased (Fig. 2D).


MicroRNA-144 suppresses osteosarcoma growth and metastasis by targeting ROCK1 and ROCK2.

Wang W, Zhou X, Wei M - Oncotarget (2015)

miR-144 suppresses OS cell proliferation in vitroA. 143B cells were transiently transfected with miR-144 mimics or NC mimics, and cell viability determined with the CCK-8 assay. B. FACS analysis of 143B cells was performed after transfection with miR-144 mimics or NC mimics. C. The apoptosis cells were stained with Annexin V and PI, and analyzed using FACS. D. Representative photographs and quantitative analysis of plate colony formation of 143B cells stably expressing miR-144. Silencing of miR-144 in Saos-2 cells significantly promoted proliferation E. and cell cycle G1/S transition F. *P < 0.05, **P < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4496356&req=5

Figure 2: miR-144 suppresses OS cell proliferation in vitroA. 143B cells were transiently transfected with miR-144 mimics or NC mimics, and cell viability determined with the CCK-8 assay. B. FACS analysis of 143B cells was performed after transfection with miR-144 mimics or NC mimics. C. The apoptosis cells were stained with Annexin V and PI, and analyzed using FACS. D. Representative photographs and quantitative analysis of plate colony formation of 143B cells stably expressing miR-144. Silencing of miR-144 in Saos-2 cells significantly promoted proliferation E. and cell cycle G1/S transition F. *P < 0.05, **P < 0.01.
Mentions: To explore the potential role of miR-144 in OS pathogenesis, 143B cells with high metastatic potential and low endogenous miR-144 expression, were transfected with miR-144 mimics, and miR-144 expression assayed using qRT-PCR (Supplementary Fig. 1A). Data from the cell viability assay showed that upregulation of miR-144 significantly suppresses the proliferation of 143B cells, compared with miR-NC-transfected cells (Fig. 2A). Flow cytometry analysis revealed that miR-144 overexpression leads to an increased percentage of cells in the G1 phase, alone with a decrease in S-phase cells (Fig. 2B), suggesting that this miRNA induces G1/S arrest. Moreover, the rate of apoptosis was significantly higher in 143B cells overexpressing miR-144 (Fig. 2C). The lentivirus system expressing miR-144-GFP was additionally applied to generate 143B-miR-144 cells stably expressing miR-144 (Supplementary Fig. 1B). As expected, the colony formation rate of these cells was significantly decreased (Fig. 2D).

Bottom Line: Low-level expression of miR-144 was significantly associated with distant metastasis and poor prognosis.Furthermore, we identified Rho-associated kinases 1 and 2 (ROCK1 and ROCK2) as direct targets for miR-144 binding, resulting in suppression of their expression.In clinical OS specimens, ROCK1 and ROCK2 levels were elevated, relative to that in paired normal bone tissues, and inversely correlated with miR-144 expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedic Surgery, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200127, China.

ABSTRACT
Osteosarcoma (OS) is the most common primary tumor of bone. MicroRNAs (miRNAs) are a class of endogenously expressed small non-coding RNAs that are strongly implicated in cancerous processes. However, our current understanding of the biological role of miRNAs in OS remains incomplete. In the present study, miR-144 was markedly downregulated in OS cell lines and clinical specimens. Low-level expression of miR-144 was significantly associated with distant metastasis and poor prognosis. Functional studies demonstrated that ectopic expression of miR-144 suppresses tumor cell proliferation and metastasis in vitro as well as in vivo. Furthermore, we identified Rho-associated kinases 1 and 2 (ROCK1 and ROCK2) as direct targets for miR-144 binding, resulting in suppression of their expression. Exogenous expression of ROCK1 or ROCK2 in 143B-miR-144 cells partially restored miR-144-inhibited cell proliferation and invasion. In clinical OS specimens, ROCK1 and ROCK2 levels were elevated, relative to that in paired normal bone tissues, and inversely correlated with miR-144 expression. Taken together, miR-144 suppresses OS progression by directly downregulating ROCK1 and ROCK2 expression, and may be a promising therapeutic target for OS.

No MeSH data available.


Related in: MedlinePlus