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n2:pmcid: PMC0
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Subject Item: _:vb12282576
rdf:type: n5:Section
dc:title: summary and future directions
n5:contains: _:vb12282577 _:vb12282578

Subject Item: _:vb12282577
rdf:type: n2:Context
rdf:value: Chromosome instability is found in the majority of colon cancers, resulting primarily from deregulation of the DNA replication and mitotic spindle checkpoints (reviewed in [>>45<<]). Genes involved in canonical HH signaling have been linked to genomic instability, involving inactivation of DNA repair mechanisms, defects in checkpoint activation, and predisposition to development of cancers[46-49]. It therefore
n2:mentions: n3:15000146

Subject Item: _:vb12282578
rdf:type: n2:Context
rdf:value: Genes involved in canonical HH signaling have been linked to genomic instability, involving inactivation of DNA repair mechanisms, defects in checkpoint activation, and predisposition to development of cancers[>>46<<-49]. It therefore follows that termination of HH signaling at the level of GLI may constitute a critical event in determining the balance between cell survival and cell death. The combination of molecular and cellular approaches that are
n2:mentions: n3:19164512

Subject Item: _:vb12282579
rdf:type: n5:Section
dc:title: canonical hedgehog signaling in cancer
n5:contains: _:vb12282600 _:vb12282592 _:vb12282593 _:vb12282594 _:vb12282595 _:vb12282596 _:vb12282597 _:vb12282598 _:vb12282599 _:vb12282584 _:vb12282585 _:vb12282586 _:vb12282587 _:vb12282588 _:vb12282589 _:vb12282590 _:vb12282591 _:vb12282580 _:vb12282581 _:vb12282582 _:vb12282583

Subject Item: _:vb12282580
rdf:type: n2:Context
rdf:value: Canonical HH signaling engages PTCH, SMO and the GLI family of transcription factors (Figure 1), and in normal cellular processes is involved in embryogenesis, tissue patterning, stem cell function, and differentiation[>>1<<, 2]. Several types of human cancers have demonstrated aberrant activation of the HH pathway by ligand-independent signaling such as, amplification of GLI1 or GLI2, mutations in PTCH or SMO, or dysregulated gene expression[1, 3].
n2:mentions: n3:19860666

Subject Item: _:vb12282581
rdf:type: n2:Context
rdf:value: Canonical HH signaling engages PTCH, SMO and the GLI family of transcription factors (Figure 1), and in normal cellular processes is involved in embryogenesis, tissue patterning, stem cell function, and differentiation[1, >>2<<]. Several types of human cancers have demonstrated aberrant activation of the HH pathway by ligand-independent signaling such as, amplification of GLI1 or GLI2, mutations in PTCH or SMO, or dysregulated gene expression[1, 3].
n2:mentions: n3:19095649

Subject Item: _:vb12282582
rdf:type: n2:Context
rdf:value: Several types of human cancers have demonstrated aberrant activation of the HH pathway by ligand-independent signaling such as, amplification of GLI1 or GLI2, mutations in PTCH or SMO, or dysregulated gene expression[>>1<<, 3]. In colon cancer, aberrant HH signaling progresses during carcinogenesis and in metastatic disease[4-6], and is also activated in human colon carcinoma cell lines[7-9] and xenograft models[4], by ligand-dependent activation, that
n2:mentions: n3:19860666

Subject Item: _:vb12282583
rdf:type: n2:Context
rdf:value: Several types of human cancers have demonstrated aberrant activation of the HH pathway by ligand-independent signaling such as, amplification of GLI1 or GLI2, mutations in PTCH or SMO, or dysregulated gene expression[1, >>3<<]. In colon cancer, aberrant HH signaling progresses during carcinogenesis and in metastatic disease[4-6], and is also activated in human colon carcinoma cell lines[7-9] and xenograft models[4], by ligand-dependent activation, that occurs
n2:mentions: n3:17845852

Subject Item: _:vb12282584
rdf:type: n2:Context
rdf:value: In colon cancer, aberrant HH signaling progresses during carcinogenesis and in metastatic disease[>>4<<-6], and is also activated in human colon carcinoma cell lines[7-9] and xenograft models[4], by ligand-dependent activation, that occurs in GI cancers[1, 10].
n2:mentions: n3:19662327 n3:20049737 n3:17461467

