All Title Author
Keywords Abstract

PLOS ONE  2013 

XB130, a New Adaptor Protein, Regulates Expression of Tumor Suppressive MicroRNAs in Cancer Cells

DOI: 10.1371/journal.pone.0059057

Full-Text   Cite this paper   Add to My Lib


XB130, a novel adaptor protein, promotes cell growth by controlling expression of many related genes. MicroRNAs (miRNAs), which are frequently mis-expressed in cancer cells, regulate expression of targeted genes. In this present study, we aimed to explore the oncogenic mechanism of XB130 through miRNAs regulation. We analyzed miRNA expression in XB130 short hairpin RNA (shRNA) stably transfected WRO thyroid cancer cells by a miRNA array assay, and 16 miRNAs were up-regulated and 22 miRNAs were down-regulated significantly in these cells, in comparison with non-transfected or negative control shRNA transfected cells. We chose three of the up-regulated miRNAs (miR-33a, miR-149 and miR-193a-3p) and validated them by real-time qRT-PCR. Ectopic overexpression of XB130 suppressed these 3 miRNAs in MRO cells, a cell line with very low expression of XB130. Furthermore, we transfected miR mimics of these 3 miRNAs into WRO cells. They negatively regulated expression of oncogenes (miR-33a: MYC, miR-149: FOSL1, miR-193a-3p: SLC7A5), by targeting their 3′ untranslated region, and reduced cell growth. Our results suggest that XB130 could promote growth of cancer cells by regulating expression of tumor suppressive miRNAs and their targeted genes.


