Expression and function of c1orf132 long-noncoding rna in breast cancer cell lines and tissues

Shafaroudi, A. M ; Sharif University of Technology | 2021

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  1. Type of Document: Article
  2. DOI: 10.3390/ijms22136768
  3. Publisher: MDPI , 2021
  4. Abstract:
  5. miR-29b2 and miR-29c play a suppressive role in breast cancer progression. C1orf132 (also named MIR29B2CHG) is the host gene for generating both microRNAs. However, the region also expresses longer transcripts with unknown functions. We employed bioinformatics and experimental approaches to decipher C1orf132 expression and function in breast cancer tissues. We also used the CRISPR/Cas9 technique to excise a predicted C1orf132 distal promoter and followed the behavior of the edited cells by real-time PCR, flow cytometry, migration assay, and RNA-seq techniques. We observed that C1orf132 long transcript is significantly downregulated in triple-negative breast cancer. We also identified a promoter for the longer transcripts of C1orf132 whose functionality was demonstrated by transfecting MCF7 cells with a C1orf132 promoter-GFP construct. Knocking-out the promoter by means of CRISPR/Cas9 revealed no alterations in the expression of the neighboring genes CD46 and CD34, while the expression of miR-29c was reduced by half. Furthermore, the promoter knockout elevated the migration ability of the edited cells. RNA sequencing revealed many up-and downregulated genes involved in various cellular pathways, including epithelial to mesenchymal transition and mammary gland development pathways. Altogether, we are reporting here the existence of an additional/distal promoter with an enhancer effect on miR-29 generation and an inhibitory effect on cell migration. © 2021 by the authors. Licensee MDPI, Basel, Switzerland
  6. Keywords:
  7. CD34 antigen ; Long noncoding RNA c1orf132 ; Membrane cofactor protein ; MicroRNA ; MicroRNA 29 ; microRNA 29b2 ; MicroRNA 29c ; Unclassified drug ; Long untranslated RNA ; MicroRNA ; MIRN29B1 microRNA, human ; MIRN29C microRNA, human ; Bioinformatics ; Breast cancer ; Breast development ; Breast tissue ; Cancer tissue ; Cell migration assay ; Controlled study ; CRISPR-CAS9 system ; Down regulation ; Enhancer region ; Epithelial mesenchymal transition ; Flow cytometry ; Gene knockout ; Human ; Human cell ; Human tissue ; Mammary gland ; MCF-7 cell line ; Promoter region ; Protein expression ; protein function ; Real time polymerase chain reaction ; RNA sequencing ; Triple negative breast cancer ; Upregulation ; Animal ; Cell nucleus ; CRISPR Cas system ; Gene expression regulation ; Gene knockdown ; Genetics ; Metabolism ; Mouse ; Tumor cell line ; Animals ; Cell Line, Tumor ; Cell Nucleus ; CRISPR-Cas Systems ; Down-Regulation ; Epithelial-Mesenchymal Transition ; Female ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Gene Knockdown Techniques ; Humans ; MCF-7 Cells ; Mice ; MicroRNAs ; Promoter Regions, Genetic ; RNA, Long Noncoding ; Triple Negative Breast Neoplasms
  8. Source: International Journal of Molecular Sciences ; Volume 22, Issue 13 , 2021 ; 16616596 (ISSN)
  9. URL: https://www.mdpi.com/1422-0067/22/13/6768