Subject Item: _:vb12282585
rdf:type: n2:Context
rdf:value: In colon cancer, aberrant HH signaling progresses during carcinogenesis and in metastatic disease[4-6], and is also activated in human colon carcinoma cell lines[>>7<<-9] and xenograft models[4], by ligand-dependent activation, that occurs in GI cancers[1, 10].
n2:mentions: n3:20399136 n3:21135115

Subject Item: _:vb12282586
rdf:type: n2:Context
rdf:value: In colon cancer, aberrant HH signaling progresses during carcinogenesis and in metastatic disease[4-6], and is also activated in human colon carcinoma cell lines[7-9] and xenograft models[>>4<<], by ligand-dependent activation, that occurs in GI cancers[1, 10].
n2:mentions: n3:20049737

Subject Item: _:vb12282587
rdf:type: n2:Context
rdf:value: aberrant HH signaling progresses during carcinogenesis and in metastatic disease[4-6], and is also activated in human colon carcinoma cell lines[7-9] and xenograft models[4], by ligand-dependent activation, that occurs in GI cancers[>>1<<, 10]. However, the role of HH signaling and its importance in driving cellular survival in colon cancer are not well defined.
n2:mentions: n3:19860666

Subject Item: _:vb12282588
rdf:type: n2:Context
rdf:value: aberrant HH signaling progresses during carcinogenesis and in metastatic disease[4-6], and is also activated in human colon carcinoma cell lines[7-9] and xenograft models[4], by ligand-dependent activation, that occurs in GI cancers[1, >>10<<]. However, the role of HH signaling and its importance in driving cellular survival in colon cancer are not well defined.
n2:mentions: n3:21041712

Subject Item: _:vb12282589
rdf:type: n2:Context
rdf:value: Small molecule inhibitors of SMO have been studied in preclinical models, and applied to the treatment of various types of cancers in humans[>>4<<, 9, 11-14].
n2:mentions: n3:20049737

Subject Item: _:vb12282590
rdf:type: n2:Context
rdf:value: Small molecule inhibitors of SMO have been studied in preclinical models, and applied to the treatment of various types of cancers in humans[4, >>9<<, 11-14]. Those tumors sensitive to SMO inhibitors, which include basal cell carcinoma[15, 16] and medulloblastoma[11, 17], rely on canonical HH signaling for cellular survival. In other cancer types, SMO inhibitors including GDC-0449,
n2:mentions: n3:20399136

Subject Item: _:vb12282591
rdf:type: n2:Context
rdf:value: Small molecule inhibitors of SMO have been studied in preclinical models, and applied to the treatment of various types of cancers in humans[4, 9, >>11<<-14]. Those tumors sensitive to SMO inhibitors, which include basal cell carcinoma[15, 16] and medulloblastoma[11, 17], rely on canonical HH signaling for cellular survival. In other cancer types, SMO inhibitors including GDC-0449, IPI-926
n2:mentions: n3:21300762 n3:19726763 n3:20412786

Subject Item: _:vb12282592
rdf:type: n2:Context
rdf:value: Those tumors sensitive to SMO inhibitors, which include basal cell carcinoma[>>15<<, 16] and medulloblastoma[11, 17], rely on canonical HH signaling for cellular survival.
n2:mentions: n3:20072843

Subject Item: _:vb12282593
rdf:type: n2:Context
rdf:value: Those tumors sensitive to SMO inhibitors, which include basal cell carcinoma[15, >>16<<] and medulloblastoma[11, 17], rely on canonical HH signaling for cellular survival.
n2:mentions: n3:19726761

Subject Item: _:vb12282594
rdf:type: n2:Context
rdf:value: Those tumors sensitive to SMO inhibitors, which include basal cell carcinoma[15, 16] and medulloblastoma[>>11<<, 17], rely on canonical HH signaling for cellular survival.
n2:mentions: n3:20412786

Subject Item: _:vb12282595
rdf:type: n2:Context
rdf:value: Those tumors sensitive to SMO inhibitors, which include basal cell carcinoma[15, 16] and medulloblastoma[11, >>17<<], rely on canonical HH signaling for cellular survival.
n2:mentions: n3:17353198

