miR-221/222 Target the DNA Methyltransferase MGMT in Glioma Cells
Publication Date
September 19, 2013
Journal
PLOS ONE
Authors
Cristina Quintavalle, Davide Mangani, Giuseppina Roscigno, Giulia Romano, et al
Volume
8
Issue
9
Pages
e74466
DOI
https://dx.plos.org/10.1371/journal.pone.0074466
Publisher URL
http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0074466
PubMed
http://www.ncbi.nlm.nih.gov/pubmed/24147153
PubMed Central
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3798259
Europe PMC
http://europepmc.org/abstract/MED/24147153
Web of Science
000324777300035
Scopus
84884382332
Mendeley
http://www.mendeley.com/research/mir221222-target-dna-methyltransferase-mgmt-glioma-cells
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Mendeley | Further Information

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CrossRef

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1209744"], "description"=>"<div><p>Glioblastoma multiforme (GBM) is one of the most deadly types of cancer. To date, the best clinical approach for treatment is based on administration of temozolomide (TMZ) in combination with radiotherapy. Much evidence suggests that the intracellular level of the alkylating enzyme O<sup>6</sup>-methylguanine–DNA methyltransferase (MGMT) impacts response to TMZ in GBM patients. MGMT expression is regulated by the methylation of its promoter. However, evidence indicates that this is not the only regulatory mechanism present. Here, we describe a hitherto unknown microRNA-mediated mechanism of MGMT expression regulation. We show that miR-221 and miR-222 are upregulated in GMB patients and that these paralogues target MGMT mRNA, inducing greater TMZ-mediated cell death. However, miR-221/miR-222 also increase DNA damage and, thus, chromosomal rearrangements. Indeed, miR-221 overexpression in glioma cells led to an increase in markers of DNA damage, an effect rescued by re-expression of MGMT. Thus, chronic miR-221/222-mediated MGMT downregulation may render cells unable to repair genetic damage. This, associated also to miR-221/222 oncogenic potential, may poor GBM prognosis.</p> </div>", "links"=>[], "tags"=>["dna", "methyltransferase", "mgmt", "glioma"], "article_id"=>803509, "categories"=>["Biological Sciences"], "users"=>["Cristina Quintavalle", "Davide Mangani", "Giuseppina Roscigno", "Giulia Romano", "Angel Diaz-Lagares", "Margherita Iaboni", "Elvira Donnarumma", "Danilo Fiore", "Pasqualino De Marinis", "Ylermi Soini", "Manel Esteller", "Gerolama Condorelli"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0074466", "stats"=>{"downloads"=>1, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_miR_221_222_Target_the_DNA_Methyltransferase_MGMT_in_Glioma_Cells_/803509", "title"=>"miR-221/222 Target the DNA Methyltransferase MGMT in Glioma Cells", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-09-19 02:22:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1209743"], "description"=>"<p>Mann–Whitney U test analysis was performed to evaluate the association between miR-221 and MGMT expression in long- and short -survival groups of patients. The expression of miR-221 (2^-Dct) (<b>A</b>-<b>B</b>) and MGMT (2^-Dct) are inversely correlated with patient survival (<i>p</i> < 0.0490 and <i>p</i> = 0.043, respectively).</p>", "links"=>[], "tags"=>["mir-221", "mgmt"], "article_id"=>803508, "categories"=>["Biological Sciences"], "users"=>["Cristina Quintavalle", "Davide Mangani", "Giuseppina Roscigno", "Giulia Romano", "Angel Diaz-Lagares", "Margherita Iaboni", "Elvira Donnarumma", "Danilo Fiore", "Pasqualino De Marinis", "Ylermi Soini", "Manel Esteller", "Gerolama Condorelli"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0074466.g006", "stats"=>{"downloads"=>0, "page_views"=>22, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Association_of_miR_221_and_MGMT_expression_/803508", "title"=>"Association of miR-221 and MGMT expression.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-09-19 02:22:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1209738"], "description"=>"<p>(A) Western blot analysis and real time PCR of MGMT protein and RNA after miR-221/222 transfection of T98G cells. (B) Western blot analysis and real time PCR of MGMT protein and RNA after anti-miR-221 and -222 transfection of U87MG cells. (C) Western blot of MGMT expression upon miR-221 transfection of LN428 cells. (D) Western blot analysis of MGMT expression in T98G cells, as a control, and the melanoma cell line A375 upon miR-221 transfection. (E) Analysis of methylation status of MGMT promoter in T98G and U87MG upon miR- or anti-miR-221/222 transfection. U is for the un-methylated form, M for methylated form, NL is for normal lymphocytes, used as control.</p>", "links"=>[], "tags"=>[], "article_id"=>803503, "categories"=>["Biological Sciences"], "users"=>["Cristina Quintavalle", "Davide Mangani", "Giuseppina Roscigno", "Giulia Romano", "Angel Diaz-Lagares", "Margherita Iaboni", "Elvira Donnarumma", "Danilo Fiore", "Pasqualino De Marinis", "Ylermi Soini", "Manel Esteller", "Gerolama Condorelli"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0074466.g002", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_miR_221_222_target_MGMT_/803503", "title"=>"miR-221/222 target <i>MGMT</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-09-19 02:22:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1209739"], "description"=>"<p>(<b>A</b>) Cell viability of T98G, LN428, and A375 cells transfected with miR-221 and miR-222 upon TMZ treatment (300 µMol) for 24 hrs. **p value<0.0082 versus scr column, ***p value<0.005 versus scr column. (<b>B</b>) Growth curve of T98G and LN428 cells transfected or not with miR-221 after 24 hrs of treatment with TMZ. (<b>C</b>) Colony assay of T98G and LN428 cells transfected with miR-221 and then treated for 24 hrs with TMZ (300 µMol). Cells were left to grow for 6 days after treatment removal. (<b>D</b>) MGMT expression rescues cell viability after TMZ treatment in T98G and LN428 cells overexpressing miR-221 **p value<0.0082 versus untransfected MGMT column. (<b>E</b>) Correlation between miR-221 expression and TMZ sensitivity in nine primary glioblastoma cell lines and in six glioblastoma cell lines.