[1]  Flynn DC (2001) Adaptor proteins. Oncogene 20: 6270–6272.
[2]  Yamanaka D, Akama T, Fukushima T, Nedachi T, Kawasaki C, et al. (2012) Phosphatidylinositol 3-kinase-binding protein, PI3KAP/XB130, is required for cAMP-induced amplification of IGF mitogenic activity in FRTL-5 thyroid cells. Mol Endocrinol 26: 1043–1055.
[3]  Shiozaki A, Liu M (2011) Roles of XB130, a novel adaptor protein, in cancer. J Clin Bioinforma 1: 10.
[4]  Csiszar A (2006) Structural and functional diversity of adaptor proteins involved in tyrosine kinase signalling. Bioessays 28: 465–479.
[5]  Dorfleutner A, Stehlik C, Zhang J, Gallick GE, Flynn DC (2007) AFAP-110 is required for actin stress fiber formation and cell adhesion in MDA-MB-231 breast cancer cells. J Cell Physiol 213: 740–749.
[6]  Zhang J, Park SI, Artime MC, Summy JM, Shah AN, et al. (2007) AFAP-110 is overexpressed in prostate cancer and contributes to tumorigenic growth by regulating focal contacts. J Clin Invest 117: 2962–2973.
[7]  Xu J, Bai XH, Lodyga M, Han B, Xiao H, et al. (2007) XB130, a novel adaptor protein for signal transduction. J Biol Chem 282: 16401–16412.
[8]  Lodyga M, De Falco V, Bai XH, Kapus A, Melillo RM, et al. (2009) XB130, a tissue-specific adaptor protein that couples the RET/PTC oncogenic kinase to PI 3-kinase pathway. Oncogene 28: 937–949.
[9]  Shiozaki A, Shen-Tu G, Bai X, Iitaka D, De Falco V, et al. (2012) XB130 mediates cancer cell proliferation and survival through multiple signaling events downstream of Akt. PLoS One 7: e43646.
[10]  Shiozaki A, Lodyga M, Bai XH, Nadesalingam J, Oyaizu T, et al. (2011) XB130, a novel adaptor protein, promotes thyroid tumor growth. Am J Pathol 178: 391–401.
[11]  Lodyga M, Bai XH, Kapus A, Liu M (2010) Adaptor protein XB130 is a Rac-controlled component of lamellipodia that regulates cell motility and invasion. J Cell Sci 123: 4156–4169.
[12]  Shiozaki A, Kosuga T, Ichikawa D, Komatsu S, Fujiwara H, et al. (2012) XB130 as an Independent Prognostic Factor in Human Esophageal Squamous Cell Carcinoma. Ann Surg Oncol.
[13]  Shi M, Huang W, Lin L, Zheng D, Zuo Q, et al. (2012) Silencing of XB130 is associated with both the prognosis and chemosensitivity of gastric cancer. PLoS One 7: e41660.
[14]  Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116: 281–297.
[15]  Stefani G, Slack FJ (2008) Small non-coding RNAs in animal development. Nat Rev Mol Cell Biol 9: 219–230.
[16]  Schmittgen TD (2008) Regulation of microRNA processing in development, differentiation and cancer. J Cell Mol Med 12: 1811–1819.
[17]  Rottiers V, Naar AM (2012) MicroRNAs in metabolism and metabolic disorders. Nat Rev Mol Cell Biol 13: 239–250.
[18]  Bueno MJ, Malumbres M (2011) MicroRNAs and the cell cycle. Biochim Biophys Acta 1812: 592–601.
[19]  He L, Thomson JM, Hemann MT, Hernando-Monge E, Mu D, et al. (2005) A microRNA polycistron as a potential human oncogene. Nature 435: 828–833.
[20]  He L, He X, Lim LP, de Stanchina E, Xuan Z, et al. (2007) A microRNA component of the p53 tumour suppressor network. Nature 447: 1130–1134.
[21]  Lu J, Getz G, Miska EA, Alvarez-Saavedra E, Lamb J, et al. (2005) MicroRNA expression profiles classify human cancers. Nature 435: 834–838.
[22]  Calin GA, Croce CM (2006) MicroRNA signatures in human cancers. Nat Rev Cancer 6: 857–866.
[23]  McKinsey EL, Parrish JK, Irwin AE, Niemeyer BF, Kern HB, et al. (2011) A novel oncogenic mechanism in Ewing sarcoma involving IGF pathway targeting by EWS/Fli1-regulated microRNAs. Oncogene 30: 4910–4920.
[24]  Chang S, Wang RH, Akagi K, Kim KA, Martin BK, et al. (2011) Tumor suppressor BRCA1 epigenetically controls oncogenic microRNA-155. Nat Med 17: 1275–1282.
[25]  Suzuki HI, Yamagata K, Sugimoto K, Iwamoto T, Kato S, et al. (2009) Modulation of microRNA processing by p53. Nature 460: 529–533.
[26]  Lee DY, Jeyapalan Z, Fang L, Yang J, Zhang Y, et al. (2010) Expression of versican 3′-untranslated region modulates endogenous microRNA functions. PLoS One 5: e13599.
[27]  Geraldo MV, Yamashita AS, Kimura ET (2012) MicroRNA miR-146b-5p regulates signal transduction of TGF-beta by repressing SMAD4 in thyroid cancer. Oncogene 31: 1910–1922.
[28]  Colamaio M, Borbone E, Russo L, Bianco M, Federico A, et al. (2011) miR-191 down-regulation plays a role in thyroid follicular tumors through CDK6 targeting. J Clin Endocrinol Metab 96: E1915–1924.