Subject Item: _:vb12282596
rdf:type: n2:Context
rdf:value: In other cancer types, SMO inhibitors including GDC-0449, IPI-926 or LDE225, have demonstrated limited clinical activity (reviewed in [>>11<<, 12]). Intrinsic resistance to SMO inhibitors is frequent[11-14, 18, 19], and acquired resistance to GDC-0449 following initial response has been reported in medulloblastoma (heterozygous mutation, Asp->His at aa 473 in SMO)[20]. Thus
n2:mentions: n3:20412786

Subject Item: _:vb12282597
rdf:type: n2:Context
rdf:value: Intrinsic resistance to SMO inhibitors is frequent[>>11<<-14, 18, 19], and acquired resistance to GDC-0449 following initial response has been reported in medulloblastoma (heterozygous mutation, Asp->His at aa 473 in SMO)[20].
n2:mentions: n3:21300762 n3:19726763 n3:20412786

Subject Item: _:vb12282598
rdf:type: n2:Context
rdf:value: Intrinsic resistance to SMO inhibitors is frequent[11-14, >>18<<, 19], and acquired resistance to GDC-0449 following initial response has been reported in medulloblastoma (heterozygous mutation, Asp->His at aa 473 in SMO)[20].
n2:mentions: n3:17392427

Subject Item: _:vb12282599
rdf:type: n2:Context
rdf:value: Intrinsic resistance to SMO inhibitors is frequent[11-14, 18, >>19<<], and acquired resistance to GDC-0449 following initial response has been reported in medulloblastoma (heterozygous mutation, Asp->His at aa 473 in SMO)[20].
n2:mentions: n3:19726788

Subject Item: _:vb12282600
rdf:type: n2:Context
rdf:value: Intrinsic resistance to SMO inhibitors is frequent[11-14, 18, 19], and acquired resistance to GDC-0449 following initial response has been reported in medulloblastoma (heterozygous mutation, Asp->His at aa 473 in SMO)[>>20<<]. Thus targeting the GLI genes downstream of SMO, that constitute the core of HH-dependent gene regulation, may provide a significant advantage in eliminating HH signaling.
n2:mentions: n3:19190345

Subject Item: _:vb12282601
rdf:type: n5:Section
dc:title: activation of gli by oncogenic, non-canonical signaling pathways
n5:contains: _:vb12282602 _:vb12282603 _:vb12282604 _:vb12282605 _:vb12282606 _:vb12282607 _:vb12282608 _:vb12282609 _:vb12282610 _:vb12282611 _:vb12282612 _:vb12282613

Subject Item: _:vb12282602
rdf:type: n2:Context
rdf:value: Non-canonical, oncogene-driven signaling pathways converge on the activation of GLI genes and further converge on their specific downstream targets[>>3<<, 18, 21, 22] (see Figure 1). The RAS/RAF/MEK/ERK pathway, with activating mutations in K-RAS or B-RAF that occur in high frequency in colon cancers[23-25], activates GLI function[18, 19, 21].
n2:mentions: n3:17845852

Subject Item: _:vb12282603
rdf:type: n2:Context
rdf:value: Non-canonical, oncogene-driven signaling pathways converge on the activation of GLI genes and further converge on their specific downstream targets[3, >>18<<, 21, 22] (see Figure 1). The RAS/RAF/MEK/ERK pathway, with activating mutations in K-RAS or B-RAF that occur in high frequency in colon cancers[23-25], activates GLI function[18, 19, 21].
n2:mentions: n3:17392427

Subject Item: _:vb12282604
rdf:type: n2:Context
rdf:value: Non-canonical, oncogene-driven signaling pathways converge on the activation of GLI genes and further converge on their specific downstream targets[3, 18, >>21<<, 22] (see Figure 1). The RAS/RAF/MEK/ERK pathway, with activating mutations in K-RAS or B-RAF that occur in high frequency in colon cancers[23-25], activates GLI function[18, 19, 21].
n2:mentions: n3:11719506

Subject Item: _:vb12282605
rdf:type: n2:Context
rdf:value: Non-canonical, oncogene-driven signaling pathways converge on the activation of GLI genes and further converge on their specific downstream targets[3, 18, 21, >>22<<] (see Figure 1). The RAS/RAF/MEK/ERK pathway, with activating mutations in K-RAS or B-RAF that occur in high frequency in colon cancers[23-25], activates GLI function[18, 19, 21].
n2:mentions: n3:20071686