</p>", "links"=>[], "tags"=>["modulates", "tmz"], "article_id"=>803504, "categories"=>["Biological Sciences"], "users"=>["Cristina Quintavalle", "Davide Mangani", "Giuseppina Roscigno", "Giulia Romano", "Angel Diaz-Lagares", "Margherita Iaboni", "Elvira Donnarumma", "Danilo Fiore", "Pasqualino De Marinis", "Ylermi Soini", "Manel Esteller", "Gerolama Condorelli"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0074466.g003", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_miR_221_modulates_TMZ_sensitivity_/803504", "title"=>"miR-221 modulates TMZ sensitivity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-09-19 02:22:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1209737"], "description"=>"<p>(A) Glioma cells were treated with TMZ (300µMol) for 24 hr. Cell viability was evaluated with an MTT assay. (B) Western blot analysis of MGMT expression in glioblastoma cells. (C) Real time PCR of miR-221 expression in glioblastoma cells. (D) RNA Hybrid prediction analyzes of miR-222, miR-221, and MGMT 3’ UTR. In bold are shown the mutated oligonucleotides. Luciferase activity of HEK-293 cells transiently co-transfected with the luciferase reporter containing wild-type MGMT-3’UTR or mutant MGMT-3’UTR in the presence of pre-miR-222, miR-221, or scrambled oligonucleotide. Representative of at least three independent experiments. *** p<0.001 versus control, ** p<0,0037 versus control.</p>", "links"=>[], "tags"=>["mgmt", "glioma"], "article_id"=>803502, "categories"=>["Biological Sciences"], "users"=>["Cristina Quintavalle", "Davide Mangani", "Giuseppina Roscigno", "Giulia Romano", "Angel Diaz-Lagares", "Margherita Iaboni", "Elvira Donnarumma", "Danilo Fiore", "Pasqualino De Marinis", "Ylermi Soini", "Manel Esteller", "Gerolama Condorelli"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0074466.g001", "stats"=>{"downloads"=>0, "page_views"=>25, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_TMZ_sensitivity_and_MGMT_and_miR_221_222_expression_in_glioma_cells_/803502", "title"=>"TMZ sensitivity and MGMT and miR-221/222 expression in glioma cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-09-19 02:22:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1209742"], "description"=>"<p>(A) Alkaline comet assay of T98G cells transfected with miR-221 and treated with TMZ for the indicated times. (B) Analysis of γH2AX in T98G cells transfected with scrambled control miR or miR-221, treated with TMZ in the presence or in the absence of MGMT cDNA, by immunocytofluorescence (upper and medium panel) or by Western blot (lower panel). (C) Western blot analysis of the indicated proteins upon transfection of T98G cells with miR-221 and MGMT cDNA and TMZ treatment for 24 hrs.</p>", "links"=>[], "tags"=>["dna"], "article_id"=>803507, "categories"=>["Biological Sciences"], "users"=>["Cristina Quintavalle", "Davide Mangani", "Giuseppina Roscigno", "Giulia Romano", "Angel Diaz-Lagares", "Margherita Iaboni", "Elvira Donnarumma", "Danilo Fiore", "Pasqualino De Marinis", "Ylermi Soini", "Manel Esteller", "Gerolama Condorelli"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0074466.g005", "stats"=>{"downloads"=>6, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_miR_221_promotes_DNA_damage_/803507", "title"=>"miR-221 promotes DNA damage.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-09-19 02:22:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1209741"], "description"=>"<p>(A) Apoptotic cell death assessed by FACS in T98G cells transfected with miR-221 or scrambled sequence and MGMT and treated with TMZ for 24 hrs. *** p value< 0.005 versus untrasfected MGMT column. (B) Active caspase-3 quantification in T98G cells as indicated and treated with TMZ for 24 hrs in the presence or absence of 3 hrs pre-treatment with ZVAD-fmk. (C) Upper panel Time course analysis of caspase-3 activation upon TMZ treatment in T98G cells transfected with miR-221 or with scrambled sequence. Lower panel Western blot analysis of caspase-3 activation after miR-221 and MGMT transfection. (D) Cell viability of T98G cells transfected with miR-221 or with scrambled sequence treated with TMZ for 24 hrs in the presence or absence of 3 hrs pre-treatment with ZVAD-fmk. ** p value< 0.0034 versus only treated TMZ column, Student’s t test.</p>", "links"=>[], "tags"=>["dna", "damages", "tmz"], "article_id"=>803506, "categories"=>["Biological Sciences"], "users"=>["Cristina Quintavalle", "Davide Mangani", "Giuseppina Roscigno", "Giulia Romano", "Angel Diaz-Lagares", "Margherita Iaboni", "Elvira Donnarumma", "Danilo Fiore", "Pasqualino De Marinis", "Ylermi Soini", "Manel Esteller", "Gerolama Condorelli"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0074466.g004", "stats"=>{"downloads"=>5, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_miR_221_promotes_DNA_damages_upon_TMZ_treatment_/803506", "title"=>"miR-221 promotes DNA damages upon TMZ treatment.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-09-19 02:22:49"}

