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Cited by 62Drug Transport and Resistance MechanismsCellular transport and secretionGlutathione Transferases and Polymorphisms

Glutathione-conjugate transport by RLIP76 is required for clathrin-dependent endocytosis and chemical carcinogenesis

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Singhal, Sharad S., Wickramarachchi, Dilki, Yadav, Sushma, Singhal, Jyotsana, Leake, Kathryn, Vatsyayan, Rit, Chaudhary, Pankaj, Lelsani, Poorna +4 more·2011·Molecular Cancer Therapeutics
DOI 10.1158/1535-7163.mct-10-0699PMC PMC3065778

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Abstract

Targeted depletion of the RALBP1-encoded 76-kDa splice variant, RLIP76, causes marked and sustained regression of human xenografts of lung, colon, prostate, and kidney cancers without toxicity in nude mouse models. We proposed that the remarkable efficacy and broad spectrum of RLIP76-targeted therapy is because its glutathione-conjugate (GS-E) transport activity is required for clathrin-dependent endocytosis (CDE), which regulates all ligand-receptor signaling, and that RLIP76 is required not only for survival of cancer cells but also for their very existence. We studied RLIP76 mutant proteins and the functional consequences of their expression into RLIP76−/− MEFs, identified key residues for GS-E binding in RLIP76, established the requirement of RLIP76-mediated GS-E transport for CDE, and showed a direct correlation between GS-E transport activities with CDE. Depletion of RLIP76 nearly completely blocked signaling downstream of EGF in a CDE-dependent manner and Wnt5a signaling in a CDE-independent manner. The seminal prediction of this hypothesis—RLIP76−/− mice will be deficient in chemical neoplasia—was confirmed. Benzo[a]pyrene, dimethylbenzanthracene, and phorbol esters are ineffective in causing neoplasia in RLIP76−/−. PMA-induced skin carcinogenesis in RLIP76+/+ mouse was suppressed completely by depletion of either PKCα or RLIP76 by siRNA or antisense and could be restored by topical application of RLIP76 protein in RLIP76−/− mouse skin. Likewise, chemical pulmonary carcinogenesis was absent in female and nearly absent in male RLIP76−/− mice. In RLIP76−/− mice, p53, p38, and JNK activation did not occur in response to either carcinogen. Our findings show a fundamental role of RLIP76 in chemical carcinogenesis. Mol Cancer Ther; 10(1); 16–28. ©2011 AACR.

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  • Rlip is a stress-protective ATPase of the mercapturic acid pathway that couples clathrin-dependent endocytosis with the efflux of glutathione-electrophile conjugates.

    “Rlip is a stress-protective ATPase of the mercapturic acid pathway that couples clathrin-dependent endocytosis with the efflux of glutathione–electrophile conjugates.”

    From Rlip76: An Unexplored Player in Neurodegeneration and Alzheimer’s Disease? (2022) · cites this paper as [3]

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Research works printed but not linked: 49