Subject Item: _:vb12282606
rdf:type: n2:Context
rdf:value: The RAS/RAF/MEK/ERK pathway, with activating mutations in K-RAS or B-RAF that occur in high frequency in colon cancers[>>23<<-25], activates GLI function[18, 19, 21].
n2:mentions: n3:12781369 n3:19258428 n3:20027224

Subject Item: _:vb12282607
rdf:type: n2:Context
rdf:value: The RAS/RAF/MEK/ERK pathway, with activating mutations in K-RAS or B-RAF that occur in high frequency in colon cancers[23-25], activates GLI function[>>18<<, 19, 21]. In HT29 cells (mutated B-RAF V600E[25]), we demonstrated inhibition of GLI-luciferase reporter activity, reduced expression of GLI1 mRNA and protein, and of p-ERK in response to the MEK/ERK and RAS/RAF signaling inhibitor
n2:mentions: n3:17392427

Subject Item: _:vb12282608
rdf:type: n2:Context
rdf:value: The RAS/RAF/MEK/ERK pathway, with activating mutations in K-RAS or B-RAF that occur in high frequency in colon cancers[23-25], activates GLI function[18, >>19<<, 21]. In HT29 cells (mutated B-RAF V600E[25]), we demonstrated inhibition of GLI-luciferase reporter activity, reduced expression of GLI1 mRNA and protein, and of p-ERK in response to the MEK/ERK and RAS/RAF signaling inhibitor U0126[26,
n2:mentions: n3:19726788

Subject Item: _:vb12282609
rdf:type: n2:Context
rdf:value: The RAS/RAF/MEK/ERK pathway, with activating mutations in K-RAS or B-RAF that occur in high frequency in colon cancers[23-25], activates GLI function[18, 19, >>21<<]. In HT29 cells (mutated B-RAF V600E[25]), we demonstrated inhibition of GLI-luciferase reporter activity, reduced expression of GLI1 mRNA and protein, and of p-ERK in response to the MEK/ERK and RAS/RAF signaling inhibitor U0126[26, 27]
n2:mentions: n3:11719506

Subject Item: _:vb12282610
rdf:type: n2:Context
rdf:value: In HT29 cells (mutated B-RAF V600E[>>25<<]), we demonstrated inhibition of GLI-luciferase reporter activity, reduced expression of GLI1 mRNA and protein, and of p-ERK in response to the MEK/ERK and RAS/RAF signaling inhibitor U0126[26, 27] (Figure 2).
n2:mentions: n3:19258428

Subject Item: _:vb12282611
rdf:type: n2:Context
rdf:value: In HT29 cells (mutated B-RAF V600E[25]), we demonstrated inhibition of GLI-luciferase reporter activity, reduced expression of GLI1 mRNA and protein, and of p-ERK in response to the MEK/ERK and RAS/RAF signaling inhibitor U0126[>>26<<, 27] (Figure 2).
n2:mentions: n3:17172409

Subject Item: _:vb12282612
rdf:type: n2:Context
rdf:value: In HT29 cells (mutated B-RAF V600E[25]), we demonstrated inhibition of GLI-luciferase reporter activity, reduced expression of GLI1 mRNA and protein, and of p-ERK in response to the MEK/ERK and RAS/RAF signaling inhibitor U0126[26, >>27<<] (Figure 2).
n2:mentions: n3:18006803

Subject Item: _:vb12282613
rdf:type: n2:Context
rdf:value: While loss-of-function mutations in PTCH and gain-of-function mutations in SMO activate HH signaling[>>1<<], acquired mutations in SMO or non-canonical GLI activation render cancer cells resistant to SMO antagonists.
n2:mentions: n3:19860666

Subject Item: _:vb12282614
rdf:type: n5:Section
dc:title: targeting gli1 and gli2 with gant61
n5:contains: _:vb12282620 _:vb12282621 _:vb12282616 _:vb12282617 _:vb12282618 _:vb12282619 _:vb12282615

