Changes in the Expression of miR-381 and miR-495 Are Inversely Associated with the Expression of the MDR1 Gene and Development of Multi-Drug Resistance

Multidrug resistance (MDR) frequently develops in cancer patients exposed to chemotherapeutic agents and is usually brought about by over-expression of P-glycoprotein (P-gp) which acts as a drug efflux pump to reduce the intracellular concentration of the drug(s). Thus, inhibiting P-gp expression might assist in overcoming MDR in cancer chemotherapy. MiRNAome profiling using next-generation sequencing identified differentially expressed microRNAs (miRs) between parental K562 cells and MDR K562 cells (K562/ADM) induced by adriamycin treatment. Two miRs, miR-381 and miR-495, that were strongly down-regulated in K562/ADM cells, are validated to target the 3’-UTR of the MDR1 gene. These miRs are located within a miR cluster located at chromosome region 14q32.31, and all miRs in this cluster appear to be down-regulated in K562/ADM cells. Functional analysis indicated that restoring expression of miR-381 or miR-495 in K562/ADM cells was correlated with reduced expression of the MDR1 gene and its protein product, P-gp, and increased drug uptake by the cells. Thus, we have demonstrated that changing the levels of certain miR species modulates the MDR phenotype in leukemia cells, and propose further exploration of the use of miR-based therapies to overcome MDR.

[1]  K. Livak,et al.  Real-time quantification of microRNAs by stem–loop RT–PCR , 2005, Nucleic acids research.

[2]  Xiuping Liu,et al.  Role of MicroRNA miR-27a and miR-451 in the regulation of MDR1/P-glycoprotein expression in human cancer cells. , 2008, Biochemical pharmacology.

[3]  D. Greenblatt,et al.  Rapid Assessment of P-Glycoprotein Inhibition and Induction in Vitro , 2003, Pharmaceutical Research.

[4]  X. Bai,et al.  GCS induces multidrug resistance by regulating apoptosis-related genes in K562/AO2 cell line , 2010, Cancer Chemotherapy and Pharmacology.

[5]  R. Hellens,et al.  Quantitative stem-loop RT-PCR for detection of microRNAs. , 2011, Methods in molecular biology.

[6]  A. Seluanov,et al.  DNA repair by homologous recombination, but not by nonhomologous end joining, is elevated in breast cancer cells. , 2009, Neoplasia.

[7]  F. Russel,et al.  The Role of ATP Binding Cassette Transporters in Tissue Defense and Organ Regeneration , 2009, Journal of Pharmacology and Experimental Therapeutics.

[8]  T. Ando,et al.  Decitabine (5-Aza-2′-deoxycytidine) decreased DNA methylation and expression of MDR-1 gene in K562/ADM cells , 2000, Leukemia.

[9]  I. Pastan,et al.  Genetic analysis of the multidrug transporter. , 1995, Annual review of genetics.

[10]  François Chevenet,et al.  OligoHeatMap (OHM): an online tool to estimate and display hybridizations of oligonucleotides onto DNA sequences , 2008, Nucleic Acids Res..

[11]  C. Croce,et al.  Targeting microRNAs in cancer: rationale, strategies and challenges , 2010, Nature Reviews Drug Discovery.

[12]  Jared P. Stephens,et al.  Comparative analysis of different laser systems to study cellular responses to DNA damage in mammalian cells , 2009, Nucleic acids research.

[13]  Weixiong Zhang,et al.  Characterization and Identification of MicroRNA Core Promoters in Four Model Species , 2007, PLoS Comput. Biol..

[14]  Peter A. Jones,et al.  Epigenetics in human disease and prospects for epigenetic therapy , 2004, Nature.

[15]  Ima Paydar,et al.  Genomic Organization of Zebrafish microRNAs , 2008, BMC Genomics.

[16]  Reversal of ABC drug transporter-mediated multidrug resistance in cancer cells: evaluation of current strategies. , 2008, Current molecular pharmacology.

[17]  Anna Seelig,et al.  Quantification and characterization of P-glycoprotein-substrate interactions. , 2006, Biochemistry.

[18]  J. Hughes,et al.  Eukaryotic ribosomal RNA: the recent excitement in the nucleotide modification problem , 1997, Chromosoma.

[19]  Ana M. Aransay,et al.  miRanalyzer: a microRNA detection and analysis tool for next-generation sequencing experiments , 2009, Nucleic Acids Res..

[20]  A. Seluanov,et al.  Comparison of nonhomologous end joining and homologous recombination in human cells. , 2008, DNA repair.

[21]  U. Brinkmann,et al.  Functional polymorphisms of the human multidrug-resistance gene: multiple sequence variations and correlation of one allele with P-glycoprotein expression and activity in vivo. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[22]  M. Gottesman,et al.  Targeting multidrug resistance in cancer , 2006, Nature Reviews Drug Discovery.

