In vitro selections of mammaglobin A and mammaglobin B aptamers for the recognition of circulating breast tumor cells

Mammaglobin B (MGB2) and mammaglobin A (MGB1) are proteins expressed in metastatic breast cancers. The early detection of circulating tumor cells (CTCs) in breast cancer patients is crucial to decrease mortality rate. Herein, novel aptamers were successfully selected and characterized against MGB2 and MGB1 proteins using a hybrid SELEX approach. The potential use of the selected aptamers in breast CTC detection was studied using spiked breast cancer cells in whole blood lysate. The results obtained from this study showed that the selected aptamers (MAMB1 and MAMA2) bind to their target breast cancer cell lines with high affinity (low nanomolar Kd values) and specificity. They also bind to their free recombinant target proteins and show minimal non-specific binding to normal and other cancer cell lines. Additionally, they were able to distinguish a low number of breast cancer cells spiked in whole blood lysate containing normal blood cells. The results obtained in this study indicate the great potential for the use of aptamers to detect MGB1 and MGB2 protein biomarkers, expressed on the surface of breast CTCs.

[1]  F. Graham,et al.  Characteristics of a human cell line transformed by DNA from human adenovirus type 5. , 1977, The Journal of general virology.

[2]  J. Szostak,et al.  In vitro selection of RNA molecules that bind specific ligands , 1990, Nature.

[3]  M. Watson,et al.  Mammaglobin, a mammary-specific member of the uteroglobin gene family, is overexpressed in human breast cancer. , 1996, Cancer research.

[4]  B. Glasgow,et al.  Lipophilin, a novel heterodimeric protein of human tears , 1998, FEBS letters.

[5]  M. Watson,et al.  Identification of mammaglobin B, a novel member of the uteroglobin gene family. , 1998, Genomics.

[6]  B. Glasgow,et al.  Lipophilins: human peptides homologous to rat prostatein. , 1999, Biochemical and biophysical research communications.

[7]  M. Watson,et al.  Mammaglobin expression in primary, metastatic, and occult breast cancer. , 1999, Cancer research.

[8]  O. Zach,et al.  Detection of circulating mammary carcinoma cells in the peripheral blood of breast cancer patients via a nested reverse transcriptase polymerase chain reaction assay for mammaglobin mRNA. , 1999, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[9]  A. Jemal,et al.  Breast Cancer Statistics , 2013 .

[10]  H. M. Beier,et al.  Uteroglobin/Clara Cell 10‐kDa Family of Proteins: Nomenclature Committee Report , 2000, Annals of the New York Academy of Sciences.

[11]  M Monden,et al.  Selection of mRNA markers for detection of lymph node micrometastases in breast cancer patients. , 2000, Oncology reports.

[12]  P. Friedman,et al.  Mammaglobin is found in breast tissue as a complex with BU101. , 2001, Biochemistry.

[13]  Ying-Fon Chang,et al.  Tenascin-C Aptamers Are Generated Using Tumor Cells and Purified Protein* , 2001, The Journal of Biological Chemistry.

[14]  K. Thompson,et al.  Pharmacokinetics and Biodistribution of Novel Aptamer Compositions , 2004, Pharmaceutical Research.

[15]  RT-PCR for mammaglobin genes, MGB1 and MGB2, identifies breast cancer micrometastases in sentinel lymph nodes. , 2004, American journal of clinical pathology.

[16]  D. Carter,et al.  Mammaglobin: a candidate diagnostic marker for breast cancer. , 2004, Clinical biochemistry.

[17]  J. Massagué,et al.  Epithelial-Mesenchymal Transitions Twist in Development and Metastasis , 2004, Cell.

[18]  C. Rogers IARC handbooks of cancer prevention, volume 7: breast cancer screening , 2004 .

[19]  A. Ballestrero,et al.  Effect of different cytokines on mammaglobin and maspin gene expression in normal leukocytes: possible relevance to the assays for the detection of micrometastatic breast cancer , 2005, British Journal of Cancer.

[20]  David W. Russell,et al.  Purification of nucleic acids by extraction with phenol:chloroform. , 2006, CSH protocols.

[21]  杨朝勇 Aptamers evolved from live cells as effective molecular probes for cancer study , 2006 .

[22]  Bertrand Tavitian,et al.  Comparison of different strategies to select aptamers against a transmembrane protein target. , 2006, Oligonucleotides.

[23]  S. Calza,et al.  Mammaglobin B expression in human endometrial cancer , 2007, International Journal of Gynecologic Cancer.

[24]  R. Cress,et al.  Descriptive analysis of estrogen receptor (ER)‐negative, progesterone receptor (PR)‐negative, and HER2‐negative invasive breast cancer, the so‐called triple‐negative phenotype , 2007, Cancer.

[25]  Tanja Fehm,et al.  Stem cell and epithelial-mesenchymal transition markers are frequently overexpressed in circulating tumor cells of metastatic breast cancer patients , 2009, Breast Cancer Research.