Subject Item: _:vb12282615
rdf:type: n2:Context
rdf:value: GLI1 and GLI2 are the primary activators of HH signaling; further, the cooperative roles of GLI1 and GLI2 are critical in the transcriptional regulation of HH target genes[>>1<<, 28-30]. While SMO has been extensively investigated as a therapeutic target[11-15, 17], few agents are available that target the GLI genes[31]. GANT61 was identified in a cell-based screen for small molecule inhibitors of GLI1-mediated
n2:mentions: n3:19860666

Subject Item: _:vb12282616
rdf:type: n2:Context
rdf:value: GLI1 and GLI2 are the primary activators of HH signaling; further, the cooperative roles of GLI1 and GLI2 are critical in the transcriptional regulation of HH target genes[1, >>28<<-30]. While SMO has been extensively investigated as a therapeutic target[11-15, 17], few agents are available that target the GLI genes[31]. GANT61 was identified in a cell-based screen for small molecule inhibitors of GLI1-mediated
n2:mentions: n3:21317452 n3:12361967 n3:15175043

Subject Item: _:vb12282617
rdf:type: n2:Context
rdf:value: While SMO has been extensively investigated as a therapeutic target[>>11<<-15, 17], few agents are available that target the GLI genes[31].
n2:mentions: n3:20072843 n3:21300762 n3:19726763 n3:20412786

Subject Item: _:vb12282618
rdf:type: n2:Context
rdf:value: While SMO has been extensively investigated as a therapeutic target[11-15, >>17<<], few agents are available that target the GLI genes[31].
n2:mentions: n3:17353198

Subject Item: _:vb12282619
rdf:type: n2:Context
rdf:value: While SMO has been extensively investigated as a therapeutic target[11-15, 17], few agents are available that target the GLI genes[>>31<<]. GANT61 was identified in a cell-based screen for small molecule inhibitors of GLI1-mediated transcription. In the original study[31], GANT61 1) functions in the nucleus to abrogate GLI function; 2) blocks both GLI1- and GLI2- mediated
n2:mentions: n3:17494766

Subject Item: _:vb12282620
rdf:type: n2:Context
rdf:value: GANT61 was identified in a cell-based screen for small molecule inhibitors of GLI1-mediated transcription. In the original study[>>31<<], GANT61 1) functions in the nucleus to abrogate GLI function; 2) blocks both GLI1- and GLI2- mediated transcription; 3) in SUFU-null MEFS with constitutively active HH signaling, reduces expression of GLI1 and HIP1 mRNA in contrast to
n2:mentions: n3:17494766

Subject Item: _:vb12282621
rdf:type: n2:Context
rdf:value: We further demonstrated the specificity of GANT61 in targeting GLI1 and GLI2 from reduced GLI1 and GLI2 protein expression[>>8<<], inhibition of the binding of GLI1 and GLI2 to the promoter regions of HH target genes, specificity of reduction in GLI-luciferase reporter activity, and rapid inhibition of the transcriptional regulation of the GLI target gene BCL-2,
n2:mentions: n3:21135115

Subject Item: _:vb12282622
rdf:type: n5:Section
dc:title: inhibition of gli induces greater cytotoxicity than targeting smo
n5:contains: _:vb12282624 _:vb12282625 _:vb12282626 _:vb12282623

Subject Item: _:vb12282623
rdf:type: n2:Context
rdf:value: In time course studies, 24 hr exposure to GANT61 or 72 hr exposure to cyclopamine was required to initiate cell death[>>9<<]. In addition the SMO inhibitor GDC-0449, like cyclopamine, demonstrated limited cytotoxic activity in HT29 cells at equimolar concentrations (unpublished data). Collectively, the data demonstrate increased sensitivity of human colon
n2:mentions: n3:20399136

Subject Item: _:vb12282624
rdf:type: n2:Context
rdf:value: These concentrations and time frames for the induction of cellular effects are similar to those determined in other model systems for inhibitors of HH signaling[>>4<<, 19, 33].
n2:mentions: n3:20049737

Subject Item: _:vb12282625
rdf:type: n2:Context
rdf:value: These concentrations and time frames for the induction of cellular effects are similar to those determined in other model systems for inhibitors of HH signaling[4, >>19<<, 33].
n2:mentions: n3:19726788

Subject Item: _:vb12282626
rdf:type: n2:Context
rdf:value: These concentrations and time frames for the induction of cellular effects are similar to those determined in other model systems for inhibitors of HH signaling[4, 19, >>33<<].
n2:mentions: n3:20015350

