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Cited by 92MicroRNA in disease regulationCell death mechanisms and regulationCancer-related Molecular Pathways

A novel microRNA mmu-miR-466h affects apoptosis regulation in mammalian cells

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Druz, Aliaksandr, Chu, Chia, Majors, Brian, Santuary, Rodell, Betenbaugh, Michael, Shiloach, Joseph·2011·Biotechnology and Bioengineering
DOI 10.1002/bit.23092PMC PMC3099433
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Abstract

This study determined the changes in microRNA (miRs) expression in mammalian Chinese hamster ovary (CHO) cells undergoing apoptosis induced by exposing the cells to nutrient‐depleted media. The apoptosis onset was confirmed by reduced cell viability and Caspase‐3/7 activation. Microarray comparison of known mouse and rat miRs in CHO cells exposed to fresh or depleted media revealed up‐regulation of the mouse miR‐297‐669 cluster in CHO cells subjected to depleted media. The mmu‐miR‐466h was chosen for further analysis as the member of this cluster with the highest overexpression and its up‐regulation in depleted media was confirmed with qRT‐PCR. Since miRs suppress mRNA translation, we hypothesized that up‐regulated mmu‐miR‐466h inhibits anti‐apoptotic genes and induces apoptosis. A combination of bioinformatics and experimental tools was used to predict and verify mmu‐miR‐466h anti‐apoptotic targets. 8708 predicted targets were obtained from miRecords database and narrowed to 38 anti‐apoptotic genes with DAVID NCBI annotation tool. Several genes were selected from this anti‐apoptotic subset based on nucleotide pairing complimentarity between the mmu‐miR‐466h seed region and 3′ UTR of the target mRNAs. The qRT‐PCR analysis revealed reduced mRNA levels of bcl2l2, dad1, birc6, stat5a, and smo genes in CHO cells exposed to depleted media. The inhibition of the mmu‐miR‐466h increased the expression levels of those genes and resulted in increased cell viability and decreased Caspase‐3/7 activation. The up‐regulation of mmu‐miR‐466h in response to nutrients depletion causes the inhibition of several anti‐apoptotic genes in unison. This suggests the pro‐apoptotic role of mmu‐miR‐466h and its capability to modulate the apoptotic pathway in mammalian cells. Biotechnol. Bioeng. 2011; 108:1651–1661. © 2011 Wiley Periodicals, Inc.

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  • Inhibition of the mmu-miR-466 h microRNA was shown to increase apoptosis resistance of these cells.

    “Also, as discussed earlier, inhibition of the mmu-miR-466 h microRNA was shown to increase apoptosis resistance of these cells (Druz et al., 2011;Druz et al., 2013).”

    From Therapeutic glycoprotein production in mammalian cells (2017) · cites this paper as Druz et al., 2011

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Research works printed but not linked: 75