[23]  T. Fojo,et al.  The role of ABC transporters in clinical practice. , 2003, The oncologist.

[24]  Stijn van Dongen,et al.  miRBase: tools for microRNA genomics , 2007, Nucleic Acids Res..

[25]  C. Croce,et al.  Human microRNA genes are frequently located at fragile sites and genomic regions involved in cancers. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[26]  Fei Li,et al.  Primary transcripts and expressions of mammal intergenic microRNAs detected by mapping ESTs to their flanking sequences , 2006, Mammalian Genome.

[27]  S. Korsmeyer,et al.  Cloning the chromosomal breakpoint of t(14;18) human lymphomas: clustering around Jh on chromosome 14 and near a transcriptional unit on 18 , 1985, Cell.

[28]  Jinhui Wu,et al.  Overcoming multidrug-resistance in vitro and in vivo using the novel P-glycoprotein inhibitor 1416 , 2012, Bioscience reports.

[29]  Andrey Golubov,et al.  Alterations of microRNAs and their targets are associated with acquired resistance of MCF‐7 breast cancer cells to cisplatin , 2010, International journal of cancer.

[30]  M. Gottesman.  Mechanisms of cancer drug resistance. , 2002, Annual review of medicine.

[31]  M. F. Shannon,et al.  A double-negative feedback loop between ZEB1-SIP1 and the microRNA-200 family regulates epithelial-mesenchymal transition. , 2008, Cancer research.

[32]  K. Ueda,et al.  A T3587G germ-line mutation of the MDR1 gene encodes a nonfunctional P-glycoprotein , 2006, Molecular Cancer Therapeutics.

[33]  Martin S. Taylor,et al.  The transcriptional network that controls growth arrest and differentiation in a human myeloid leukemia cell line , 2009, Nature Genetics.

[34]  D. Bartel.  MicroRNAs Genomics, Biogenesis, Mechanism, and Function , 2004, Cell.

[35]  Anna Seelig,et al.  Quantification and characterization of P-glycoprotein-substrate interactions. , 2006, Biochemistry.

[36]  J. G. Patton,et al.  Genomic organization of microRNAs , 2009, Journal of cellular physiology.

[37]  W. Huber,et al.  which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. MAnorm: a robust model for quantitative comparison of ChIP-Seq data sets , 2011 .

[38]  Anton J. Enright,et al.  Genomic analysis of human microRNA transcripts , 2007, Proceedings of the National Academy of Sciences.

[39]  N. Kamada,et al.  Chromosome abnormalities in adult T-cell leukemia/lymphoma: a karyotype review committee report. , 1992, Cancer research.

[40]  C. Croce.  Causes and consequences of microRNA dysregulation in cancer , 2009, Nature Reviews Genetics.

[41]  Chemosensitizers.  MODULATION OF P-GLYCOPROTEIN MEDIATED MULTIDRUG RESISTANCE ( MDR ) IN CANCER USING , 2010 .

[42]  M. Seto,et al.  Genome‐wide microRNA expression profiling in renal cell carcinoma: significant down‐regulation of miR‐141 and miR‐200c , 2008, The Journal of pathology.

[43]  J. Petriz,et al.  Flow cytometric analysis of P-glycoprotein function using rhodamine 123 , 1997, Leukemia.

[44]  A. Komamine,et al.  RNAi and Plant Gene Function Analysis , 2011, Methods in Molecular Biology.

[45]  Anton J. Enright,et al.  Human MicroRNA Targets , 2004, PLoS biology.

[46]  T. Tsuruo,et al.  Overcoming of vincristine resistance in P388 leukemia in vivo and in vitro through enhanced cytotoxicity of vincristine and vinblastine by verapamil. , 1981, Cancer research.

[47]  B. Cullen.  Transcription and processing of human microRNA precursors. , 2004, Molecular cell.

[48]  Olga Kovalchuk,et al.  Involvement of microRNA-451 in resistance of the MCF-7 breast cancer cells to chemotherapeutic drug doxorubicin , 2008, Molecular Cancer Therapeutics.

[49]  R. Berger,et al.  HOX11L2/TLX3 is transcriptionally activated through T-cell regulatory elements downstream of BCL11B as a result of the t(5;14)(q35;q32). , 2006, Blood.

[50]  Marwan Shinawi,et al.  Prader-Willi phenotype caused by paternal deficiency for the HBII-85 C/D box small nucleolar RNA cluster , 2008, Nature Genetics.

[51]  B. Meyers,et al.  Construction of small RNA cDNA libraries for deep sequencing. , 2007, Methods.

[52]  Martina Paulsen,et al.  Identification of tandemly-repeated C/D snoRNA genes at the imprinted human 14q32 domain reminiscent of those at the Prader-Willi/Angelman syndrome region. , 2002, Human molecular genetics.