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  • br Conclusions br A new analytical method for the accurate

    2020-08-12


    4. Conclusions
    A new analytical method for the accurate determination of trans-ferrin in cell cultures has been developed based on the use of iodinated 3-Deazaneplanocin A and ICP-MS linked sandwich immunoassay. The fast and efficient iodination of the antibody by using Iodo beads with minimum degradation was corroborated by the analysis of the obtained product that revealed and stoichiometry of 27:1 iodine moles per mole of an-tibody. In addition, no significant losses of recognition capabilities of the antibody were observed in further immune subtraction experiment using affinity beads. The iodinated antibody was further implemented into a simple and efficient sandwich assay in combination with a sec-ondary biotinylated antibody and streptavidin coated magnetic micro-particles. This strategy permitted the quantitative determination of transferrin in breast cancer cell lines of different malignancy by indirect determination of iodine by ICP-MS. The obtained results showed a correlation between the cells with larger concentration of transferrin and also higher level of cytosolic Fe detected in previous studies with the same cell lines.
    Acknowledgements
    The authors would like to acknowledge the regional funding from the Government of Asturias through the Science, Technology and Innovation Plan (PCTI) co-financed by FEDER funds (Ref. FC-15-GRUPIN-14-010) and the funding from the Spanish Ministry of Economy, Industry and Competitiveness (MINECO) through the project CTQ2016–80069-C 2-1R. 
    References
    [17] J. Alonso García, D. Turiel Fernández, E. Añón Alvarez, E. Blanco González,
    F.J. Alonso-García et al.
    [26] L. Zhengru, L. Xiaoting, X. Guangyang, C. Beibei, H. Man, H. Bin, Application of inductively coupled plasma mass spectrometry in the quantitative analysis of bio-molecules with exogenous tags: a review, TrAC, Trends Anal. Chem. 93 (2017)
    Contents lists available at ScienceDirect
    Gene
    journal homepage: www.elsevier.com/locate/gene
    Research paper
    An integrated approach for mining precise RNA-based cervical cancer staging biomarkers 
    T
    Satarupa Banerjee, Devarajan Karunagaran
    Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, IIT Madras, Chennai 600036, India
    Keywords:
    Cervical cancer
    miRNA
    lncRNA
    Biomarker
    FIGO staging 
    Since international federation of gynecology and obstetrics (FIGO) staging is mainly based on clinical assess-ment, an integrated approach for mining RNA based biomarkers for understanding the molecular deregulation of signaling pathways and RNAs in cervical cancer was proposed in this study. Publicly available data were mined for identifying significant RNAs after patient staging. Significant miRNA families were identified from mRNA-miRNA and lncRNA-miRNA interaction network analyses followed by stage specific mRNA-miRNA-lncRNA as-sociation network generation. Integrated bioinformatic analyses of selected mRNAs and lncRNAs were per-formed. Results suggest that HBA1, HBA2, HBB, SLC2A1, CXCL10 (stage I), PKIA (stage III) and S100A7 (stage IV) were important. miRNA family enrichment of interacting miRNA partners of selected RNAs indicated the enrichment of let-7 family. Assembly of collagen fibrils and other multimeric structures_Homosapiens_R-HSA-2022090 in pathway analysis and progesterone_CTD_00006624 in DSigDB analysis were the most significant and SLC2A1, hsa-miR-188-3p, hsa-miR-378a-3p and hsa-miR-150-5p were selected as survival markers.
    1. Introduction
    Cervical cancer (CC) is the second most common cancer (Ifemelumma et al., 2019) and third cause of cancer related deaths (Shafabakhsh et al., 2019) in females worldwide with high incidence and mortality in the age group of 5 to 44 (Sreedevi et al., 2015). Most of the CC associated deaths (85%) occur in low and middle income countries due to lack of infrastructure, screening, vaccination and treatment (LaVigne et al., 2017; Cohen et al., 2019). International federation of gynecology and obstetrics (FIGO) staging of CC is based on visual clinical assessment of disease advancement by the physician in different anatomical locations of the tumor and suffers from inter-observer variability further complicated by the multistep nature of CC progression (Benedet et al., 2000). Since this is completely subjective, objective methods for staging of CC are needed for a thorough under-standing of the molecular deregulation of signaling pathways during CC progression. Previous studies have integrated genomic and molecular information with human papilloma virus (HPV) status (Anon., 2017), but stage specific analysis of transcriptome profiles is yet to be per-formed. When diagnosed at early stages, five-year survival in CC pa-tients is around 65% and it can be maximised by identification of specific early staging markers for predicting poor prognosis besides diagnosis. High risk type viral infections (such as HPV-16 and HPV-18)