Subject Item: _:vb12282627
rdf:type: n5:Section
dc:title: genetic downregulation of gli1 and gli2 by gli3r parallels cellular effects mediated by pharmacologic inhibition of gli1 and gli2 by gant61
n5:contains: _:vb12282632 _:vb12282633 _:vb12282634 _:vb12282635 _:vb12282636 _:vb12282628 _:vb12282629 _:vb12282630 _:vb12282631

Subject Item: _:vb12282628
rdf:type: n2:Context
rdf:value: A cleaved C-terminally truncated form of GLI3 (GLI3R) demonstrates repressor activity for GLI1 and GLI2 transcriptional regulation, and thereby silences HH-GLI target genes[>>4<<, 34]. We confirmed the critical role of GLI1 and GLI2 in cell survival by genetic downregulation of GLI1 and GLI2 following transient transfection of GLI3R[4, 34] in HT29 cells. Transient transfection and expression of GLI3R
n2:mentions: n3:20049737

Subject Item: _:vb12282629
rdf:type: n2:Context
rdf:value: A cleaved C-terminally truncated form of GLI3 (GLI3R) demonstrates repressor activity for GLI1 and GLI2 transcriptional regulation, and thereby silences HH-GLI target genes[4, >>34<<]. We confirmed the critical role of GLI1 and GLI2 in cell survival by genetic downregulation of GLI1 and GLI2 following transient transfection of GLI3R[4, 34] in HT29 cells. Transient transfection and expression of GLI3R (GLI3R-pCS2-MT,
n2:mentions: n3:10375510

Subject Item: _:vb12282630
rdf:type: n2:Context
rdf:value: We confirmed the critical role of GLI1 and GLI2 in cell survival by genetic downregulation of GLI1 and GLI2 following transient transfection of GLI3R[>>4<<, 34] in HT29 cells.
n2:mentions: n3:20049737

Subject Item: _:vb12282631
rdf:type: n2:Context
rdf:value: We confirmed the critical role of GLI1 and GLI2 in cell survival by genetic downregulation of GLI1 and GLI2 following transient transfection of GLI3R[4, >>34<<] in HT29 cells.
n2:mentions: n3:10375510

Subject Item: _:vb12282632
rdf:type: n2:Context
rdf:value: To begin to explore the cellular mechanisms downstream of GLI1/GLI2 inhibition that lead to cell death, we demonstrated that γH2AX, a marker of DNA DSBs[>>35<<], was expressed in GANT61-treated cells (Figure 4).
n2:mentions: n3:20139725

Subject Item: _:vb12282633
rdf:type: n2:Context
rdf:value: One of the critical regulators of DNA damage response is p53, which is stabilized upon DNA damage and modulates multiple components of the DNA damage response pathway[>>36<<, 37]. Expression of Gli3R-myc in human colon carcinoma cell lines harboring mutant p53 (functionally inactive) demonstrated DNA damage response suggesting a p53 independent mechanism[[32]]. Further, GANT61 treatment of HT29 cells
n2:mentions: n3:7475551

Subject Item: _:vb12282634
rdf:type: n2:Context
rdf:value: One of the critical regulators of DNA damage response is p53, which is stabilized upon DNA damage and modulates multiple components of the DNA damage response pathway[36, >>37<<]. Expression of Gli3R-myc in human colon carcinoma cell lines harboring mutant p53 (functionally inactive) demonstrated DNA damage response suggesting a p53 independent mechanism[[32]]. Further, GANT61 treatment of HT29 cells demonstrated
n2:mentions: n3:21293052

Subject Item: _:vb12282635
rdf:type: n2:Context
rdf:value: Figure 4Treatment of HT29 cells with vehicle alone (0.2% DMSO) or GANT61 (20 μM; upper panel), or transient transfection of vector alone, or GLI3R-pCS2-MT[>>4<<, 34](lower panel) for 72
n2:mentions: n3:20049737

Subject Item: _:vb12282636
rdf:type: n2:Context
rdf:value: Figure 4Treatment of HT29 cells with vehicle alone (0.2% DMSO) or GANT61 (20 μM; upper panel), or transient transfection of vector alone, or GLI3R-pCS2-MT[4, >>34<<](lower panel) for 72
n2:mentions: n3:10375510

