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01-08-2013 | Research Article

Effects of Rab27a on proliferation, invasion, and anti-apoptosis in human glioma cell

Authors: Xiuwei Wu, Anla Hu, Mingjun Zhang, Zhendong Chen

Published in: Tumor Biology | Issue 4/2013

Abstract

This study aims to investigate the relationship between Rab27a and the characteristics of glioma cell U251 such as proliferation, apoptosis, and invasion and to provide an experimental basis for future therapy in human glioma. Recombinant plasmid of pcDNA3.1-Rab27a was constructed and transfected into U251 cells with the help of Lipofectamine™2000. The expression of Rab27a was detected by Western blot. Cell viability, cell cycle, cell apoptosis, and cell migration were analyzed, respectively, by (3-(4,5)-dimethylthi-azol-2-yl)-2,5-diphenytetrazolium bromide (MTT) assay, flow cytometry, and Transwell invasion chamber methods. Meanwhile, the effect of Rab27a on secretion of cathepsin D in U251 cells was also examined. With the help of luciferase reporter assay system, the relationship between miR-124 and gene Rab27a expression was explored. Western blot showed that the expression of Rab27a was significantly increased in pcDNA3.1-Rab27a transfection group (p < 0.01) and that was significantly decreased in Rab27a-shRNA transfection group (p < 0.01) compared with control group. MTT assay, flow cytometry, and Transwell invasion chamber experiment indicated that cell viability (p < 0.01), proliferation index (p < 0.05), and invasion ability (p < 0.01) were improved significantly in pcDNA3.1-Rab27a transfection group compared with control group and that cell viability (p < 0.01), proliferation index (p < 0.05), and invasion ability (p < 0.01) were reduced markedly in Rab27a-shRNA transfection group compared with control group. The apoptosis analysis by flow cytometry demonstrated that the ratio of apoptosis in pcDNA3.1-Rab27a transfection group was significantly lower than that in control group (p < 0.05) and the ratio was notably higher in Rab27a-shRNAtransfection group than that in the control group. Cathepsin D activity assay indicated that the release of cathepsin D was enhanced in pcDNA3.1-Rab27a transfection group compared to that in the control group (p < 0.05). Rab27a could increase the glioma cell ability, promote proliferation and invasion, and suppress cell apoptosis. The above-stated effects of Rab27a possibly were exerted by increasing the secretion of cathepsin D and regulated by miR-124. In addition, the inhibition of expression of Rab27a perhaps benefited the therapy for glioma patients.

Shah AH, Snelling B, Bregy A, Patel PR, Tememe D, Bhatia R, et al. Discriminating radiation necrosis from tumor progression in gliomas: a systematic review what is the best imaging modality? J Neurooncol. 2013. doi:10.1007/s11060-013-1059-9.

Chen S, Tanaka S, Giannini C, Morris J, Yan ES, Buckner J, et al. J Neurooncol. 2013. doi:10.1007/s11060-013-1058-x.

van Horssen R, Willemse M, Haeger A, Attanasio F, Guneri T, Schwab A, et al. Intracellular NAD(H) levels control motility and invasion of glioma cells. Cell Mol Life Sci. 2013. doi:10.1007/s00018-012-1249-1.PubMed

Mao F, Wang B, Xiao Q, Xi G, Sun W, Zhang H, et al. A role for LRIG1 in the regulation of malignant glioma aggressiveness. Int J Oncol. 2013;42(3):1081–7.PubMed

Hdeib A, Sloan AE. Convection-enhanced delivery of (131)I-chTNT-1/B mAB for treatment of high-grade adult gliomas. Expert Opin Biol Ther. 2011;11(6):799–806.PubMedCrossRef

Strik HM, Kolodziej M, Oertel W, Basecke J. Glycobiology in malignant gliomas: expression and functions of galectins and possible therapeutic options. Curr Pharm Biotechnol. 2012;13(11):2299–307.PubMedCrossRef

Stenmark H. Rab GTPases as coordinators of vesicle traffic. Nat Rev Mol Cell Biol. 2009;10(8):513–25.PubMedCrossRef

Svetina S. Vesicle budding and the origin of cellular life. ChemPhysChem. 2009;10(16):2769–76.PubMedCrossRef

Barth J, Volknandt W. Proteomic investigations of the synaptic vesicle interactome. Expert Rev Proteomics. 2011;8(2):211–20.PubMedCrossRef

10.

Kelly BT, Owen DJ. Endocytic sorting of transmembrane protein cargo. Curr Opin Cell Biol. 2011;23(4):404–12.PubMedCrossRef

11.

Neto H, Collins LL, Gould GW. Vesicle trafficking and membrane remodelling in cytokinesis. Biochem J. 2011;437(1):13–24.PubMedCrossRef

12.

Hutagalung AH, Novick PJ. Role of Rab GTPases in membrane traffic and cell physiology. Physiol Rev. 2011;91(1):119–49.PubMedCrossRef

13.

