MicroRNA-145 Regulates Chondrogenic Differentiation of Mesenchymal Stem Cells by Targeting Sox9
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{"title"=>"Microrna-145 regulates chondrogenic differentiation of mesenchymal stem cells by targeting SOX9", "type"=>"journal", "authors"=>[{"first_name"=>"Bo", "last_name"=>"Yang", "scopus_author_id"=>"7404472939"}, {"first_name"=>"Hongfeng", "last_name"=>"Guo", "scopus_author_id"=>"55468669500"}, {"first_name"=>"Yulan", "last_name"=>"Zhang", "scopus_author_id"=>"38663790900"}, {"first_name"=>"Lei", "last_name"=>"Chen", "scopus_author_id"=>"57190219800"}, {"first_name"=>"Dajun", "last_name"=>"Ying", "scopus_author_id"=>"7005427206"}, {"first_name"=>"Shiwu", "last_name"=>"Dong", "scopus_author_id"=>"24757922900"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"79960569238", "doi"=>"10.1371/journal.pone.0021679", "pui"=>"362184563", "pmid"=>"21799743", "scopus"=>"2-s2.0-79960569238", "issn"=>"19326203", "isbn"=>"1932-6203"}, "id"=>"20e0f882-71fe-39fa-b69f-ad2cc6f46538", "abstract"=>"Chondrogenic differentiation of mesenchymal stem cells (MSCs) is accurately regulated by essential transcription factors and signaling cascades. However, the precise mechanisms involved in this process still remain to be defined. MicroRNAs (miRNAs) regulate various biological processes by binding target mRNA to attenuate protein synthesis. To investigate the mechanisms for miRNAs-mediated regulation of chondrogenic differentiation, we identified that miR-145 was decreased during transforming growth factor beta 3 (TGF-β3)-induced chondrogenic differentiation of murine MSCs. Subsequently, dual-luciferase reporter gene assay data demonstrated that miR-145 targets a putative binding site in the 3'-UTR of SRY-related high mobility group-Box gene 9 (Sox9) gene, the key transcription factor for chondrogenesis. In addition, over-expression of miR-145 decreased expression of Sox9 only at protein levels and miR-145 inhibition significantly elevated Sox9 protein levels. Furthermore, over-expression of miR-145 decreased mRNA levels for three chondrogenic marker genes, type II collagen (Col2a1), aggrecan (Agc1), cartilage oligomeric matrix protein (COMP), type IX collagen (Col9a2) and type XI collagen (Col11a1) in C3H10T1/2 cells induced by TGF-β3, whereas anti-miR-145 inhibitor increased the expression of these chondrogenic marker genes. Thus, our studies demonstrated that miR-145 is a key negative regulator of chondrogenic differentiation by directly targeting Sox9 at early stage of chondrogenic differentiation.", "link"=>"http://www.mendeley.com/research/microrna145-regulates-chondrogenic-differentiation-mesenchymal-stem-cells-targeting-sox9-3", "reader_count"=>53, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Professor > Associate Professor"=>2, "Researcher"=>8, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>22, "Student > Master"=>12, "Other"=>1, "Student > Bachelor"=>2, "Lecturer"=>1, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Professor > Associate Professor"=>2, "Researcher"=>8, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>22, "Student > Master"=>12, "Other"=>1, "Student > Bachelor"=>2, "Lecturer"=>1, "Professor"=>2}, "reader_count_by_subject_area"=>{"Engineering"=>5, "Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>7, "Medicine and Dentistry"=>10, "Agricultural and Biological Sciences"=>26, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Chemistry"=>1, "Economics, Econometrics and Finance"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>5}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>10}, "Chemistry"=>{"Chemistry"=>1}, "Economics, Econometrics and Finance"=>{"Economics, Econometrics and Finance"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>26}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>7}, "Unspecified"=>{"Unspecified"=>2}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"United States"=>1, "Japan"=>1, "Chile"=>1, "India"=>1}, "group_count"=>2}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/379748"], "description"=>"<div><p>Chondrogenic differentiation of