Subject Item: _:vb12282637
rdf:type: n5:Section
dc:title: inhibition of gli1/gli2 by gant61 induces arrest in early s
n5:contains: _:vb12282640 _:vb12282641 _:vb12282638 _:vb12282639

Subject Item: _:vb12282638
rdf:type: n2:Context
rdf:value: In contrast 5-fluorouracil (FUra) combined with leucovorin (1 μM), which targets thymidylate synthase in the inhibition of DNA replication and induction of DNA damage, arrested HT29 cells in mid S-phase[>>38<<] (67.0% by FACS/PI staining; Figure 6A), thereby demonstrating the difference in S-phase target location of the mechanism of GANT61-induced inhibition of DNA replication.
n2:mentions: n3:11106264

Subject Item: _:vb12282639
rdf:type: n2:Context
rdf:value: The cell cycle checkpoints constitute a regulatory mechanism to arrest the cell cycle in response to DNA damage so that cell cycle progression and repair may be temporally coordinated[>>39<<-42]. In contrast to the checkpoints at the G1/S and G2/M transitions, the S-phase checkpoint can only delay the progression of S-phase. ATM is the master transducer of the S-phase checkpoint, responds to DNA DSBs, and together with its
n2:mentions: n3:15187248 n3:11850621 n3:2683079 n3:14744434

Subject Item: _:vb12282640
rdf:type: n2:Context
rdf:value: Activation of Cdk2 and the loading of Cdc45 onto replication origins are inhibited, DNA replication is inhibited, the intra-S-phase checkpoint is activated, and cyclin E accumulates[>>43<<-45], as demonstrated in GANT61-treated HT29 cells at 24 hr (Figure 6B).
n2:mentions: n3:12851482 n3:15000146 n3:15226429

Subject Item: _:vb12282641
rdf:type: n2:Context
rdf:value: Accumulation of HT29 cells in early S[>>9<<](Figure 6A), and the data of Figure 6B, suggest activation of an intra-S-phase checkpoint, which is not sustained.
n2:mentions: n3:20399136

Subject Item: _:vb470852920
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n2:RelevantScore: 11
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rdf:type: n2:RelevantBibliographicResource
n2:RelevantScore: 11
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rdf:type: n2:RelevantBibliographicResource
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rdf:type: n2:RelevantBibliographicResource
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n2:RelevantScore: 10
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rdf:type: n2:RelevantBibliographicResource
n2:RelevantScore: 9
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rdf:type: n2:RelevantBibliographicResource
n2:RelevantScore: 8
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rdf:type: n2:RelevantBibliographicResource
n2:RelevantScore: 8
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rdf:type: n2:RelevantBibliographicResource
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rdf:type: n2:RelevantBibliographicResource
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rdf:type: n2:RelevantBibliographicResource
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rdf:type: n2:RelevantBibliographicResource
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rdf:type: n2:RelevantBibliographicResource
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rdf:type: n2:RelevantBibliographicResource
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rdf:type: n2:RelevantBibliographicResource
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rdf:type: n2:RelevantBibliographicResource
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rdf:type: n2:RelevantBibliographicResource
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rdf:type: n2:RelevantBibliographicResource
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rdf:type: n2:RelevantBibliographicResource
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rdf:type: n2:RelevantBibliographicResource
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rdf:type: n2:RelevantBibliographicResource
n2:RelevantScore: 6
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rdf:type: n2:RelevantBibliographicResource
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rdf:type: n2:RelevantBibliographicResource
n2:RelevantScore: 5
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rdf:type: n2:RelevantBibliographicResource
n2:RelevantScore: 5
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rdf:type: n2:RelevantBibliographicResource
n2:RelevantScore: 5
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rdf:type: n2:RelevantBibliographicResource
n2:RelevantScore: 5
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rdf:type: n2:RelevantBibliographicResource
n2:RelevantScore: 5
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rdf:type: n2:RelevantBibliographicResource
n2:RelevantScore: 5
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rdf:type: n2:RelevantBibliographicResource
n2:RelevantScore: 5
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rdf:type: n2:RelevantBibliographicResource
n2:RelevantScore: 5
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rdf:type: n2:RelevantBibliographicResource
n2:RelevantScore: 5
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rdf:type: n2:RelevantBibliographicResource
n2:RelevantScore: 5
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