  1. Aharinejad S Andrukhova O Lucas T Zuckermann A Wieselthaler G Wolner E Grimm M 2008 Programmed cell death in idiopathic dilated cardiomyopathy is mediated by suppression of the apoptosis inhibitor Apollon Ann Thorac Surg 86 1 109 14 discussion 114 18573408
  2. Akao Y Nakagawa Y Naoe T 2006 let-7 microRNA functions as a potential growth suppressor in human colon cancer cells Biol Pharm Bull 29 5 903 6 16651716
  3. Arden N Betenbaugh MJ 2004 Life and death in mammalian cell culture: strategies for apoptosis inhibition Trends Biotechnol 22 4 174 80 15038922
  4. Arden N Majors BS Ahn SH Oyler G Betenbaugh MJ 2007 Inhibiting the apoptosis pathway using MDM2 in mammalian cell cultures Biotechnol Bioeng 97 3 601 14 17149774
  5. Babak T Zhang W Morris Q Blencowe BJ Hughes TR 2004 Probing microRNAs with microarrays: tissue specificity and functional inference RNA 10 11 1813 9 15496526 PMC1370668
  6. Barisic K Petrik J Rumora L 2003 Biochemistry of apoptotic cell death Acta Pharm 53 3 151 64 14769239
  7. Bialik S Cryns VL Drincic A Miyata S Wollowick AL Srinivasan A Kitsis RN 1999 The mitochondrial apoptotic pathway is activated by serum and glucose deprivation in cardiac myocytes Circ Res 85 5 403 14 10473670
  8. Blackburn RV Spitz DR Liu X Galoforo SS Sim JE Ridnour LA Chen JC Davis BH Corry PM Lee YJ 1999 Metabolic oxidative stress activates signal transduction and gene expression during glucose deprivation in human tumor cells Free Radic Biol Med 26 3–4 419 30 9895234
  9. Cai X Hagedorn CH Cullen BR 2004 Human microRNAs are processed from capped, polyadenylated transcripts that can also function as mRNAs RNA 10 12 1957 66 15525708 PMC1370684
  10. Carmeliet P Dor Y Herbert JM Fukumura D Brusselmans K Dewerchin M Neeman M Bono F Abramovitch R Maxwell P 1998 Role of HIF-1alpha in hypoxia-mediated apoptosis, cell proliferation and tumour angiogenesis Nature 394 6692 485 90 9697772
  11. Chen XL Cao LQ She MR Wang Q Huang XH Fu XH 2008 Gli-1 siRNA induced apoptosis in Huh7 cells World J Gastroenterol 14 4 582 9 18203291 PMC2681150
  12. Cheng AM Byrom MW Shelton J Ford LP 2005 Antisense inhibition of human miRNAs and indications for an involvement of miRNA in cell growth and apoptosis Nucleic Acids Res 33 4 1290 7 15741182 PMC552951
  13. Chiang GG Sisk WP 2005 Bcl-x(L) mediates increased production of humanized monoclonal antibodies in Chinese hamster ovary cells Biotechnol Bioeng 91 7 779 92 15986489
  14. Chigancas V Batista LF Brumatti G Amarante-Mendes GP Yasui A Menck CF 2002 Photorepair of RNA polymerase arrest and apoptosis after ultraviolet irradiation in normal and XPB deficient rodent cells Cell Death Differ 9 10 1099 107 12232798
  15. Crawford M Batte K Yu L Wu X Nuovo GJ Marsh CB Otterson GA Nana-Sinkam SP 2009 MicroRNA 133B targets pro-survival molecules MCL-1 and BCL2L2 in lung cancer Biochem Biophys Res Commun 388 3 483 9 19654003 PMC2824514
  16. Enright AJ John B Gaul U Tuschl T Sander C Marks DS 2003 MicroRNA targets in Drosophila Genome Biol 5 1 R1 14709173 PMC395733
  17. Faraoni I Antonetti FR Cardone J Bonmassar E 2009 miR-155 gene: a typical multifunctional microRNA Biochim Biophys Acta 1792 6 497 505 19268705
  18. Figueroa B Jr Ailor E Osborne D Hardwick JM Reff M Betenbaugh MJ 2007 Enhanced cell culture performance using inducible anti-apoptotic genes E1B-19K and Aven in the production of a monoclonal antibody with Chinese hamster ovary cells Biotechnol Bioeng 97 4 877 92 17099908
  19. Figueroa B Jr Chen S Oyler GA Hardwick JM Betenbaugh MJ 2004 Aven and Bcl-xL enhance protection against apoptosis for mammalian cells exposed to various culture conditions Biotechnol Bioeng 85 6 589 600 14966800
  20. Fribley A Zhang K Kaufman RJ 2009 Regulation of apoptosis by the unfolded protein response Methods Mol Biol 559 191 204 19609758 PMC3684430
  21. Fujinaka Y Takane K Yamashita H Vasavada RC 2007 Lactogens promote beta cell survival through JAK2/STAT5 activation and Bcl-XL upregulation J Biol Chem 282 42 30707 17 17728251
  22. Gammell P 2007 MicroRNAs: recently discovered key regulators of proliferation and apoptosis in animal cells: Identification of miRNAs regulating growth and survival Cytotechnology 53 1–3 55 63 19003190 PMC2267611
  23. Gammell P Barron N Kumar N Clynes M 2007 Initial identification of low temperature and culture stage induction of miRNA expression in suspension CHO-K1 cells J Biotechnol 130 3 213 8 17570552
