title: Differentiation Induces Dramatic Changes in miRNA Profile, Where Loss of Dicer Diverts Differentiating SH-SY5Y Cells Toward Senescence.
abstract: MicroRNAs (miRNAs) are generated by endonuclease activity of Dicer, which also helps in loading of miRNAs to their target sequences. SH-SY5Y, a human neuroblastoma and a cellular model of neurodevelopment, consistently expresses genes related to neurodegenerative disorders at different biological levels (DNA, RNA, and proteins). Using SH-SY5Y cells, we have studied the role of Dicer and miRNAs in neuronal differentiation and explored involvement of P53, a master regulator of gene expression in differentiation-induced induction of miRNAs. Knocking down Dicer gene induced senescence in differentiating SH-SY5Y cells, which indicate the essential role of Dicer in brain development. Differentiation of SH-SY5Y cells by retinoic acid (RA) or RA + brain-derived neurotrophic factor (BDNF) induced dramatic changes in global miRNA expression. Fully differentiated SH-SY5Y cells (5-day RA followed by 3-day BDNF) significantly (p < 0.05 and atleast >3-fold change) upregulated and downregulated the expression of 77 and 17 miRNAs, respectively. Maximum increase was observed in the expression of miR-193-5p, miR-199a-5p, miR-192, miR-145, miR-28-5p, miR-29b, and miR-222 after RA exposure and miR-193-5p, miR-146a, miR-21, miR-199a-5p, miR-153, miR-29b, and miR-222 after RA + BDNF exposure in SH-SY5Y cells. Exploring the role of P53 in differentiating SH-SY5Y cells, we have observed that induction of miR-222, miR-192, and miR-145 is P53 dependent and expression of miR-193a-5p, miR-199a-5p, miR-146a, miR-21, miR-153, and miR-29b is P53 independent. In conclusion, decreased Dicer level enforces differentiating cells to senescence, and differentiating SH-SY5Y cells needs increased expression of P53 to cope up with changes in protein levels of mature neurons.
authors.affiliation: Developmental Toxicology Laboratory, Systems Toxicology and Health Risk Assessment Group, CSIR-Indian Institute of Toxicology Research (IITR), Vishvigyan Bhawan, 31 Mahatma Gandhi Marg, Lucknow, UP, 226001, India.|Academy of Scientific and Innovative Research (AcSIR), IITR Campus, Lucknow, India.
authors.forename: Abhishek
authors.initials: A
authors.lastname: Jauhari
authors.affiliation: Developmental Toxicology Laboratory, Systems Toxicology and Health Risk Assessment Group, CSIR-Indian Institute of Toxicology Research (IITR), Vishvigyan Bhawan, 31 Mahatma Gandhi Marg, Lucknow, UP, 226001, India.|Department of Biochemistry, School of Dental Sciences, Babu Banarasi Das University, Lucknow, India.
authors.forename: Tanisha
authors.initials: T
authors.lastname: Singh
authors.affiliation: Developmental Toxicology Laboratory, Systems Toxicology and Health Risk Assessment Group, CSIR-Indian Institute of Toxicology Research (IITR), Vishvigyan Bhawan, 31 Mahatma Gandhi Marg, Lucknow, UP, 226001, India.
authors.forename: Ankita
authors.initials: A
authors.lastname: Pandey
authors.affiliation: Developmental Toxicology Laboratory, Systems Toxicology and Health Risk Assessment Group, CSIR-Indian Institute of Toxicology Research (IITR), Vishvigyan Bhawan, 31 Mahatma Gandhi Marg, Lucknow, UP, 226001, India.|Department of Biochemistry, School of Dental Sciences, Babu Banarasi Das University, Lucknow, India.
authors.forename: Parul
authors.initials: P
authors.lastname: Singh
authors.affiliation: Developmental Toxicology Laboratory, Systems Toxicology and Health Risk Assessment Group, CSIR-Indian Institute of Toxicology Research (IITR), Vishvigyan Bhawan, 31 Mahatma Gandhi Marg, Lucknow, UP, 226001, India.|Department of Biochemistry, School of Dental Sciences, Babu Banarasi Das University, Lucknow, India.
authors.forename: Nishant
authors.initials: N
authors.lastname: Singh
authors.affiliation: Developmental Toxicology Laboratory, Systems Toxicology and Health Risk Assessment Group, CSIR-Indian Institute of Toxicology Research (IITR), Vishvigyan Bhawan, 31 Mahatma Gandhi Marg, Lucknow, UP, 226001, India.|Academy of Scientific and Innovative Research (AcSIR), IITR Campus, Lucknow, India.
authors.forename: Ankur Kumar
authors.initials: AK
authors.lastname: Srivastava
authors.affiliation: Developmental Toxicology Laboratory, Systems Toxicology and Health Risk Assessment Group, CSIR-Indian Institute of Toxicology Research (IITR), Vishvigyan Bhawan, 31 Mahatma Gandhi Marg, Lucknow, UP, 226001, India.
authors.forename: Aditya Bhushan
authors.initials: AB
authors.lastname: Pant
authors.affiliation: Developmental Toxicology Laboratory, Systems Toxicology and Health Risk Assessment Group, CSIR-Indian Institute of Toxicology Research (IITR), Vishvigyan Bhawan, 31 Mahatma Gandhi Marg, Lucknow, UP, 226001, India.
authors.forename: Devendra
authors.initials: D
authors.lastname: Parmar
authors.affiliation: Developmental Toxicology Laboratory, Systems Toxicology and Health Risk Assessment Group, CSIR-Indian Institute of Toxicology Research (IITR), Vishvigyan Bhawan, 31 Mahatma Gandhi Marg, Lucknow, UP, 226001, India. sanjay@iitr.res.in.
authors.forename: Sanjay
authors.initials: S
authors.lastname: Yadav
mesh_terms: D019208:Brain-Derived Neurotrophic Factor / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D016922:Cellular Senescence; D053487:DEAD-box RNA Helicases / Q000378:metabolism*; D015536:Down-Regulation / Q000187:drug effects; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D035683:MicroRNAs / Q000378:metabolism*; D009447:Neuroblastoma / Q000378:metabolism; D055495:Neurogenesis / Q000187:drug effects; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D043244:Ribonuclease III / Q000378:metabolism*; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D035683:MicroRNAs; D014212:Tretinoin; C579455:BDNF protein, human; C408087:DICER1 protein, human; D043244:Ribonuclease III; D053487:DEAD-box RNA Helicases
keywords: Dicer; MicroRNAs; Neuronal differentiation; P53; SH-SY5Y; Senescence