Characterization of HCT116 Human Colon Cancer Cells in an Orthotopic Model

Rajput, Ashwani; Martin, Ivan; Rose, Rebecca; Beko, Alexander; LeVea, Charles; Sharratt, Elizabeth; Mazurchuk, Richard; Hoffman, Robert M.; Brattain, Michael G.; Wang, Jing

doi:10.1016/j.jss.2007.04.021

Cited by 132 publications

(111 citation statements)

References 25 publications

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“…Experimental protocols were approved by The Johns Hopkins University Animal Care and Use Committee. The xenograft and orthotopic models were performed as described (26).…”

Section: Methodsmentioning

confidence: 99%

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Inositol pyrophosphates promote tumor growth and metastasis by antagonizing liver kinase B1

Rao

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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The inositol pyrophosphates, molecular messengers containing an energetic pyrophosphate bond, impact a wide range of biologic processes. They are generated primarily by a family of three inositol hexakisphosphate kinases (IP6Ks), the principal product of which is diphosphoinositol pentakisphosphate (IP7). We report that IP6K2, via IP7 synthesis, is a major mediator of cancer cell migration and tumor metastasis in cell culture and in intact mice. IP6K2 acts by enhancing cell-matrix adhesion and decreasing cell-cell adhesion. This action is mediated by IP7-elicited nuclear sequestration and inactivation of the tumor suppressor liver kinase B1 (LKB1). Accordingly, inhibitors of IP6K2 offer promise in cancer therapy.I nositol pyrophosphates, conserved eukaryotic messenger molecules with a pyrophosphate bond, mediate numerous physiologic processes including regulation of Akt (1), insulin secretion (2), ATP production (3), DNA repair (4), and damage response (5). Exemplified by diphosphoinositol pentakisphosphate (PPIP5, IP7), inositol pyrophosphates are primarily generated by a family of three inositol hexakisphosphate (IP6) kinases (IP6K) (6, 7) and also may be formed by a more recently described group of IP6/7 kinases (8). IP6K1 and IP6K2 are widely distributed, whereas IP6K3 is expressed primarily in the brain (9). A related enzyme, inositol polyphosphate multikinase (IPMK), converts IP3 to IP4 and IP5, displays physiologic PI3 kinase activity (10), and noncatalytically acts as a transcriptional coactivator (11) and stabilizer of the mTOR complex-1 (12). Whether IP6Ks and IPMK play direct roles in tumor progression remains unexplored.Tumor cell metastasis requires loss of cell-cell adhesion and gain of migratory and invasive properties, a collective program known as epithelial-mesenchymal transition (EMT), which is also critical for embryonic development (13). Cell-cell adhesion and cell migration/invasion are primarily driven by E-cadherinmediated cell clustering (14) and focal-adhesion-based cellmatrix interactions (15), respectively. Whether these two distinct adhesion processes are concurrently or independently regulated remains poorly understood.The tumor suppressor LKB1 has been separately reported to inhibit FAK activation (16) and to enhance E-cadherin expression (17)(18)(19). LKB1 is mutated in Peutz-Jeghers syndrome patients who tend to develop cancer at multiple sites (20). As a master kinase controlling the activity of AMPK and 13 other AMPK-like kinases (21), LKB1 mediates diverse cellular processes such as polarity, adhesion, metabolism, tumor growth/metastasis, and neuronal axon initiation/branching (22). Unlike conventional protein kinases, LKB1 is not activated by activation loop phosphorylation but by protein-protein interaction with two other subunits of the heterotrimeric holoenzyme: STRAD and Mo25. LKB1 can also exist as an inactive nuclear monomer. STRAD binds and stabilizes LKB1 (23) in the cytosol, where it is phosphorylated at serine 428 by PKCζ (24). Whether cytosolic localization enhance...

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“…Experimental protocols were approved by The Johns Hopkins University Animal Care and Use Committee. The xenograft and orthotopic models were performed as described (26).…”

Section: Methodsmentioning

confidence: 99%

“…2E), consistent with a role for IP6K2 in tumor growth. To study the influence of IP6K2 on tumor metastasis, we used an established orthotopic model wherein small pieces of xenograft tumors are implanted into the cecum of nude mice (26) and liver metastases are monitored (Fig. 2 F and G).…”

Section: Significancementioning

confidence: 99%

Inositol pyrophosphates promote tumor growth and metastasis by antagonizing liver kinase B1

Rao

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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“…To investigate the roles of miR-103 and miR-107 in CRC metastasis, we examined their expression in a panel of CRC cell lines showing differential metastasis potential in an orthotopic model (26)(27)(28)(29)(30). Using qPCR analysis with primers specific to miR-103 and miR-107 ( Supplementary Fig.…”

