Breast cancer cell line MDA-MB-231 miRNA profile expression after BIK interference: BIK involvement in autophagy
Por:
Ruiz Esparza-Garrido, Ruth, Eugenia Torres-Marquez, Maria, Viedma-Rodriguez, Rubi, Claudia Velazquez-Wong, Ana, Salamanca-Gomez, Fabio, Rosas-Vargas, Haydee, Angel Velazquez-Flores, Miguel
Publicada:
1 may 2016
Categoría:
Cancer Research
Resumen:
B-cell lymphoma 2 (BCL2)-interacting killer (apoptosis inducing) (BIK)
has been proposed as a tumor suppressor in diverse types of cancers.
However, BIK's overexpression in breast cancer (BC) and in non-small
lung cancer cells (NSCLCs), associated with a poor prognosis, suggests
its participation in tumor progression. In this study, we evaluated the
global expression pattern of microRNAs (miRNAs), messenger RNA (mRNA)
expression changes in autophagy, and autophagic flux after BIK
interference. BIK gene expression was silenced by small interfering RNA
(siRNA) in BC cell MDA-MB-231, and BIK interference efficiency was
tested by real-time PCR and by Western blotting. BIK expression levels
decreased by 75 +/- 18 % in the presence of 600 nM siRNA, resulting in
the abolishment of BIK expression by 94 +/- 30 %. BIK interference
resulted in the overexpression of 17 miRNAs that, according to the
DIANA-miRPath v3.0 database, are mainly implied in the control of cell
signaling, gene expression, and autophagy. The autophagy array revealed
downregulation of transcripts which participate in autophagy, and their
interactome revealed a complex network, where hepatocyte growth
factor-regulated tyrosine kinase substrate (HGS), alpha-synuclein
(SNCA), unc-51-like autophagy activating kinase 1/2 (ULK1/2), and
mitogen-activated protein kinase 3 (MAPK3) were shown to be signaling
hubs. LC3-II expression-an autophagy marker-was increased by 169 +/- 25
% after BIK interference, which indicates the involvement of BIK in
autophagy. Altogether, our results indicate-for the first time-that BIK
controls the expression of miRNAs, as well as the autophagic flux in
MDA-MB-231 cells.
Filiaciones:
Mexican Inst Social Secur IMSS, Natl Med Ctr Century 21, Childrens Hosp, Funct Genom Lab,Unit Human Genet Res, Mexico City 06720, DF, Mexico
Univ Nacl Autonoma Mexico, Sch Med, Dept Biochem, Mexico City 04510, DF, Mexico
Univ Nacl Autonoma Mexico UNAM Mexico, FES Iztacala, Unidad Morfol & Func Celular, Lab Biol Desarrollo, Tlalnepantla 54090, Estado De Mexic, Mexico
Inst Mexicano Seguro Social, Ctr Med Nacl Siglo 21, Coordinac Invest Salud, Mexico City, DF, Mexico
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