  24. Garofalo M Quintavalle C Di Leva G Zanca C Romano G Taccioli C Liu CG Croce CM Condorelli G 2008 MicroRNA signatures of TRAIL resistance in human non-small cell lung cancer Oncogene 27 27 3845 55 18246122
  25. Hayashita Y Osada H Tatematsu Y Yamada H Yanagisawa K Tomida S Yatabe Y Kawahara K Sekido Y Takahashi T 2005 A polycistronic microRNA cluster, miR-17-92, is overexpressed in human lung cancers and enhances cell proliferation Cancer Res 65 21 9628 32 16266980
  26. Hutvagner G Zamore PD 2002 A microRNA in a multiple-turnover RNAi enzyme complex Science 297 5589 2056 60 12154197
  27. Janumyan YM Sansam CG Chattopadhyay A Cheng N Soucie EL Penn LZ Andrews D Knudson CM Yang E 2003 Bcl-xL/Bcl-2 coordinately regulates apoptosis, cell cycle arrest and cell cycle entry EMBO J 22 20 5459 70 14532118 PMC213787
  28. Jovanovic M Hengartner MO 2006 miRNAs and apoptosis: RNAs to die for Oncogene 25 46 6176 87 17028597
  29. Kantardjieff A Nissom PM Chuah SH Yusufi F Jacob NM Mulukutla BC Yap M Hu WS 2009 Developing genomic platforms for Chinese hamster ovary cells Biotechnol Adv 27 6 1028 35 19470403
  30. Kim R 2005 Unknotting the roles of Bcl-2 and Bcl-xL in cell death Biochem Biophys Res Commun 333 2 336 43 15922292
  31. Kroemer G Reed JC 2000 Mitochondrial control of cell death Nat Med 6 5 513 9 10802706
  32. Kuhn DE Martin MM Feldman DS Terry AV Jr Nuovo GJ Elton TS 2008 Experimental validation of miRNA targets Methods 44 1 47 54 18158132 PMC2237914
  33. Kumar MS Lu J Mercer KL Golub TR Jacks T 2007 Impaired microRNA processing enhances cellular transformation and tumorigenesis Nat Genet 39 5 673 7 17401365
  34. Lanceta J Prough RA Liang R Wang E 2010 MicroRNA group disorganization in aging Exp Gerontol 45 4 269 78 20034554
  35. Lee Y Ahn C Han J Choi H Kim J Yim J Lee J Provost P Radmark O Kim S 2003 The nuclear RNase III Drosha initiates microRNA processing Nature 425 6956 415 9 14508493
  36. Lee YJ Galoforo SS Berns CM Tong WP Kim HR Corry PM 1997 Glucose deprivation-induced cytotoxicity in drug resistant human breast carcinoma MCF-7/ADR cells: role of c-myc and bcl-2 in apoptotic cell death J Cell Sci 110 Pt 5 681 6 9092950
  37. Li J Smyth P Cahill S Denning K Flavin R Aherne S Pirotta M Guenther SM O’Leary JJ Sheils O 2008 Improved RNA quality and TaqMan Pre-amplification method (PreAmp) to enhance expression analysis from formalin fixed paraffin embedded (FFPE) materials BMC Biotechnol 8 10 18254955 PMC2259333
  38. Li W Ruan K 2009 MicroRNA detection by microarray Anal Bioanal Chem 394 4 1117 24 19132354
  39. Liang R Bates DJ Wang E 2009 Epigenetic Control of MicroRNA Expression and Aging Curr Genomics 10 3 184 93 19881911 PMC2705851
  40. Lim SF Chuan KH Liu S Loh SO Chung BY Ong CC Song Z 2006 RNAi suppression of Bax and Bak enhances viability in fed-batch cultures of CHO cells Metab Eng 8 6 509 22 16860584
  41. Lim Y Wong NS Lee YY Ku SC Wong DC Yap MG 2010 Engineering mammalian cells in bioprocessing - current achievements and future perspectives Biotechnol Appl Biochem 55 4 175 89 20392202
  42. Lynam-Lennon N Maher SG Reynolds JV 2009 The roles of microRNA in cancer and apoptosis Biol Rev Camb Philos Soc 84 1 55 71 19046400
  43. Maes OC An J Sarojini H Wang E 2008 Murine microRNAs implicated in liver functions and aging process Mech Ageing Dev 129 9 534 41 18561983
  44. Majors BS Betenbaugh MJ Chiang GG 2007 Links between metabolism and apoptosis in mammalian cells: applications for anti-apoptosis engineering Metab Eng 9 4 317 26 17611135
  45. Makishima T Yoshimi M Komiyama S Hara N Nishimoto T 2000 A subunit of the mammalian oligosaccharyltransferase, DAD1, interacts with Mcl-1, one of the bcl-2 protein family J Biochem 128 3 399 405 10965038
  46. Mastrangelo AJ Betenbaugh MJ 1998 Overcoming apoptosis: new methods for improving protein-expression systems Trends Biotechnol 16 2 88 95 9487737
  47. Mendell JT 2005 MicroRNAs: critical regulators of development, cellular physiology and malignancy Cell Cycle 4 9 1179 84 16096373
  48. Mercille S Massie B 1994 Induction of apoptosis in nutrient-deprived cultures of hybridoma and myeloma cells Biotechnol Bioeng 44 9 1140 54 18623032
  49. Mercille S Massie B 1999 Apoptosis-resistant E1B-19K-expressing NS/0 myeloma cells exhibit increased viability and chimeric antibody productivity under perfusion culture conditions Biotechnol Bioeng 63 5 529 43 10397809
  50. Moley KH Mueckler MM 2000 Glucose transport and apoptosis Apoptosis 5 2 99 105 11232248
