scholarly journals Heterogeneity of Metabolic Vulnerability in Imatinib-Resistant Gastrointestinal Stromal Tumor

Cells ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 1333
Author(s):  
Wen-Kuan Huang ◽  
Jiwei Gao ◽  
Ziqing Chen ◽  
Hao Shi ◽  
Juan Yuan ◽  
...  
Keyword(s):  
Cell Lines ◽  
Mitochondrial Respiration ◽  
Resistant Cell ◽  
Resistant Cell Line ◽  
Metabolic Reprogramming ◽  
Peroxisome Proliferator ◽  
Imatinib Treatment ◽  
Peroxisome Proliferator Activated Receptor ◽  
Glycolytic Activity ◽  
Imatinib Resistant

Metabolic reprogramming is a hallmark of cancer cells in response to targeted therapy. Decreased glycolytic activity with enhanced mitochondrial respiration secondary to imatinib has been shown in imatinib-sensitive gastrointestional stromal tumors (GIST). However, the role of energy metabolism in imatinib-resistant GIST remains poorly characterized. Here, we investigated the effect of imatinib treatment on glycolysis and oxidative phosphorylation (OXPHOS), as well as the effect of inhibition of these energy metabolisms on cell viability in imatinib-resistant and -sensitive GIST cell lines. We observed that imatinib treatment increased OXPHOS in imatinib-sensitive, but not imatinib-resistant, GIST cells. Imatinib also reduced the expression of mitochondrial biogenesis activators (peroxisome proliferator-activated receptor coactivator-1 alpha (PGC1α), nuclear respiratory factor 2 (NRF2), and mitochondrial transcription factor A (TFAM)) and mitochondrial mass in imatinib-sensitive GIST cells. Lower TFAM levels were also observed in imatinib-sensitive GISTs than in tumors from untreated patients. Using the Seahorse system, we observed bioenergetics diversity among the GIST cell lines. One of the acquired resistant cell lines (GIST 882R) displayed a highly metabolically active phenotype with higher glycolysis and OXPHOS levels compared with the parental GIST 882, while the other resistant cell line (GIST T1R) had a similar basal glycolytic activity but lower mitochondrial respiration than the parental GIST T1. Further functional assays demonstrated that GIST 882R was more vulnerable to glycolysis inhibition than GIST 882, while GIST T1R was more resistant to OXPHOS inhibition than GIST T1. These findings highlight the diverse energy metabolic adaptations in GIST cells that allow them to survive upon imatinib treatment and reveal the potential of targeting the metabolism for GIST therapy.

The Proteome Analysis of an Imatinib-Resistant Cell Line Identifies Changes in Molecular Chaperones Such as Heat Shock Proteins (Hsp): A Possible New Mechanism of Imatinib Resistance.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 1981-1981 ◽  
Author(s):  
Jean-Max Pasquet ◽  
Marion Pocaly ◽  
Valérie Lagarde ◽  
Béatrice Turcq ◽  
Josy Reiffers ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Heat Shock ◽  
Cell Line ◽  
Cell Lines ◽  
Molecular Chaperones ◽  
Resistant Cell ◽  
Resistant Cell Line ◽  
Mechanism Of Resistance ◽  
Resistant Phenotype ◽  
Imatinib Resistant