PMC Usage Stats | Further Information

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  • {"unique-ip"=>"5", "full-text"=>"5", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"8"}
  • {"unique-ip"=>"12", "full-text"=>"20", "pdf"=>"6", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2018", "month"=>"10"}
  • {"unique-ip"=>"6", "full-text"=>"6", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"11"}
  • {"unique-ip"=>"10", "full-text"=>"12", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"9"}
  • {"unique-ip"=>"4", "full-text"=>"4", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"2"}
  • {"unique-ip"=>"8", "full-text"=>"11", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"7", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"3"}
  • {"unique-ip"=>"11", "full-text"=>"21", "pdf"=>"9", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"2", "cited-by"=>"0", "year"=>"2019", "month"=>"4"}
  • {"unique-ip"=>"11", "full-text"=>"10", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"5"}
  • {"unique-ip"=>"12", "full-text"=>"11", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"8"}
  • {"unique-ip"=>"6", "full-text"=>"5", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"1", "year"=>"2019", "month"=>"9"}
  • {"unique-ip"=>"16", "full-text"=>"14", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"5", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"10"}

Relative Metric

{"start_date"=>"2013-01-01T00:00:00Z", "end_date"=>"2013-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Biology and life sciences", "average_usage"=>[269, 466, 588, 697, 800, 896, 988, 1076, 1165, 1254, 1340, 1417]}, {"subject_area"=>"/Biology and life sciences/Biochemistry", "average_usage"=>[266, 468, 593, 703, 804, 903, 993, 1084, 1171, 1256, 1339, 1422, 1492]}, {"subject_area"=>"/Biology and life sciences/Cell biology", "average_usage"=>[272, 472, 600, 713, 815, 911, 1004, 1094, 1185, 1273, 1358, 1441]}, {"subject_area"=>"/Biology and life sciences/Genetics", "average_usage"=>[284, 491, 620, 738, 843, 945, 1043, 1137, 1225, 1315, 1400, 1479, 1555]}, {"subject_area"=>"/Biology and life sciences/Molecular biology", "average_usage"=>[272, 466, 589, 702, 806, 903, 995, 1086, 1176, 1258, 1347, 1422, 1493]}]}
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