Open AccessFunctional analysis of microRNAs in human hepatocellular cancer stem cells
Author(s) -
Meng Fanyin,
Glaser Shan S.,
Francis Heather,
DeMorrow Sharon,
Han Yuyan,
Passarini Jenna D.,
Stokes Allison,
Cleary John P.,
Liu Xiuping,
Venter Julie,
Kumar Preetham,
Priester Sally,
Hubble Levi,
Staloch Dustin,
Sharma Jay,
Liu ChangGong,
Alpini Gianfranco
Publication year - 2012
Publication title -
journal of cellular and molecular medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.44
H-Index - 130
eISSN - 1582-4934
pISSN - 1582-1838
DOI - 10.1111/j.1582-4934.2011.01282.x
Subject(s) - microrna , biology , gene silencing , stem cell , cancer research , gene expression profiling , gene expression , regulation of gene expression , microarray analysis techniques , cancer stem cell , microbiology and biotechnology , gene , genetics
MicroRNAs are endogenous small non‐coding RNAs that regulate gene expression and cancer development. A rare population of hepatocellular cancer stem cells (HSCs) holds the extensive proliferative and self‐renewal potential necessary to form a liver tumour. We postulated that specific transcriptional factors might regulate the expression of microRNAs and subsequently modulate the expression of gene products involved in phenotypic characteristics of HSCs. We evaluated the expression of microRNA in human HSCs by microarray profiling, and defined the target genes and functional effects of two groups of microRNA regulated by IL‐6 and transcriptional factor Twist. A subset of highly chemoresistant and invasive HSCs was screened with aberrant expressions of cytokine IL‐6 and Twist. We demonstrated that conserved let‐7 and miR‐181 family members were up‐regulated in HSCs by global microarray‐based microRNA profiling followed by validation with real‐time polymerase chain reaction. Importantly, inhibition of let‐7 increases the chemosensitivity of HSCs to sorafenib and doxorubicin whereas silencing of miR‐181 led to a reduction in HSCs motility and invasion. Knocking down IL‐6 and Twist in HSCs significantly reduced let‐7 and miR‐181 expression and subsequently inhibited chemoresistance and cell invasion. We showed that let‐7 directly targets SOCS‐1 and caspase‐3, whereas miR‐181 directly targets RASSF1A, TIMP3 as well as nemo‐like kinase (NLK). In conclusion, alterations of IL‐6‐ and Twist‐regulated microRNA expression in HSCs play a part in tumour spreading and responsiveness to chemotherapy. Our results define a novel regulatory mechanism of let‐7/miR‐181s suggesting that let‐7 and miR‐181 may be molecular targets for eradication of hepatocellular malignancies.