Abstract Targeting the tyrosine kinase activity of Bcr-Abl by imatinib mesylate is an attractive therapeutic strategy in chronic myelogenous leukemia (CML) and in Bcr-Abl positive acute lymphoblastic leukemia. However, resistance to imatinib monotherapy is currently a major issue preventing the successful treatment of CML patients. It may be mainly mediated by mutations within the kinase domain of Bcr-Abl and/or amplification of the BCR-ABL genomic locus. The K562-r imatinib-resistant cell line, derived in our laboratories from the sensitive parental K562-s has neither mechanism of resistance, nor overexpression of Src-kinases such as Lyn and Hck, as described for other cell lines. In the current study we used two-dimensional (2D) difference gel electrophoresis (DIGE) and MALDI-TOF-TOF mass spectrometry to compare the proteome of K562-r and K562-s. With the aid of the Image Master TM 2D platinium software, we detected 31 different proteins in K562-r and K562-s. These proteins were classified in 3 different groups. The first includes proteins involved in the synthesis and stability of RNA (hnRNP K, hnRNP H, CstF , transcription elongation factor A protein 1, PCBP2, TCP1), the second encompasses structural proteins (CAPG, fascin, tubulin, vimentin, laminA, C tubulin beta-1 chain, actin cytoplasmic 1, keratin type I type II), and the third was represented by different enzymes participating in general metabolic pathways (glyceraldehyde 3-phosphate dehydrogenase, malate dehydrogenase, mitochondrial precursor, glutamate dehydrogenase 2, pyruvate kinase, PURH protein). Furthermore, chaperone proteins such as heat-shock protein Hsp60, P60HOP or STI-1, Hsp105 and Hsp70 were differentially expressed in the sensitive and resistant cell lines. Since these proteins complex with Hsp90 and this complex has been reported to interact with the Bcr-Abl protein, we focused on these molecular chaperones. Hsp70 family proteins such as Hsc70 and Hsp74 were found to be more expressed 2.5-fold higher in K562-r than in K562-s, and/or exhibited post translational modifications (phosphorylation and acetylation) confirmed by Western blotting. Hsp70 was recently described as an inhibitor of apoptosis (Ray S et al., JBC 2004) and its overexpression in K562-r could thus contribute to its imatinib-resistant phenotype. Preliminary functional studies showed that whereas K562-s and K562-r were equally sensitive to the apoptotic effect of geldanamycin (an inhibitor of Hsp90), the combination of geldanamycin and a proteasome inhibitor (MG132) was more efficient in K562-r than in K562-s (viability of 16% and 40% respectively after 4 days in culture). Ongoing experiments utilizing siRNA against Hsp70 will help understand the link between the expression profile of Hsp proteins and the imatinib-resistant phenotype of this cell line. In conclusion, the use of a new experimental strategy, i.e. proteomic analysis by DIGE and mass spectrometry, allowed us to identify selected proteins whose patterns of expression and post-translational modification may underlie a new mechanism of resistance to imatinib in Bcr-Abl positive cells.

A novel natural compound, a cycloanthranilylproline derivative (Fuligocandin B), sensitizes leukemia cells to apoptosis induced by tumor necrosis factor–related apoptosis-inducing ligand (TRAIL) through 15-deoxy-Δ12, 14 prostaglandin J2 production

Blood ◽  
2007 ◽  
Vol 110 (5) ◽  
pp. 1664-1674 ◽  
Author(s):  
Hiroo Hasegawa ◽  
Yasuaki Yamada ◽  
Kanki Komiyama ◽  
Masahiko Hayashi ◽  
Masami Ishibashi ◽  
...  
Keyword(s):  
Tumor Necrosis Factor ◽  
Tumor Necrosis ◽  
Leukemia Cell ◽  
Resistant Cell ◽  
Resistant Cell Line ◽  
Peroxisome Proliferator ◽  
Independent Manner ◽  
Peroxisome Proliferator Activated Receptor ◽  
Necrosis Factor ◽  
Prostaglandin J2