Wang T, Ming Z, Xiaochun W, Hong W. Rab7: role of its protein interaction cascades in endo-lysosomal traffic. Cell Signal. 2011;23(3):516–21.PubMedCrossRef

14.

Angers CG, Merz AJ. New links between vesicle coats and Rab-mediated vesicle targeting. Semin Cell Dev Biol. 2011;22(1):18–26.PubMedCrossRef

15.

Brighouse A, Dacks JB, Field MC. Rab protein evolution and the history of the eukaryotic endomembrane system. Cell Mol Life Sci. 2011;67(20):3449–65.CrossRef

16.

Recchi C, Seabra MC. Novel functions for Rab GTPases in multiple aspects of tumour progression. Biochem Soc Trans. 2012;40(6):1398–403.PubMedCrossRef

17.

Musilli M, Nicolia V, Borrelli S, Scarpa S, Diana G. Behavioral effects of Rho GTPase modulation in a model of Alzheimer’s disease. Behav Brain Res. 2013;237:223–9.PubMedCrossRef

18.

Yoshida-Amano Y, Hachiya A, Ohuchi A, Kobinger GP, Kitahara T, Takema Y, et al. Essential role of RAB27A in determining constitutive human skin color. PLoS One. 2012;7(7):e41160.PubMedCrossRef

19.

Arora DK, Syed I, Machhadieh B, McKenna CE, Kowluru A. Rab-geranylgeranyl transferase regulates glucose-stimulated insulin secretion from pancreatic beta cells. Islets. 2012;4(5):354–8.PubMedCrossRef

20.

Booth AE, Seabra MC, Hume AN. Rab27a and melanosomes: a model to investigate the membrane targeting of Rabs. Biochem Soc Trans. 2012;40(6):1383–8.PubMedCrossRef

21.

Wang JS, Wang FB, Zhang QG, Shen ZZ, Shao ZM. Enhanced expression of Rab27A gene by breast cancer cells promoting invasiveness and the metastasis potential by secretion of insulin-like growth factor-II. Mol Cancer Res. 2008;6(3):372–82.PubMedCrossRef

22.

Nicotra G, Castino R, Follo C, Peracchio C, Valente G, Isidoro C. The dilemma: does tissue expression of cathepsin D reflect tumor malignancy? The question: does the assay truly mirror cathepsin D mis-function in the tumor? Cancer Biomark. 2010;7(1):47–64.PubMed

23.

Vetvicka V, Fusek M. Procathepsin D as a tumor marker, anti-cancer drug or screening agent. Anticancer Agents Med Chem. 2012;12(2):172–5.PubMedCrossRef

24.

Chlabicz M, Gacko M, Worowska A, Lapinski R. Cathepsin E (EC 3.4.23.34)—a review. Folia Histochem Cytobiol. 2011;49(4):547–57.PubMed

25.

He B, Yin B, Wang B, Xia Z, Chen C, Tang J. microRNAs in esophageal cancer (review). Mol Med Report. 2012;6(3):459–65.

26.

Link A, Kupcinskas J, Wex T, Malfertheiner P. Macro-role of microRNA in gastric cancer. Dig Dis. 2012;30(3):255–67.PubMedCrossRef

27.

Wang P, Chen L, Zhang J, Chen H, Fan J, Wang K, et al. Methylation-mediated silencing of the miR-124 genes facilitates pancreatic cancer progression and metastasis by targeting Rac1. Oncogene. 2013. doi:10.1038/onc.2012.598.

28.

Karsy M, Arslan E, Moy F. Current progress on understanding microRNAs in glioblastoma multiforme. Genes Cancer. 2012;3(1):3–15.PubMedCrossRef

29.

Ponomarev ED, Veremeyko T, Weiner HL. MicroRNAs are universal regulators of differentiation, activation, and polarization of microglia and macrophages in normal and diseased CNS. Glia. 2013;61(1):91–103.PubMedCrossRef

30.

Hirst TC, Vesterinen HM, Sena ES, Egan KJ, Macleod MR, Whittle IR. Systematic review and meta-analysis of temozolomide in animal models of glioma: was clinical efficacy predicted? Br J Cancer. 2013;108(1):64–71.PubMedCrossRef

31.

Moore G, Collins A, Brand C, Gold M, Lethborg C, Murphy M, et al. Palliative and supportive care needs of patients with high-grade glioma and their carers: A systematic review of qualitative literature. Patient Educ Couns. 2012. doi:10.1016/j.pec.2012.11.002.

32.

Sciume G, Santoni A, Bernardini G. Chemokines and glioma: invasion and more. J Neuroimmunol. 2010;224(1–2):8–12.PubMedCrossRef

33.

Sherman JH, Hoes K, Marcus J, Komotar RJ, Brennan CW, Gutin PH. Neurosurgery for brain tumors: update on recent technical advances. Curr Neurol Neurosci Rep. 2011;11(3):313–9.PubMedCrossRef

34.