Section: Mir-103/107 Expression Correlates With Crc Metastasis and Pomentioning

confidence: 99%

miR-103/107 Promote Metastasis of Colorectal Cancer by Targeting the Metastasis Suppressors DAPK and KLF4

et al. 2012

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Metastasis is the major cause of poor prognosis in colorectal cancer (CRC), and increasing evidence supports the contribution of miRNAs to cancer progression. Here, we found that high expression of miR-103 and miR-107 (miR-103/107) was associated with metastasis potential of CRC cell lines and poor prognosis in patients with CRC. We showed that miR-103/107 targeted the known metastasis suppressors death-associated protein kinase (DAPK) and Kr€ uppel-like factor 4 (KLF4) in CRC cells, resulting in increased cell motility and cell-matrix adhesion and decreased cell-cell adhesion and epithelial marker expression. miR-103/107 expression was increased in the presence of hypoxia, thereby potentiating DAPK and KLF4 downregulation and hypoxiainduced motility and invasiveness. In mouse models of CRC, miR-103/107 overexpression potentiated local invasion and liver metastasis effects, which were suppressed by reexpression of DAPK or KLF4. miR-103/107-mediated downregulation of DAPK and KLF4 also enabled the colonization of CRC cells at a metastatic site. Clinically, the signature of a miR-103/107 high, DAPK low, and KLF4 low expression profile correlated with the extent of lymph node and distant metastasis in patients with CRC and served as a prognostic marker for metastasis recurrence and poor survival. Our findings therefore indicate that miR-103/107-mediated repression of DAPK and KLF4 promotes metastasis in CRC, and this regulatory circuit may contribute in part to hypoxia-stimulated tumor metastasis. Strategies that disrupt this regulation might be developed to block CRC metastasis. Cancer Res; 72(14); 3631-41. Ó2012 AACR.

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“…In the present study, to investigate coordination between energy metabolism and cell-cycle progression of cancer cells in vivo, we have used a human colorectal cancer cell line HCT116 to provide a liver metastasis model in superimmunodeficient NOD/SCID/IL-2Rg c null (NOG) mice (14)(15)(16)(17)(18). To examine energy metabolism of individual cancer cells in metastatic tumors, we used HCT116 cells labeled with a cell-cycle phase-dependent fluorescent marker system Fucci (19).…”

Section: Introductionmentioning

confidence: 99%

Energy Management by Enhanced Glycolysis in G1-phase in Human Colon Cancer Cells In Vitro and In Vivo

Bao

Mukai

Hishiki

et al. 2013

Molecular Cancer Research

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Activation of aerobic glycolysis in cancer cells is well known as the Warburg effect, although its relation to cellcycle progression remains unknown. In this study, human colon cancer cells were labeled with a cell-cycle phasedependent fluorescent marker Fucci to distinguish cells in G 1 -phase and those in S þ G 2 /M phases. Fucci-labeled cells served as splenic xenograft transplants in super-immunodeficient NOG mice and exhibited multiple metastases in the livers, frozen sections of which were analyzed by semiquantitative microscopic imaging mass spectrometry. Results showed that cells in G 1 -phase exhibited higher concentrations of ATP, NADH, and UDP-N-acetylglucosamine than those in S and G 2 -M phases, suggesting accelerated glycolysis in G 1 -phase cells in vivo. Quantitative determination of metabolites in cells synchronized in S, G 2 -M, and G 1 phases suggested that efflux of lactate was elevated significantly in G 1 -phase. By contrast, ATP production in G 2 -M was highly dependent on mitochondrial respiration, whereas cells in S-phase mostly exhibited an intermediary energy metabolism between G 1 and G 2 -M phases. Isogenic cells carrying a p53-null mutation appeared more active in glycolysis throughout the cell cycle than wild-type cells. Thus, as the cell cycle progressed from G 2 -M to G 1 phases, the dependency of energy production on glycolysis was increased while the mitochondrial energy production was reciprocally decreased.Implications: These results shed light on distinct features of the phase-specific phenotypes of metabolic systems in cancer cells. Mol Cancer Res; 11(9); 973-85. Ó2013 AACR.

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Characterization of HCT116 Human Colon Cancer Cells in an Orthotopic Model

Cited by 132 publications

References 25 publications

Inositol pyrophosphates promote tumor growth and metastasis by antagonizing liver kinase B1

Inositol pyrophosphates promote tumor growth and metastasis by antagonizing liver kinase B1

miR-103/107 Promote Metastasis of Colorectal Cancer by Targeting the Metastasis Suppressors DAPK and KLF4

Energy Management by Enhanced Glycolysis in G1-phase in Human Colon Cancer Cells In Vitro and In Vivo

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