Abstract Tumor necrosis factor (TNF)–related apoptosis-inducing ligand (TRAIL) induces apoptosis in many transformed cells; however, not all human tumors respond to TRAIL, potentially limiting its therapeutic utility. Although there is substantial evidence that cytotoxic drugs can augment sensitivity to TRAIL, it has become important to know what kinds of nontoxic drugs can be used together with TRAIL. We thus screened several natural compounds that can overcome resistance to TRAIL and found that a cycloanthranilylproline derivative, Fuligocandin B (FCB), an extract of myxomycete Fuligo candida, exhibited significant synergism with TRAIL. Treatment of the TRAIL-resistant cell line KOB with FCB and TRAIL resulted in apparent apoptosis, which was not induced by either agent alone. FCB increased the production of 15-deoxy-Δ12,14 prostaglandin J2 (15d-PGJ2), an endogenous PPARγ ligand, through activation of cyclooxygenase-2 (COX-2). This unique mechanism highlighted the fact that 15d-PGJ2 directly enhanced sensitivity to TRAIL by inhibiting multiple antiapoptotic factors. More importantly, similar effects were observed in other leukemia cell lines irrespective of their origin. The enhancement was observed regardless of PPARγ expression and was not blocked even by peroxisome proliferator-activated receptor-γ (PPARγ) siRNA. These results indicate that 15d-PGJ2 sensitizes TRAIL-resistant cells to TRAIL in a PPARγ-independent manner and that the use of 15d-PGJ2 or its inducers, such as FCB, is a new strategy for cancer therapy.

scholarly journals Growth Hormone Inhibits Apoptosis in Human Colonic Cancer Cell Lines: Antagonistic Effects of Peroxisome Proliferator Activated Receptor-γ Ligands

Endocrinology ◽  
2004 ◽  
Vol 145 (7) ◽  
pp. 3353-3362 ◽  
Author(s):  
Fausto Bogazzi ◽  
Federica Ultimieri ◽  
Francesco Raggi ◽  
Dania Russo ◽  
Renato Vanacore ◽  
...  
Keyword(s):  
Cell Lines ◽  
Cancer Cell ◽  
Colonic Cancer ◽  
Cancer Cell Lines ◽  
Peroxisome Proliferator ◽  
Signal Transducer ◽  
Peroxisome Proliferator Activated Receptor ◽  
Antiapoptotic Effect ◽  
Pparγ Expression ◽  
Colonic Cells

Abstract GH has antiapoptotic effects on several cells. However, the antiapoptotic mechanisms of GH on colonic mucosa cells are not completely understood. Peroxisome proliferator activated receptor-γ (PPARγ) activation enhances apoptosis, and a link between GH and PPARγ in the colonic epithelium of acromegalic patients has been suggested. We investigated the effects of GH and of PPARγ ligands on apoptosis in colonic cancer cell lines. Colonic cells showed specific binding sites for GH, and after exposure to 0.05–50 nm GH, their apoptosis reduced by 45%. The antiapoptotic effect was due to either GH directly or GH-dependent local production of IGF-1. A 55–85% reduction of PPARγ expression was observed in GH-treated cells, compared with controls (P < 0.05). However, treatment of the cells with 1–50 μm ciglitazone (cig), induced apoptosis and reverted the antiapoptotic effects of GH by increasing the programmed cell death up to 3.5-fold at 30 min and up to 1.7-fold at 24 h. Expression of Bcl-2 and TNF-related apoptosis-induced ligand was not affected by either GH or cig treatment, whereas GH reduced the expression of Bax, which was increased by cig treatment. In addition, GH increased the expression of signal transducer and activator of transcription 5b, which might be involved in the down-regulation of PPARγ expression. In conclusion, GH may exert a direct antiapoptotic effect on colonic cells, through an increased expression of signal transducer and activator of transcription 5b and a reduction of Bax and PPARγ. The reduced GH-dependent apoptosis can be overcome by PPARγ ligands, which might be useful chemopreventive agents in acromegalic patients, who have an increased colonic polyps prevalence.