Horgan CP, McCaffrey MW. Rab GTPases and microtubule motors. Biochem Soc Trans. 2011;39(5):1202–6.PubMedCrossRef

35.

Grosshans BL, Ortiz D, Novick P. Rabs and their effectors: achieving specificity in membrane traffic. Proc Natl Acad Sci U S A. 2006;103(32):11821–7.PubMedCrossRef

36.

Li Y, Hu J. Small GTPases and cilia. Protein Cell. 2011;2(1):13–25.PubMedCrossRef

37.

Hoffman NJ, Elmendorf JS. Signaling, cytoskeletal and membrane mechanisms regulating GLUT4 exocytosis. Trends Endocrinol Metab. 2011;22(3):110–6.PubMedCrossRef

38.

Itzen A, Goody RS. GTPases involved in vesicular trafficking: structures and mechanisms. Semin Cell Dev Biol. 2011;22(1):48–56.PubMedCrossRef

39.

Mitra S, Cheng KW, Mills GB. Rab GTPases implicated in inherited and acquired disorders. Semin Cell Dev Biol. 2011;22(1):57–68.PubMedCrossRef

40.

Agola J, Jim P, Ward H, Basuray S, Wandinger-Ness A. Rab GTPases as regulators of endocytosis, targets of disease and therapeutic opportunities. Clin Genet. 2011. doi:10.1111/j.399-0004.2011.01724.x.PubMed

41.

Bello-Morales R, Crespillo AJ, Fraile-Ramos A, Tabares E, Alcina A, Lopez-Guerrero JA. Role of the small GTPase Rab27a during Herpes simplex virus infection of oligodendrocytic cells. BMC Microbiol. 2012;12:265.PubMedCrossRef

42.

Bobrie A, Krumeich S, Reyal F, Recchi C, Moita LF, Seabra MC, et al. Rab27a supports exosome-dependent and -independent mechanisms that modify the tumor microenvironment and can promote tumor progression. Cancer Res. 2012;72(19):4920–30.PubMedCrossRef

43.

Peinado H, Aleckovic M, Lavotshkin S, Matei I, Costa-Silva B, Moreno-Bueno G, et al. Melanoma exosomes educate bone marrow progenitor cells toward a pro-metastatic phenotype through MET. Nat Med. 2012;18(6):883–91.PubMedCrossRef

44.

Hendrix A, Maynard D, Pauwels P, Braems G, Denys H, Van den Broecke R, et al. Effect of the secretory small GTPase Rab27B on breast cancer growth, invasion, and metastasis. J Natl Cancer Inst. 2010;102(12):866–80.PubMedCrossRef

45.

Dejeans N, Pluquet O, Lhomond S, Grise F, Bouchecareilh M, Juin A, et al. Autocrine control of glioma cells adhesion and migration through IRE1alpha-mediated cleavage of SPARC mRNA. J Cell Sci. 2012;125(Pt 18):4278–87.PubMedCrossRef

46.

Chen L, Chen XR, Zhang R, Li P, Liu Y, Yan K, et al. MicroRNA-107 inhibits glioma cell migration and invasion by modulating Notch2 expression. J Neurooncol. 2013. doi:10.1007/s11060-012-1037-7.

47.

Culine S, Honore N, Closson V, Droz JP, Extra JM, Marty M, et al. A small GTP-binding protein is frequently overexpressed in peripheral blood mononuclear cells from patients with solid tumours. Eur J Cancer. 1994;30A(5):670–4.PubMedCrossRef

48.

Pruitt FL, He Y, Franco OE, Jiang M, Cates JM, Hayward SW. Cathepsin D acts as an essential mediator to promote malignancy of benign prostatic epithelium. Prostate. 2013;73:476–88.PubMedCrossRef

49.

Laurent-Matha V, Huesgen PF, Masson O, Derocq D, Prebois C, Gary-Bobo M, et al. Proteolysis of cystatin C by cathepsin D in the breast cancer microenvironment. FASEB J. 2012;26(12):5172–81.PubMedCrossRef

50.

Kirana C, Shi H, Laing E, Hood K, Miller R, Bethwaite P, et al. Cathepsin D expression in colorectal cancer: from proteomic discovery through validation using western blotting, immunohistochemistry, and tissue microarrays. Int J Proteomics. 2012;2012:245819.PubMed

51.

Liu Y, Zhou Y, Zhu K. Inhibition of glioma cell lysosome exocytosis inhibits glioma invasion. PLoS One. 2012;7(9):e45910.PubMedCrossRef

Title: Effects of Rab27a on proliferation, invasion, and anti-apoptosis in human glioma cell
Authors: Xiuwei Wu
Anla Hu
Mingjun Zhang
Zhendong Chen
Publication date: 01-08-2013
Publisher: Springer Netherlands
Published in: Tumor Biology / Issue 4/2013
Print ISSN: 1010-4283
Electronic ISSN: 1423-0380
DOI: https://doi.org/10.1007/s13277-013-0756-5

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