mesenchymal stem cells (MSCs) is accurately regulated by essential transcription factors and signaling cascades. However, the precise mechanisms involved in this process still remain to be defined. MicroRNAs (miRNAs) regulate various biological processes by binding target mRNA to attenuate protein synthesis. To investigate the mechanisms for miRNAs-mediated regulation of chondrogenic differentiation, we identified that miR-145 was decreased during transforming growth factor beta 3 (TGF-β3)-induced chondrogenic differentiation of murine MSCs. Subsequently, dual-luciferase reporter gene assay data demonstrated that miR-145 targets a putative binding site in the 3′-UTR of SRY-related high mobility group-Box gene 9 (Sox9) gene, the key transcription factor for chondrogenesis. In addition, over-expression of miR-145 decreased expression of Sox9 only at protein levels and miR-145 inhibition significantly elevated Sox9 protein levels. Furthermore, over-expression of miR-145 decreased mRNA levels for three chondrogenic marker genes, type II collagen (Col2a1), aggrecan (Agc1), cartilage oligomeric matrix protein (COMP), type IX collagen (Col9a2) and type XI collagen (Col11a1) in C3H10T1/2 cells induced by TGF-β3, whereas anti-miR-145 inhibitor increased the expression of these chondrogenic marker genes. Thus, our studies demonstrated that miR-145 is a key negative regulator of chondrogenic differentiation by directly targeting Sox9 at early stage of chondrogenic differentiation.</p> </div>", "links"=>[], "tags"=>["microrna-145", "regulates", "chondrogenic", "differentiation", "mesenchymal", "cells", "targeting", "sox9"], "article_id"=>135040, "categories"=>["Molecular Biology", "Biochemistry", "Cell Biology", "Genetics"], "users"=>["Bo Yang", "Hongfeng Guo", "Yulan Zhang", "Lei Chen", "Dajun Ying", "Shiwu Dong"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021679", "stats"=>{"downloads"=>1, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/MicroRNA_145_Regulates_Chondrogenic_Differentiation_of_Mesenchymal_Stem_Cells_by_Targeting_Sox9/135040", "title"=>"MicroRNA-145 Regulates Chondrogenic Differentiation of Mesenchymal Stem Cells by Targeting Sox9", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-20 01:24:00"}
  • {"files"=>["https://ndownloader.figshare.com/files/754571"], "description"=>"<p>(A) A schematic shows that the 5′ end of miR-145 contains a sequence complementary to the specific miRNA binding site within the 3′-UTR of Sox9 mRNA. (B) Pre-miR-145 or its negative control was co-transfected with the specific pMIR-REPORT construct containing a consensus miR-145-binding site (pMIR-PT) into HEK293 cells. Pre-miR-145 or its negative control was co-transfected with pMIR-MRE into HEK293 cells. pMIR-PT acts as a positive control. (C) HEK293 cells were co-transfected with pMIR-MRE together with varying amounts of pre-miR-145, as indicated. Statistical comparisons were made between multiple groups by ANOVA. (D) Anti-miR-145 or its negative control was co-transfected with either pre-miR-145 together with pMIR-MRE or pre-miR-145 together with pMIR-MUT into HEK293 cells. pMIR-PT acts as a positive control. All cells (B, C, D) were harvested at 48 h after transfection, and then luciferase activities were measured and normalized to the phRL-TK activities. Three independent transfection experiments were done and data was represented as mean±sd. *, <i>p</i><0.05, **, <i>p</i><0.01, when compared with control.</p>", "links"=>[], "tags"=>["targets"], "article_id"=>424941, "categories"=>["Molecular Biology", "Biochemistry", "Cell Biology", "Genetics"], "users"=>["Bo Yang", "Hongfeng Guo", "Yulan Zhang", "Lei Chen", "Dajun Ying", "Shiwu Dong"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021679.g002", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mir_145_directly_targets_Sox9_/424941", "title"=>"Mir-145 directly targets Sox9.