  1. [1]Awasthi S Singhal SS Sharma R Zimniak P Awasthi YC Transport of glutathione-conjugates and chemotherapeutic drugs by RLIP76: a novel link between G-protein and tyrosine-kinase signaling and drug-resistance Int J Cancer 2003 106 635 46 12866021
  2. [2]Awasthi S Singhal SS Srivastava SK Adenosine-triphosphate-dependent transport of doxorubicin, daunomyicn, and vinblastine in human tissues by a mechanism distinct from the P-glycoprotein J Clin Invest 1994 93 958 65 7907606 PMC294005
  3. [3]Huang Y Pharmacogenetics/genomics of membrane transporters in cancer chemotherapy Cancer Metastasis Rev 2007 26 183 201 17323126
  4. [4]Yadav S Zajac E Singhal SS Awasthi S Linking stress-signaling, glutathione-metabolism, signaling pathways and xenobiotic-transporters Cancer Metas Rev 2007 26 59 69 17260165
  5. [5]Desbois-Mouthon C Baron A Blivet-Van Eggelpoël MJ Insulin-like growth factor-1 receptor inhibition induces a resistance mechanism via the epidermal growth factor receptor/HER3/AKT signaling pathway: rational basis for cotargeting insulin-like growth factor-1 receptor and epidermal growth factor receptor in hepatocellular carcinoma Clin Cancer Res 2009 15 5445 56 19706799
  6. [6]Watabe T Miyazone K Roles of TGF-beta family signaling in stem cell renewal and differentiation Cell Res 2009 19 103 115 19114993
  7. [7]Guo Q Manolopoulou M Bian Y Schilling AB Tang WJ Molecular basis for the recognition and cleavages of IGF-II, TGF-alpha, and amylin by human insulin-degrading enzyme J Mol Biol 2010 395 430 43 19896952 PMC2813390
  8. [8]Baserga R Peruzzi F Reiss K The IGF-1receptor in cancer biology Int J Cancer 2003 107 873 7 14601044
  9. [9]Pollak M Insulin and insulin-like growth factor signaling in neoplasia Nat Rev Cancer 2008 8 915 28 19029956
  10. [10]Iozzo RV Eichstetter L Danielson KG Aberrant expression of the growth factor Wnt-5a in human malignancy Cancer Res 1995 55 3495 9 7627953
  11. [11]Laniosz V Holthusen KA Meneses PI Bovine papillomavirus type 1: from clathrin to caveolin J Virol 2008 82 6288 98 18417596 PMC2447075
  12. [12]Matsuzaki T Hanai S Kishi H Liu Z Bao Y Kikuchi A Regulation of endocytosis of activin type II receptors by a novel PDZ protein through Ral/Ral-binding protein 1-dependent pathway J Biol Chem 2002 277 19008 18 11882656
  13. [13]Zhang Y Zhang B TRAIL resistance of breast cancer cells is associated with constitutive endocytosis of death receptors 4 and 5 Mol Cancer Res 2008 6 1861 71 19074831
  14. [14]Pedersen NM Madshus IH Haslekås C Stang E Geldanamycin-induced down-regulation of ErbB2 from the plasma membrane is clathrin dependent but proteasomal activity independent Mol Cancer Res 2008 6 491 500 18337455
  15. [15]Rini BI Atkins MB Resistance to targeted therapy in renal-cell carcinoma Lancet Oncol 2009 10 992 1000 19796751
  16. [16]Jakoby WB The glutathione S-transferases: a group of multi-functional detoxification protein Adv Enzymol Mol Biol 1978 46 383 414 345769
  17. [17]Awasthi YC Sharma R Singhal SS Human glutathione S-transferases Int J Biochem 1994 26 295 308 8187927
  18. [18]Hayes JD Pulford DJ The glutathione S-transferase supergene family: regulation of GST and the contribution of the isoenzymes to cancer chemoprotection and drug resistance Crit Rev Biochem Mol Biol 1995 30 445 600 8770536
  19. [19]Awasthi S Cheng J Singhal SS Novel function of human RLIP76: ATP-dependent transport of glutathione-conjugates and doxorubicin Biochemistry 2000 39 9327 34 10924126
  20. [20]Stuckler D Singhal J Singhal SS Yadav S Awasthi YC Awasthi S RLIP76 transports vinorelbine and mediates drug-resistance in non-small cell lung cancer Cancer Res 2005 65 991 8 15705900
  21. [21]Singhal SS Yadav S Roth C Singhal J RLIP76: A novel glutathione-conjugate and multi-drug transporter Biochem Pharmacol 2009 77 761 9 18983828 PMC2664079
  22. [22]Singhal SS Yadav S Drake K Singhal J Awasthi S Hsf-1 and POB1 induce drug-sensitivity and apoptosis by inhibiting Ralbp1 J Biol Chem 2008 283 19714 29 18474607 PMC2443664
  23. [23]Quaroni A Paul EC Cytocentrin is a Ral-binding protein involved in the assembly and function of the mitotic-apparatus J Cell Sci 1999 112 707 18 9973605
  24. [24]Hu Y Mivechi NF HSF-1 interacts with Ral-binding-protein 1 in a stress-responsive, multi-protein complex with HSP90 in vivo J Biol Chem 2003 278 17299 306 12621024
  25. [25]Vatsyayan R Lelsani P Awasthi S Singhal SS RLIP76: A versatile transporter and an emerging target for cancer therapy Biochem Pharmacol 2010 79 1699 705 20097178 PMC2874070