  51. Mott JL Kobayashi S Bronk SF Gores GJ 2007 mir-29 regulates Mcl-1 protein expression and apoptosis Oncogene 26 42 6133 40 17404574 PMC2432524
  52. Muller D Katinger H Grillari J 2008 MicroRNAs as targets for engineering of CHO cell factories Trends Biotechnol 26 7 359 65 18471912
  53. Nissom PM 2007 Specific detection of residual CHO host cell DNA by real-time PCR Biologicals 35 3 211 5 17071102
  54. Nivitchanyong T Martinez A Ishaque A Murphy JE Konstantinov K Betenbaugh MJ Thrift J 2007 Anti-apoptotic genes Aven and E1B-19K enhance performance of BHK cells engineered to express recombinant factor VIII in batch and low perfusion cell culture Biotechnol Bioeng 98 4 825 41 17514750
  55. Rehmsmeier M Steffen P Hochsmann M Giegerich R 2004 Fast and effective prediction of microRNA/target duplexes RNA 10 10 1507 17 15383676 PMC1370637
  56. Reynolds JE Yang T Qian L Jenkinson JD Zhou P Eastman A Craig RW 1994 Mcl-1, a member of the Bcl-2 family, delays apoptosis induced by c-Myc overexpression in Chinese hamster ovary cells Cancer Res 54 24 6348 52 7987827
  57. Roldo C Missiaglia E Hagan JP Falconi M Capelli P Bersani S Calin GA Volinia S Liu CG Scarpa A 2006 MicroRNA expression abnormalities in pancreatic endocrine and acinar tumors are associated with distinctive pathologic features and clinical behavior J Clin Oncol 24 29 4677 84 16966691
  58. Rossi D Gaidano G 2003 Messengers of cell death: apoptotic signaling in health and disease Haematologica 88 2 212 8 12604411
  59. Sanjay A Fu J Kreibich G 1998 DAD1 is required for the function and the structural integrity of the oligosaccharyltransferase complex J Biol Chem 273 40 26094 9 9748289
  60. Schwarz DS Hutvagner G Du T Xu Z Aronin N Zamore PD 2003 Asymmetry in the assembly of the RNAi enzyme complex Cell 115 2 199 208 14567917
  61. Tang Y Zheng J Sun Y Wu Z Liu Z Huang G 2009 MicroRNA-1 regulates cardiomyocyte apoptosis by targeting Bcl-2 Int Heart J 50 3 377 87 19506341
  62. Thorburn A 2004 Death receptor-induced cell killing Cell Signal 16 2 139 44 14636884
  63. Trummer E Ernst W Hesse F Schriebl K Lattenmayer C Kunert R Vorauer-Uhl K Katinger H Muller D 2008 Transcriptional profiling of phenotypically different Epo-Fc expressing CHO clones by cross-species microarray analysis Biotechnol J 3 7 924 37 18481264
  64. Vecchione A Croce CM 2009 Apoptomirs: small molecules have gained the license to kill Endocr Relat Cancer 17 1 F37 50 19815577
  65. Werner RG Noe W Kopp K Schluter M 1998 Appropriate mammalian expression systems for biopharmaceuticals Arzneimittelforschung 48 8 870 80 9748718
  66. Wong DC Wong KT Nissom PM Heng CK Yap MG 2006 Targeting early apoptotic genes in batch and fed-batch CHO cell cultures Biotechnol Bioeng 95 3 350 61 16894638
  67. Xiao F Zuo Z Cai G Kang S Gao X Li T 2009 miRecords: an integrated resource for microRNA-target interactions Nucleic Acids Res 37 Database issue D105 10 18996891 PMC2686554
  68. Xu J Chen S Chen H Xiao Q Hsu CY Michael D Bao J 2009a STAT5 mediates antiapoptotic effects of methylprednisolone on oligodendrocytes J Neurosci 29 7 2022 6 19228956 PMC2662324
  69. Xu XF Guo CY Liu J Yang WJ Xia YJ Xu L Yu YC Wang XP 2009b Gli1 maintains cell survival by up-regulating IGFBP6 and Bcl-2 through promoter regions in parallel manner in pancreatic cancer cells J Carcinog 8 13 19736394 PMC2746911
  70. Yin KJ Deng Z Huang H Hamblin M Xie C Zhang J Chen YE 2010 miR-497 regulates neuronal death in mouse brain after transient focal cerebral ischemia Neurobiol Dis 38 1 17 26 20053374 PMC2837803
  71. Yoshimi M Sekiguchi T Hara N Nishimoto T 2000 Inhibition of N-linked glycosylation causes apoptosis in hamster BHK21 cells Biochem Biophys Res Commun 276 3 965 9 11027576
  72. Yue D Liu H Huang Y 2009 Survey of Computational Algorithms for MicroRNA Target Prediction Curr Genomics 10 7 478 92 20436875 PMC2808675
  73. Yue J Tigyi G 2010 Conservation of miR-15a/16-1 and miR-15b/16-2 clusters Mamm Genome 21 1–2 88 94 20013340 PMC2820079
  74. Zanghi JA Fussenegger M Bailey JE 1999 Serum protects protein-free competent Chinese hamster ovary cells against apoptosis induced by nutrient deprivation in batch culture Biotechnol Bioeng 64 1 108 19 10397845
  75. Zhu Q Hong A Sheng N Zhang X Matejko A Jun KY Srivannavit O Gulari E Gao X Zhou X 2007 microParaflo biochip for nucleic acid and protein analysis Methods Mol Biol 382 287 312 18220239

