PMID- 25983556
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20150518
LR  - 20220330
IS  - 1177-9322 (Print)
IS  - 1177-9322 (Electronic)
IS  - 1177-9322 (Linking)
VI  - 9
DP  - 2015
TI  - Computational and in vitro Investigation of miRNA-Gene Regulations in 
      Retinoblastoma Pathogenesis: miRNA Mimics Strategy.
PG  - 89-101
LID - 10.4137/BBI.S21742 [doi]
AB  - PURPOSE: Retinoblastoma (RB), a primary pediatric intraocular tumor, arises from 
      primitive retinal layers. Several novel molecular strategies are being developed 
      for the clinical management of RB. miRNAs are known to regulate cancer-relevant 
      biological processes. Here, the role of selected miRNAs, namely, miR-532-5p and 
      miR-486-3p, has been analyzed for potential therapeutic targeting in RB. METHODS: 
      A comprehensive bioinformatic analysis was performed to predict the 
      posttranscriptional regulators (miRNAs) of the select panel of genes [Group 1: 
      oncogenes (HMGA2, MYCN, SYK, FASN); Group 2: cancer stem cell markers (TACSTD, 
      ABCG2, CD133, CD44, CD24) and Group 3: cell cycle regulatory proteins (p53, 
      MDM2)] using Microcosm, DIANALAB, miRBase v 18, and REFSEQ database, and RNA 
      hybrid. The expressions of five miRNAs, namely, miR-146b-5p, miR-532-5p, 
      miR-142-5p, miR-328, and miR-486-3p, were analyzed by qRT-PCR on primary RB tumor 
      samples (n = 30; including 17 invasive RB tumors and 13 noninvasive RB tumors). 
      Detailed complementary alignment between 5' seed sequence of differentially 
      expressed miRNAs and the sequence of target genes was determined. Based on 
      minimum energy level and piCTAR scores, the gene targets were selected. 
      Functional roles of these miRNA clusters were studied by using mimics in cultured 
      RB (Y79, Weri Rb-1) cells in vitro. The gene targets (SYK and FASN) of the 
      studied miRNAs were confirmed by qRT-PCR and western blot analysis. Cell 
      proliferation and apoptotic studies were performed. RESULTS: Nearly 1948 miRNAs 
      were identified in the in silico analysis, From this list, only 9 upregulated 
      miRNAs (miR-146b-5p, miR-305, miR-663b, miR-299, miR-532-5p, miR-892b, miR-501, 
      miR-142-5p, and miR-513b) and 10 downregulated miRNAs (miR-1254, miR-328, 
      miR-133a, miR-1287, miR-1299, miR-375, miR-486-3p, miR-720, miR-98, and miR-122*) 
      were found to be common with the RB serum miRNA profile. Downregulation of five 
      miRNAs (miR-146b-5p, miR-532-5p, miR-142-5p, miR-328, and miR-486-3p) was 
      confirmed experimentally. Predicted common oncogene targets (SYK and FASN) of 
      miR-486-3p and miR-532-5p were evaluated for their mRNA and protein expression in 
      these miRNA mimic-treated RB cells. Experimental overexpression of these miRNAs 
      mediated apoptotic cell death without significantly altering the cell cycle in RB 
      cells. CONCLUSION: Key miRNAs in RB pathogenesis were identified by an in silico 
      approach. Downregulation of miR-486-3p and miR-532-5p in primary retinoblastoma 
      tissues implicates their role in tumorigenesis. Prognostic and therapeutic 
      potential of these miRNA was established by the miRNA mimic strategy.
FAU - Venkatesan, Nalini
AU  - Venkatesan N
AD  - Larsen & Toubro Department of Ocular Pathology, Vision Research Foundation, 
      Sankara Nethralaya, Chennai, India. ; Department of Biological Sciences, Birla 
      Institute of Technology and Science (BITS), Pilani, Rajasthan, India.
FAU - Deepa, Perinkulam Ravi
AU  - Deepa PR
AD  - Department of Biological Sciences, Birla Institute of Technology and Science 
      (BITS), Pilani, Rajasthan, India.
FAU - Khetan, Vikas
AU  - Khetan V
AD  - Department of Vitreoretina and Oncology, Medical Research Foundation, Sankara 
      Nethralaya, Chennai, India.
FAU - Krishnakumar, Subramanian
AU  - Krishnakumar S
AD  - Larsen & Toubro Department of Ocular Pathology, Vision Research Foundation, 
      Sankara Nethralaya, Chennai, India.
LA  - eng
PT  - Journal Article
DEP - 20150512
PL  - United States
TA  - Bioinform Biol Insights
JT  - Bioinformatics and biology insights
JID - 101467187
PMC - PMC4429751
OTO - NOTNLM
OT  - bio-informatics analysis
OT  - miRNA-mRNA
OT  - mimics
OT  - retinoblastoma
EDAT- 2015/05/20 06:00
MHDA- 2015/05/20 06:01
PMCR- 2015/05/12
CRDT- 2015/05/19 06:00
PHST- 2014/11/11 00:00 [received]
PHST- 2015/01/13 00:00 [revised]
PHST- 2015/01/17 00:00 [accepted]
PHST- 2015/05/19 06:00 [entrez]
PHST- 2015/05/20 06:00 [pubmed]
PHST- 2015/05/20 06:01 [medline]
PHST- 2015/05/12 00:00 [pmc-release]
AID - bbi-9-2015-089 [pii]
AID - 10.4137/BBI.S21742 [doi]
PST - epublish
SO  - Bioinform Biol Insights. 2015 May 12;9:89-101. doi: 10.4137/BBI.S21742. 
      eCollection 2015.