[26]  Hui Chen,et al.  Molecular recognition of acute myeloid leukemia using aptamers , 2008, Leukemia.

[27]  Ryan Walsh,et al.  Retention of function in the DNA homolog of the RNA dopamine aptamer. , 2009, Biochemical and biophysical research communications.

[28]  上官棣华 Development of DNA aptamers using Cell-SELEX , 2010 .

[29]  M. P. Ceballos,et al.  Detection of mammaglogin A in blood from breast cancer patients, before and after treatment, using a one-tube nested PCR protocol. Association with the absence of tumor estrogen receptors. , 2011, Clinical biochemistry.

[30]  Fazlul H. Sarkar,et al.  Cancer Stem Cells and Epithelial-to-Mesenchymal Transition (EMT)-Phenotypic Cells: Are They Cousins or Twins? , 2011, Cancers.

[31]  C. Dai,et al.  Mechanism of the Mesenchymal–Epithelial Transition and Its Relationship with Metastatic Tumor Formation , 2011, Molecular Cancer Research.

[32]  Mathew W. Wright,et al.  Update of the human secretoglobin (SCGB) gene superfamily and an example of 'evolutionary bloom' of androgen-binding protein genes within the mouse Scgb gene superfamily , 2011, Human Genomics.

[33]  S. Ohuchi,et al.  Cell-SELEX Technology , 2012, BioResearch open access.

[34]  M. Roizen,et al.  Hallmarks of Cancer: The Next Generation , 2012 .

[35]  B. Sullenger,et al.  Further characterization of the target of a potential aptamer biomarker for pancreatic cancer: cyclophilin B and its posttranslational modifications. , 2013, Nucleic acid therapeutics.

[36]  A. Redig,et al.  Breast cancer as a systemic disease: a view of metastasis , 2013, Journal of internal medicine.

[37]  N. Zhao,et al.  Immunotherapy of CD30-expressing lymphoma using a highly stable ssDNA aptamer. , 2013, Biomaterials.

[38]  J. Byun,et al.  Nucleic Acid Aptamers: New Methods for Selection, Stabilization, and Application in Biomedical Science , 2013, Biomolecules & therapeutics.

[39]  W. Duan,et al.  Selection of DNA aptamers against epithelial cell adhesion molecule for cancer cell imaging and circulating tumor cell capture. , 2013, Analytical chemistry.

[40]  E. Siegel,et al.  Mammaglobin B (SCGB2A1) is a novel tumour antigen highly differentially expressed in all major histological types of ovarian cancer: implications for ovarian cancer immunotherapy , 2013, British Journal of Cancer.

[41]  C. Mathers,et al.  GLOBOCAN 2012 v1.0, Cancer Incidence and Mortality Worldwide: IARC CancerBase No. 11 [Internet]. Lyon, France: International Agency for Research on Cancer , 2013 .

[42]  S. Tope,et al.  Aptamers as therapeutics , 2013 .

[43]  Etsuko Miyamoto-Sato,et al.  Towards Personalized Medicine Mediated by in Vitro Virus-Based Interactome Approaches , 2014, International journal of molecular sciences.

[44]  A. Tolcher,et al.  Phase 1 trial of the oral AKT inhibitor MK-2206 plus carboplatin/paclitaxel, docetaxel, or erlotinib in patients with advanced solid tumors , 2014, Journal of Hematology & Oncology.

[45]  Huang Jun,et al.  Descriptive analysis of estrogen receptor (ER)-negative, progesterone receptor (PR)-negative, and HER2-negative invasive breast cancer, the so-called triple-negative phenotype , 2014 .

[46]  Young-Pil Kim,et al.  Detection and Characterization of Cancer Cells and Pathogenic Bacteria Using Aptamer-Based Nano-Conjugates , 2014, Sensors.

[47]  Robert B Livingston,et al.  Circulating tumor cells and response to chemotherapy in metastatic breast cancer: SWOG S0500. , 2014, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[48]  Y. Zu,et al.  Oligonucleotide Aptamers: New Tools for Targeted Cancer Therapy , 2014, Molecular therapy. Nucleic acids.

[49]  Min Zhang,et al.  Developing aptamer probes for acute myelogenous leukemia detection and surface protein biomarker discovery , 2014, Journal of Hematology & Oncology.

[50]  S. Gopinath,et al.  Aptamers as a replacement for antibodies in enzyme-linked immunosorbent assay. , 2015, Biosensors & bioelectronics.

[51]  C. Takahashi,et al.  Detection of human mammaglobin A mRNA in peripheral blood of breast cancer patients before treatment and association with metastasis. , 2015, Journal of the Egyptian National Cancer Institute.

[52]  Ailiang Chen,et al.  Replacing antibodies with aptamers in lateral flow immunoassay. , 2015, Biosensors & bioelectronics.

[53]  Amphun Chaiboonchoe,et al.  The in vitro selection world. , 2016, Methods.

[54]  Eslie Dennis,et al.  The assessment of HER2 status in breast cancer: the past, the present, and the future , 2016, Pathology international.