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-20 01:22:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/754810"], "description"=>"<p>C3H10T1/2 cells were transfected with pre-miR-145 or its control respectively. (A) After 24 h, 7 d and 14 d of treatment with TGF-β3, all of cells were lysed and then the expression of chondrogenic differentiation markers, such as Col2a1, COMP, Agc1, Col9a2 and Col11a1, were measured via qRT-PCR. The relative expression level of mRNA in cells transfected with control oligonucleotide was set to one, as control. (B) After 3 d, 7 d and 14 d of treatment with TGF-β3, all of pellets were measured by alcian blue staining. Three independent experiments were done and data was represented as mean±sd. *, <i>p</i><0.05, when compared with control.</p>", "links"=>[], "tags"=>["mir-145", "inhibits", "chondrogenic", "differentiation"], "article_id"=>425178, "categories"=>["Molecular Biology", "Biochemistry", "Cell Biology", "Genetics"], "users"=>["Bo Yang", "Hongfeng Guo", "Yulan Zhang", "Lei Chen", "Dajun Ying", "Shiwu Dong"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021679.g004", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Over_expression_of_mir_145_inhibits_early_chondrogenic_differentiation_of_C3H10T1_2_cells_/425178", "title"=>"Over-expression of mir-145 inhibits early chondrogenic differentiation of C3H10T1/2 cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-20 01:26:18"}
  • {"files"=>["https://ndownloader.figshare.com/files/754951"], "description"=>"<p>C3H10T1/2 cells were transfected with anti-miR-145 or its control respectively. (A) After 24 h, 7 d and 14 d of treatment with TGF-β3, all of cells were lysed and then the expression of chondrogenic differentiation markers, such as Col2a1, COMP, Agc1, Col9a2 and Col11a1, were measured via qRT-PCR. The relative expression level of mRNA in cells transfected with control oligonucleotide was set to one, as control. (B) After 3 d, 7 d and 14d of treatment with TGF-β3, all of pellets were measured by alcian blue staining. Three independent cell culture experiments were done and data was represented as mean±sd. *, <i>p</i><0.05, when compared with control.</p>", "links"=>[], "tags"=>["mir-145", "enhances", "chondrogenic", "differentiation"], "article_id"=>425319, "categories"=>["Molecular Biology", "Biochemistry", "Cell Biology", "Genetics"], "users"=>["Bo Yang", "Hongfeng Guo", "Yulan Zhang", "Lei Chen", "Dajun Ying", "Shiwu Dong"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021679.g005", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Suppression_of_mir_145_enhances_early_chondrogenic_differentiation_of_C3H10T1_2_cells_/425319", "title"=>"Suppression of mir-145 enhances early chondrogenic differentiation of C3H10T1/2 cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-20 01:28:39"}
  • {"files"=>["https://ndownloader.figshare.com/files/754681"], "description"=>"<p>(A) C3H10T1/2 cells were transfected with pre-miR-145, anti-miR-145 or their control individually. After induced by TGF-β3 for 24 h, 7 d and 14 d, the cells were harvested for measurement of Sox9 protein expression using Western blot. β-actin acts as an internal control. Quantitation of the Sox9 protein level was performed using Quantity One software. The result is shown in the below panels. (B) C3H10T1/2 cells were transfected with pre-miR-145, anti-miR-145 or their control, and then mRNA level of Sox9 were measured using qRT-PCR at 24h. β-actin acts as an internal control in qRT-PCR analysis. The relative expression level of Sox9 mRNA in cells transfected with control oligonucleotide was set to one, as control. Three independent experiments were done and data was represented as mean±sd.</p>", "links"=>[], "tags"=>["represses", "sox9", "chondrogenic"], "article_id"=>425053, "categories"=>["Molecular Biology", "Biochemistry", "Cell Biology", "Genetics"], "users"=>["Bo Yang", "Hongfeng Guo", "Yulan Zhang", "Lei Chen", "Dajun Ying", "Shiwu Dong"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021679.g003", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mir_145_represses_the_expression_of_Sox9_at_protein_level_in_early_stage_of_chondrogenic_differentiation_/425053", "title"=>"Mir-145 represses the expression of Sox9 at protein level in early stage of chondrogenic differentiation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-20 01:24:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/755160"], "description"=>"<p>C3H10T1/2 cells were transfected with pre-miR145, anti-miR-145 or its control, respectively. After 24 h of treatment with TGF-β3, all of pellets were trypsinized and directly counted in triplicate using a hemacytometer. Data was represented as mean±sd.