Avian Sarcoma Virus Transformed Hamster Cells made Resistant to Ethidium Bromide

1974 ◽  
Vol 16 (3) ◽  
pp. 603-621
Author(s):  
C. ALTANER ◽  
J. MATOSKA
Keyword(s):  
Cell Lines ◽  
Ethidium Bromide ◽  
Virus Genome ◽  
Resistant Cell ◽  
Resistant Cell Line ◽  
Avian Sarcoma Virus ◽  
Sarcoma Virus ◽  
Original Cell ◽  
Avian Sarcoma ◽  
Resistant Cells

Hamster cells transformed with the Schmidt-Ruppin strain of avian sarcoma virus were selected for resistance to ethidium bromide (EB). The resistant cell lines proliferated in the presence of up to 30 µg/ml EB. From avian sarcoma virus-transformed hamster cells already resistant to bromodeoxy-uridine (BrdU), ethidium bromide-resistant cells which were able to grow in 10 µg/ml EB were also prepared. These cells remain deficient in thymidine kinase activity and are suitable for selective preparation of hybrid cells. The EB resistance was genetically stable. The EB-resistant cell lines, and doubly resistant cells (BrdU, EB) showed no differences in mitochondrial ultrastructure compared with the original cell lines. Thymidine incorporation into mitochondrial DNA was not influenced by EB resistance. All resistant cell lines, including the doubly resistant cell line, contained the avian sarcoma virus genome. The number of cells needed for positive rescue experiments for avian sarcoma virus genome by cell fusion with permissive chicken embryo cells was the same as with the original cell lines. The single EB-resistant cell lines contained R-type virus-like particles, while in BrdU-resistant and doubly resistant cells the R-type particles were absent. The possible nature of EB resistance is discussed.

scholarly journals The Peroxisome Proliferator-Activated Receptor Gamma System Regulates Ultraviolet B-Induced ProstaglandinE2Production in Human Epidermal Keratinocytes

PPAR Research ◽  
2010 ◽  
Vol 2010 ◽  
pp. 1-8 ◽  
Author(s):  
Raymond L. Konger ◽  
Kellie Clay Martel ◽  
Danielle Jernigan ◽  
Qiwei Zhang ◽  
Jeffrey B. Travers
Keyword(s):  
Cell Lines ◽  
Mouse Skin ◽  
Ultraviolet B ◽  
Human Epidermis ◽  
Epidermal Keratinocytes ◽  
Peroxisome Proliferator ◽  
Primary Human Keratinocytes ◽  
Peroxisome Proliferator Activated Receptor ◽  
Potent Inducer ◽  
Cox 2

Studies using PPARγagonists in mouse skin have suggested that peroxisome proliferator-activated receptor gamma (PPARγ) is irrelevant to cutaneous photobiology. However, in several epithelial cell lines, ultraviolet B (UVB) has been shown to induce the nonenzymatic production of oxidized phospholipids that act as PPARγagonists. UVB is also a potent inducer of prostaglandinE2  (PGE2)production and COX-2 expression in keratinocytes and PPARγis coupled to increasedPGE2production in other cell lines. In this current study, we demonstrate that PPARγagonists, but not PPARαor PPARβ/δagonists, inducePGE2production and COX-2 expression in primary human keratinocytes (PHKs). Importantly, PPARγagonist-induced COX-2 expression andPGE2production were partially inhibited by the PPARγantagonist, GW9662, indicating that both PPARγ-dependent and -independent pathways are likely involved. GW9662 also suppressed UVB andtert-butylhydroperoxide- (TBH-) inducedPGE2production in PHKs and intact human epidermis and partially inhibited UVB-induced COX-2 expression in PHKs. These findings provide evidence that PPARγis relevant to cutaneous photobiology in human epidermis.

Peroxisome proliferator-activated receptor ? in the human breast cancer cell lines MCF-7 and MDA-MB-231

2002 ◽  
Vol 34 (4) ◽  
pp. 165-171 ◽  
Author(s):  
Kate M. Suchanek ◽  
Fiona J. May ◽  
Jodie A. Robinson ◽  
Won Jae Lee ◽  
Nicola A. Holman ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Lines ◽  
Breast Cancer Cell ◽  
Human Breast Cancer ◽  
Human Breast Cancer Cell ◽  
Breast Cancer Cell Lines ◽  
Peroxisome Proliferator ◽  
Human Breast ◽  
Peroxisome Proliferator Activated Receptor ◽  
Mcf 7
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