[29]  Braun J, Hoang-Vu C, Dralle H, Huttelmaier S (2010) Downregulation of microRNAs directs the EMT and invasive potential of anaplastic thyroid carcinomas. Oncogene 29: 4237–4244.
[30]  Liu W, Guo M, Ezzat S, Asa SL (2011) Vitamin D inhibits CEACAM1 to promote insulin/IGF-I receptor signaling without compromising anti-proliferative action. Lab Invest 91: 147–156.
[31]  Lodyga M, Bai XH, Mourgeon E, Han B, Keshavjee S, et al. (2002) Molecular cloning of actin filament-associated protein: a putative adaptor in stretch-induced Src activation. Am J Physiol Lung Cell Mol Physiol 283: L265–274.
[32]  Han B, Bai XH, Lodyga M, Xu J, Yang BB, et al. (2004) Conversion of mechanical force into biochemical signaling. J Biol Chem 279: 54793–54801.
[33]  Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25: 402–408.
[34]  Xiao H, Bai XH, Wang Y, Kim H, Mak AS, et al. (2013) MEK/ERK pathway mediates PKC activation-induced recruitment of PKCzeta and MMP-9 to podosomes. J Cell Physiol 228: 416–427.
[35]  Shiozaki A, Bai XH, Shen-Tu G, Moodley S, Takeshita H, et al. (2012) Claudin 1 mediates TNFalpha-induced gene expression and cell migration in human lung carcinoma cells. PLoS One 7: e38049.
[36]  Thomas M, Lange-Grunweller K, Weirauch U, Gutsch D, Aigner A, et al. (2012) The proto-oncogene Pim-1 is a target of miR-33a. Oncogene 31: 918–928.
[37]  Liu H, Brannon AR, Reddy AR, Alexe G, Seiler MW, et al. (2010) Identifying mRNA targets of microRNA dysregulated in cancer: with application to clear cell Renal Cell Carcinoma. BMC Syst Biol 4: 51.
[38]  Uhlmann S, Mannsperger H, Zhang JD, Horvat EA, Schmidt C, et al. (2012) Global microRNA level regulation of EGFR-driven cell-cycle protein network in breast cancer. Mol Syst Biol 8: 570.
[39]  Yu Z, Jian Z, Shen SH, Purisima E, Wang E (2007) Global analysis of microRNA target gene expression reveals that miRNA targets are lower expressed in mature mouse and Drosophila tissues than in the embryos. Nucleic Acids Res 35: 152–164.
[40]  Calin GA, Croce CM (2006) MicroRNAs and chromosomal abnormalities in cancer cells. Oncogene 25: 6202–6210.
[41]  Kim VN (2005) MicroRNA biogenesis: coordinated cropping and dicing. Nat Rev Mol Cell Biol 6: 376–385.
[42]  Schweppe RE, Klopper JP, Korch C, Pugazhenthi U, Benezra M, et al. (2008) Deoxyribonucleic acid profiling analysis of 40 human thyroid cancer cell lines reveals cross-contamination resulting in cell line redundancy and misidentification. J Clin Endocrinol Metab 93: 4331–4341.
[43]  Alitalo K, Schwab M, Lin CC, Varmus HE, Bishop JM (1983) Homogeneously staining chromosomal regions contain amplified copies of an abundantly expressed cellular oncogene (c-myc) in malignant neuroendocrine cells from a human colon carcinoma. Proc Natl Acad Sci U S A 80: 1707–1711.
[44]  Cerutti J, Trapasso F, Battaglia C, Zhang L, Martelli ML, et al. (1996) Block of c-myc expression by antisense oligonucleotides inhibits proliferation of human thyroid carcinoma cell lines. Clin Cancer Res 2: 119–126.
[45]  Yoshioka K, Deng T, Cavigelli M, Karin M (1995) Antitumor promotion by phenolic antioxidants: inhibition of AP-1 activity through induction of Fra expression. Proc Natl Acad Sci U S A 92: 4972–4976.
[46]  Young MR, Colburn NH (2006) Fra-1 a target for cancer prevention or intervention. Gene 379: 1–11.
[47]  Kim YH, Oh JH, Kim NH, Choi KM, Kim SJ, et al. (2001) Fra-1 expression in malignant and benign thyroid tumor. Korean J Intern Med 16: 93–97.
[48]  Kobayashi H, Ishii Y, Takayama T (2005) Expression of L-type amino acid transporter 1 (LAT1) in esophageal carcinoma. J Surg Oncol 90: 233–238.
[49]  Furuya M, Horiguchi J, Nakajima H, Kanai Y, Oyama T (2012) Correlation of L-type amino acid transporter 1 and CD98 expression with triple negative breast cancer prognosis. Cancer Sci 103: 382–389.
[50]  Miko E, Margitai Z, Czimmerer Z, Varkonyi I, Dezso B, et al. (2011) miR-126 inhibits proliferation of small cell lung cancer cells by targeting SLC7A5. FEBS Lett 585: 1191–1196.
[51]  Kim CH, Park KJ, Park JR, Kanai Y, Endou H, et al. (2006) The RNA interference of amino acid transporter LAT1 inhibits the growth of KB human oral cancer cells. Anticancer Res 26: 2943–2948.
[52]  Bader AG, Brown D, Stoudemire J, Lammers P (2011) Developing therapeutic microRNAs for cancer. Gene Ther 18: 1121–1126.


comments powered by Disqus