STANDARD_NAME BHATI_G2M_ARREST_BY_2METHOXYESTRADIOL_UP SYSTEMATIC_NAME M18550 COLLECTION C2:CGP MSIGDB_URL https://www.gsea-msigdb.org/gsea/msigdb/human/geneset/BHATI_G2M_ARREST_BY_2METHOXYESTRADIOL_UP NAMESPACE HUMAN_SEQ_ACCESSION DESCRIPTION_BRIEF Up-regulated genes in MDA-MB-435 cells (breast cancer) undergoing G2/M arrest after treatment with 2-methoxyestradiol (2ME2)[PubChem=1573]. DESCRIPTION_FULL 2-methoxyestradiol (2ME2), an estradiol metabolite with antiproliferative and antiangiogenic activities, is in phase I/II clinical trials for breast cancer. 2ME2 inhibits microtubule polymerization and causes cells to arrest in G2-M. The purpose of this study was to further elucidate the molecular mechanism of 2ME2. MDA-MB-435 breast cancer cells were treated with 2ME2 (2 micromol/L) or vehicle alone. RNA was extracted and genomic profiling was done using 22k Agilent microarrays. Expression Analysis Systematic Explorer was used to determine enrichment of Gene Ontology categories. Protein isolates were subjected to Western blot analysis. Protein synthesis was measured with a [35S]methionine pulse assay. An MDA-MB-435 cell line with two beta-tubulin mutations (2ME2R) was used to determine whether novel mechanisms were tubulin-dependent. Gene Ontology categories enriched include genes that regulate the mitotic spindle assembly checkpoint, apoptosis, and the cytosolic ribosome. The target of the mitotic spindle assembly checkpoint is the anaphase-promoting complex (APC). APC inhibition was confirmed by measuring protein levels of its targets securin and cyclin B1, which were increased in 2ME2-treated cells. Because gene expression in the cytosolic ribosome category was decreased, we evaluated whether 2ME2 decreases protein translation. This was confirmed with a pulse assay, which showed decreased isotope incorporation in 2ME2-treated cells, which was maintained in the tubulin-resistant 2ME2R cells. APC inhibition was not maintained in 2ME2R cells. 2ME2 induces tubulin-dependent cell cycle arrest through regulation of genes involved in the mitotic spindle assembly checkpoint, which results in inhibition of the APC and tubulin-independent inhibition of protein translation. PMID 17234781 GEOID GSE5665 AUTHORS Bhati R,Gökmen-Polar Y,Sledge GW Jr,Fan C,Nakshatri H,Ketelsen D,Borchers CH,Dial MJ,Patterson C,Klauber-DeMore N CONTRIBUTOR Jessica Robertson CONTRIBUTOR_ORG MSigDB Team EXACT_SOURCE Suppl. Gene List: Red FILTERED_BY_SIMILARITY EXTERNAL_NAMES_FOR_SIMILAR_TERMS EXTERNAL_DETAILS_URL SOURCE_MEMBERS AF086130,AF175767,AF229253,AK023831,AK024176,AK098134,AK125672,AK130644,AL359570,AL390173,AL832180,BC004399,BC006267,BC040697,BC044941,BC065722,NM_000636,NM_000700,NM_000923,NM_000937,NM_001254,NM_001255,NM_001502,NM_001554,NM_001559,NM_001905,NM_001964,NM_002061,NM_002110,NM_002270,NM_002314,NM_002333,NM_002353,NM_002358,NM_002466,NM_002497,NM_002524,NM_002592,NM_002687,NM_002787,NM_002788,NM_003090,NM_003118,NM_003198,NM_003234,NM_003534,NM_003600,NM_003798,NM_003807,NM_003897,NM_003937,NM_004094,NM_004126,NM_004268,NM_004336,NM_004343,NM_004460,NM_004481,NM_004482,NM_004580,NM_005002,NM_005003,NM_005252,NM_005319,NM_005347,NM_005348,NM_005389,NM_005855,NM_006607,NM_006850,NM_007045,NM_007080,NM_007110,NM_012242,NM_014325,NM_014504,NM_014750,NM_014888,NM_015277,NM_015459,NM_015472,NM_015895,NM_016205,NM_016297,NM_016395,NM_016587,NM_017805,NM_017852,NM_017934,NM_017958,NM_018029,NM_018087,NM_018136,NM_018466,NM_018492,NM_018991,NM_020390,NM_020675,NM_021052,NM_021190,NM_021928,NM_021972,NM_022766,NM_024576,NM_024895,NM_025049,NM_025143,NM_031427,NM_031966,NM_032105,NM_032391,NM_032525,NM_032753,NM_032895,NM_033066,NM_033224,NM_033557,NM_138957,NM_144653,NM_145697,NM_153338,NM_170743,NM_175748,NM_175839,NM_175924,NM_178510,U88965 GENE_SYMBOLS CALHM6,SCLY,API5,,SND1-IT1,,TMC8,,ANKRD42-DT,,ZSCAN30,TTC39A,ASRGL1,TBX18,FAM215B,BMS1P1,SOD2,ANXA1,PDE4C,POLR2A,CDC6,CDC20,GP2,CCN1,IL12RB2,CTPS1,EGR1,GCLM,HCK,TNPO1,LIMK1,LRP3,TACSTD2,MAD2L1,MYBL2,NEK2,NRAS,PCNA,PNN,PSMA2,PSMA3,SNRPA1,SPARC,ELOA,TFRC,H3C8,AURKA,CTNNAL1,TNFSF14,IER3,KYNU,EIF2S1,GNG11,MED17,BUB1,CALR,FAP,GALNT2,GALNT3,RAB27A,NDUFA9,NDUFAB1,FOS,H1-2,HSPA5,HSP90AA1,PCMT1,RAMP1,PTTG2,IL24,CEP43,LSM6,TEP1,DKK1,CORO1C,RABGEF1,DLGAP5,FAM3C,NEDD4L,ATL3,WWTR1,GMNN,PDGFC,PCYOX1,HACD3,CBX3,RASIP1,NLRP2,PHIP,PLEKHB2,EBLN2,NDC1,ASPM,ALG13,PBK,STAG3L1,EIF5A2,SPC25,H2AC8,PTBP2,SPCS3,SPHK1,CERK,OGFRL1,PDZD7,PIF1,,DNAL1,CCNB1,PPP1R12B,PRAC1,TUBB6,RAX2,MIR22HG,MPP4,PURB,YIF1B,MAPK1,NACC2,NUF2,GGT6,IFNLR1,UBR7,SMOX,ILDR1,ANKK1,ERC2-IT1 FOUNDER_NAMES