  26. [26]Awasthi S Singhal SS Yadav S RALBP1 is a major determinant of radiation sensitivity Cancer Res 2005 65 6022 8 16024601
  27. [27]Singhal SS Yadav S Singhal J Awasthi YC Awasthi S Mitogenic and drug-resistance mediating effects of PKCα require RLIP76 Biochem Biophys Res Commun 2006 348 722 7 16890208 PMC2916682
  28. [28]Singhal J Singhal SS Yadav S RLIP76 in defense of radiation poisoning Int J Rad Oncol Biol Phys 2008 72 553 61 PMC2664086 18793957
  29. [29]Singhal SS Yadav S Singhal J Sahu M Sehrawat A Awasthi S Diminished drug transport and augmented radiation sensitivity caused by loss of RLIP76 FEBS Lett 2008 582 3408 14 18789326 PMC2586942
  30. [30]Singhal SS Wickramarachchi D Singhal J Yadav S Awasthi YC Awasthi S Determinants of differential doxorubicin sensitivity between SCLC and NSCLC FEBS Lett 2006 580 2258 64 16579994
  31. [31]Singhal SS Awasthi YC Awasthi S Regression of melanoma in a murine model by RLIP76-depletion Cancer Res 2006 66 2354 60 16489041
  32. [32]Singhal SS Singhal J Yadav S Regression of lung and colon cancer xenografts by depleting or inhibiting RLIP76 Cancer Res 2007 67 4382 9 17483352
  33. [33]Singhal SS Singhal J Yadav S Sahu M Awasthi YC Awasthi S RLIP76: A target for kidney cancer therapy Cancer Res 2009 69 4244 51 19417134 PMC2921230
  34. [34]Singhal SS Roth C Leake K Singhal J Yadav S Awasthi S Regression of prostate cancer xenografts by RLIP76 depletion Biochem Pharmacol 2009 77 1074 83 19073149 PMC2683040
  35. [35]Singhal SS Sehrawat A Sahu M RLIP76 transports sunitinib and sorafenib and mediates drug resistance in kidney cancer Int J Cancer 2010 126 1327 38 19626587 PMC3065777
  36. [36]Chan KS Sano S Kiguchi K Disruption of Stat3 reveals a critical role in both the initiation and the promotion stages of epithelial carcinogenesis J Clin Invest 2004 114 720 8 15343391 PMC514583
  37. [37]Chan KS Sano S Kataoka K Forced expression of a constitutively active form of Stat3 in mouse epidermis enhances malignant progression of skin tumors induced by two-stage carcinogenesis Oncogene 2008 27 1087 94 17700521
  38. [38]Rosse C L’Hoste S Offner N Picard A Camonis JH RLIP76, an effector of the Ral-GTPases, is a platform for Cdk1 to phosphorylate epsin during the switch off of endocytosis in mitosis J Biol Chem 2003 278 30597 604 12775724
  39. [39]Kariya K Koyama S Nakashima S Oshiro T Morinaka K Kikuchi A Regulation of complex formation of POB1/epsin/adaptor protein complex 2 by mitotic phosphorylation J Biol Chem 2000 275 18399 406 10764745
  40. [40]Hitchens TK Mannervik B Rule GS Disorder-to-order transition of the active site of human class Pi glutathione transferase, GST P1–1 Biochemistry 2001 40 11660 9 11570866
  41. [41]Chuang CC Wu SH Chiou SH Chang GG Homology modeling of cephalopod lens S-crystallin: a natural mutant of sigma-class glutathione transferase with diminished endogenous activity Biophys J 1999 76 679 90 9929473 PMC1300073
  42. [42]Tomarev SI Zinovieva RD Guo K Piatigorsky J Squid glutathione S-transferase. Relationships with other glutathione S-transferases and S-crystallins of cephalopods J Biol Chem 1993 268 4534 42 8440736
  43. [43]Awasthi S Cheng J Singhal SS Functional reassembly of xenobiotic transport from the N-terminal and C-terminal domains of RLIP76 and identification of ATP binding sequences Biochemistry 2001 40 4159 68 11300797
  44. [45]Singhal SS Yadav S Singhal J Drake K Awasthi YC Awasthi S The Role of PKCα and RLIP76 in Transport-mediated Doxorubicin-resistance in Lung Cancer FEBS Lett 2005 579 4635 41 16087181
  45. [46]Singhal SS Yadav S Singhal J Depletion of RLIP76 sensitizes lung cancer cells to doxorubicin Biochem Pharmacol 2005 70 481 8 15950949
  46. [47]Sharma R Haque AK Awasthi S Singh SV Piper JT Awasthi YC Differential carcinogenicity of benzo(a) pyrene in male and female CD-1 mouse lung J Toxicol Environ Health 1997 52 45 62 9269322
  47. [48]Awasthi S Singhal SS Yadav S A central role of RLIP76 in regulation of glycemic control Diabetes 2010 59 714 25 20007934 PMC2828645
  48. [49]Cheng JZ Sharma R Yang Y Accelerated metabolism and exclusion of 4-hydroxynonenal through induction of RLIP76 and hGST5.8 is an early adaptive response of cells to heat and oxidative stress J Biol Chem 2001 276 41213 23 11522795
  49. [50]Michie AM Nakagawa R The link between PKCα regulation and cellular transformation Immunol Lett 2005 96 155 62 15585319