Cited by

Sorted by citations

Authors
Druz, A., Chu, C., Majors, B., Santuary, R., Betenbaugh, M., & Shiloach, J.
Affiliations
  • National Institutes of Health (US)
  • Johns Hopkins University (US)
Corresponding author
  • Betenbaugh, Michael
  • Shiloach, Joseph
ORCID
Venue
Biotechnology and Bioengineering
Publisher
Wiley
Published
2011-03-11
Volume
108
Issue
7
Pages
1651-1661
ISSN
  • 0006-3592 (ISSN-L)
  • 1097-0290
DOI
10.1002/bit.23092
PMC
PMC3099433
OpenAlex
W2128560728
Cited by
92
References
1
Citations by year
2026: 2 · 2025: 3 · 2023: 2 · 2022: 11 · 2021: 7 · 2020: 9 · 2019: 5 · 2018: 5 · 2017: 3 · 2016: 6 · 2015: 8 · 2014: 10 · 2013: 8 · 2012: 12
FWCI
3.37
Type
journal article
Language
en
Open access
Licence
onlinelibrary.wiley.com/termsAndConditions#vor (vor)
Locations
  • Biotechnology and Bioengineering (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • National Institutes of Health
  • National Institute of Diabetes and Digestive and Kidney Diseases (Z01DK070010)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › MicroRNA in disease regulation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cell death mechanisms and regulation
  • Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
Keywords
microRNA, apoptosis, nutrient deprivation, CHO cells, anti-apoptotic genes, BCL2L2, DAD1, BIRC6, STAT5A, SMO, caspase-3/7 activation, gene expression profiling, miRNA target prediction
MeSH terms
Animals, Cell Survival, Cricetulus, Culture Media, Food, Gene Expression Regulation, Cricetinae, CHO Cells, Apoptosis, Reverse Transcriptase Polymerase Chain Reaction, Gene Expression Profiling, MicroRNAs, Mice, Rats, Caspase 3, Caspase 7

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