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Functional Role of MicroRNA‐200 Family in Human Gall Bladder Cancer Stem Cells
Author(s) -
Meng Fanyin,
Francis Heather,
Han Yuyan,
Glaser Shan,
Alpini Gianfranco
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.45.7
Subject(s) - microrna , epigenetics , biology , bladder cancer , meg3 , cancer research , cancer stem cell , methylation , stem cell , cancer , gene silencing , dna methylation , microbiology and biotechnology , gene expression , genetics , downregulation and upregulation , gene , long non coding rna
BACKGROUND & AIMS MicroRNAs (miRNAs) are highly conserved 19‐25 nucleotide noncoding small RNAs which are primarily involved in gene silencing. Our aim was to characterize the functional roles of epigenetic regulated microRNAs in human gall bladder cancer stem cells. METHODS miRNA expression in human gall bladder cancer tissues, human gall bladder cancer stem cells (GB‐CSCs), Mz‐ChA‐1 gall bladder cancer cells and H69 non‐malignant cholangiocytes was assessed using a hybridization based microarray and was further evaluated by Taqman real‐time PCR analysis. Promoter methylation, matrix metalloproteinase (MMP) 1, 2, 3 and 9 mRNA expression was quantitated by real‐time PCR analysis. RESULTS We identified miR‐200 family including miR‐200a, miR‐200b and miR‐200c that are differentially expressed in doxorubicin‐enriched cell fractions in a mouse xenograft tumor model of gall bladder cancer. Expression of miR‐200 family microRNAs was substantially enhanced concomitant with GBCs differentiation and loss of self‐renewal. Methylation‐specific PCR detected that miR‐200b and miR‐200c promoter was hypermethylated in GB‐CSCs, GBCs as well as Mz‐ChA‐1 cells. Cell migration and invasion were significantly decreased by miR‐200 precursor introduction in GB‐CSCs and Mz‐ChA‐1 cells. Bioinformatics and dual‐luciferase reporter assays identified ZEB1 and ZEB2 as the direct targets of miR‐200 family. CONCLUSION Our results define a novel epigenetic regulatory mechanism of specific microRNA group and suggesting that miR‐200 cluster may be the molecular targets for eradication of human gall bladder cancer.