Cited by

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Authors
Singhal, S. S., Wickramarachchi, D., Yadav, S., Singhal, J., Leake, K., Vatsyayan, R., Chaudhary, P., Lelsani, P., Suzuki, S., Yang, S., Awasthi, Y. C., & Awasthi, S.
Affiliations
  • Scripps Research Institute (US)
  • Authors' Affiliations: 1Department of Molecular Biology and Immunology, University of North Texas Health Science Center, Fort Worth, Texas; 2Immunology and Microbiology, The Scripps Research Institute, La Jolla, California; and Departments of 3Biostatistics, School of Public Health, and 4Pharmacology and Neuroscience, University of North Texas Health Science Center, Fort Worth, Texas
Corresponding author
Singhal, Sharad S.
ORCID
Venue
Molecular Cancer Therapeutics
Publisher
American Association for Cancer Research (AACR)
Published
2011-01-01
Volume
10
Issue
1
Pages
16-28
ISSN
  • 1535-7163 (ISSN-L)
  • 1538-8514
DOI
10.1158/1535-7163.mct-10-0699
PMC
PMC3065778
OpenAlex
W2133735016
Cited by
62
References
0
Citations by year
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FWCI
1.93
Type
journal article
Language
en
Open access
Locations
  • Molecular Cancer Therapeutics (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • National Cancer Institute (CA 77495, R01CA104661, R01CA077495, CA104661)
  • National Institute of Environmental Health Sciences (ES012171, R01ES012171)
Topics
  • Health Sciences › Medicine › Oncology › Drug Transport and Resistance Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glutathione Transferases and Polymorphisms
Keywords
RLIP76, glutathione conjugate transport, clathrin-mediated endocytosis, chemical carcinogenesis, skin carcinogenesis, pulmonary carcinogenesis, EGF signaling, Wnt5a signaling, benzo[a]pyrene, phorbol ester, protein kinase C alpha, p53 activation, p38 MAPK, JNK, tumor regression, xenografts
MeSH terms
Animals, Binding Sites, Carcinogens, Clathrin, Endocytosis, Female, Glutathione, Humans, Male, Skin Neoplasms, Tetradecanoylphorbol Acetate, Transfection, 9,10-Dimethyl-1,2-benzanthracene, Signal Transduction, Tumor Suppressor Protein p53, Mice, Knockout, Drug Resistance, Neoplasm, GTPase-Activating Proteins, Mice, Protein Kinase C-alpha

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