</p>", "links"=>[], "tags"=>["proliferation"], "article_id"=>425532, "categories"=>["Molecular Biology", "Biochemistry", "Cell Biology", "Genetics"], "users"=>["Bo Yang", "Hongfeng Guo", "Yulan Zhang", "Lei Chen", "Dajun Ying", "Shiwu Dong"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021679.g007", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mir_145_has_no_influence_on_proliferation_of_C3H10T1_2_cells_/425532", "title"=>"Mir-145 has no influence on proliferation of C3H10T1/2 cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-20 01:32:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/754512"], "description"=>"<p>Down-regulated expression of miR-145 in murine MSCs induced by TGF-β3 was identified by qRT-PCR at different stage of chondrogenic differentiation (0 d, 7 d, 14 d). The relative expression level of miR-145 in untreated MSCs (0 d) was set to one, as control. Three independent cell culture experiments were done and data was represented as mean±sd. *, <i>p</i><0.05, when compared with control.</p>", "links"=>[], "tags"=>["mir-145", "chondrogenic", "differentiation", "murine"], "article_id"=>424874, "categories"=>["Molecular Biology", "Biochemistry", "Cell Biology", "Genetics"], "users"=>["Bo Yang", "Hongfeng Guo", "Yulan Zhang", "Lei Chen", "Dajun Ying", "Shiwu Dong"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021679.g001", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Differential_expression_of_miR_145_during_chondrogenic_differentiation_of_murine_MSCs_/424874", "title"=>"Differential expression of miR-145 during chondrogenic differentiation of murine MSCs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-20 01:21:14"}
  • {"files"=>["https://ndownloader.figshare.com/files/755224"], "description"=>"<p>The sequences of linker are in lowercase.</p>", "links"=>[], "tags"=>["oligonucleotides", "sequences", "luciferase"], "article_id"=>425592, "categories"=>["Molecular Biology", "Biochemistry", "Cell Biology", "Genetics"], "users"=>["Bo Yang", "Hongfeng Guo", "Yulan Zhang", "Lei Chen", "Dajun Ying", "Shiwu Dong"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021679.t001", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Synthesized_oligonucleotides_sequences_for_generation_of_luciferase_reporter_constructs_/425592", "title"=>"Synthesized oligonucleotides sequences for generation of luciferase reporter constructs.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2011-07-20 01:33:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/755053"], "description"=>"<p>C3H10T1/2 cells were transfected with pre-miR145, anti-miR-145 or its control, respectively. After 24 h of treatment with TGF-β3, all of cells were lysed. The expression of C/EBPβ and C/EBPδ were measured via qRT-PCR. The relative expression level of mRNA in cells transfected with control oligonucleotide was set to one, as control. Three independent cell culture experiments were done and data was represented as mean±sd. *, <i>p</i><0.05, when compared with control.</p>", "links"=>[], "tags"=>["mrna"], "article_id"=>425427, "categories"=>["Molecular Biology", "Biochemistry", "Cell Biology", "Genetics"], "users"=>["Bo Yang", "Hongfeng Guo", "Yulan Zhang", "Lei Chen", "Dajun Ying", "Shiwu Dong"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021679.g006", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mir_145_has_no_influence_on_mRNA_expression_of_C_EBP_946_and_C_EBP_948_/425427", "title"=>"Mir-145 has no influence on mRNA expression of C/EBPβ and C/EBPδ.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-20 01:30:27"}

PMC Usage Stats | Further Information

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Relative Metric

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