Expression and Function of C1orf132 Long-Noncoding RNA in Breast Cancer Cell Lines and Tissues

被引:11
|
作者
Shafaroudi, Afsaneh Malekzadeh [1 ]
Sharifi-Zarchi, Ali [2 ]
Rahmani, Saeid [2 ]
Nafissi, Nahid [3 ]
Mowla, Seyed Javad [4 ]
Lauria, Andrea [5 ,6 ]
Oliviero, Salvatore [5 ,6 ]
Matin, Maryam M. [1 ,7 ]
机构
[1] Ferdowsi Univ Mashhad, Fac Sci, Dept Biol, Mashhad 9177948974, Razavi Khorasan, Iran
[2] Sharif Univ Technol, Dept Comp Engn, Tehran 1115511365, Iran
[3] Iran Univ Med Sci, Sch Med, Surg Dept, Tehran 1449614535, Iran
[4] Tarbiat Modares Univ, Fac Biol Sci, Dept Mol Genet, Tehran 14115154, Iran
[5] Univ Turin, Dept Life Sci & Syst Biol, I-10126 Turin, Italy
[6] Italian Inst Genom Med, I-10060 Candiolo, Italy
[7] Ferdowsi Univ Mashhad, Inst Biotechnol, Novel Diagnost & Therapeut Res Grp, Mashhad 9177948974, Razavi Khorasan, Iran
关键词
lncRNA; breast cancer; TNBC; EMT; miR-29; C1orf132; WNT GENE-EXPRESSION; MICRORNA-29; FAMILY; DNA;
D O I
10.3390/ijms22136768
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
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页数:14
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