miR-589-5p inhibits MAP3K8 and suppresses CD90+cancer stem cells in hepatocellular carcinoma.
Cancer stem cells (CSCs) are important in the tumorigenesis and progression of hepatocellular carcinoma (HCC). MicroRNAs (miRNAs) play crucial roles regulating CD133+and EpCAM+CSCs in HCC, although it is unclear whether miRNAs regulate CD90+CSCs in HCC.



The miRNA profiles of CD90+and CD90-HCC cells were analyzed using a miRNA microarray and quantitative real-time PCR (qRT-PCR). CSC characteristics were examined by qRT-PCR and Western blot of pluripotency-associated genes, clone and sphere formation assay, transwell migration assay, and nude mice tumorigenicity assay. miR-589-5p mimic transfection was used to overexpress miR-589-5p in vitro. The CD90 and miR-589-5p expressions of HCC samples were detected by immunohistochemistry and qRT-PCR, respectively.



miR-589-5p and miR-33b-5p were down-regulated in CD90+cells. Overexpression of miR-589-5p suppressed CD90+CSC characteristics such as Oct4, Sox2 and Nanog expression, a high likelihood of forming cell spheres, high invasiveness and high tumorigenicity. Luciferase reporter assays demonstrated that miR-589-5p directly binds to the 3'-untranslated region of mitogen-activated protein kinase kinase kinase 8 (MAP3K8) mRNA, and exogenous miR-589-5p down-regulated MAP3K8 expression. In addition, siRNA inhibition of MAP3K8 also suppressed CD90+CSC characteristics, even in the absence of miR-589-5p overexpression. In HCC tissues, miR-589-5p expression was inversely correlated with CD90 expression, and high CD90 expression and low miR-589-5p expression were positively correlated with vascular invasion and recurrence and significantly decreased disease-free and overall survival by clinical analysis.



In HCC, miR-589-5p down-regulates the stemness characteristics of CD90+CSCs in part by silencing MAP3K8. CD90 and miR-589-5p expression predict HCC outcomes and might be novel molecular targets for HCC treatment.
Institute of Hepatobiliary Surgery, Southwest Hospital, Third Military Medical University, Chongqing, 400037, China.
Xi
X
Zhang
Institute of Hepatobiliary Surgery, Southwest Hospital, Third Military Medical University, Chongqing, 400037, China.
Peng
P
Jiang
Institute of Hepatobiliary Surgery, Southwest Hospital, Third Military Medical University, Chongqing, 400037, China.
Ling
L
Shuai
Institute of Hepatobiliary Surgery, Southwest Hospital, Third Military Medical University, Chongqing, 400037, China.
Kai
K
Chen
Institute of Hepatobiliary Surgery, Southwest Hospital, Third Military Medical University, Chongqing, 400037, China.
Zhonghu
Z
Li
Institute of Hepatobiliary Surgery, Southwest Hospital, Third Military Medical University, Chongqing, 400037, China.
Yujun
Y
Zhang
Institute of Hepatobiliary Surgery, Southwest Hospital, Third Military Medical University, Chongqing, 400037, China.
Yan
Y
Jiang
Institute of Hepatobiliary Surgery, Southwest Hospital, Third Military Medical University, Chongqing, 400037, China. lixw1966@163.com.
Xiaowu
X
Li
D000328:Adult; D000368:Aged; D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D045744:Cell Line, Tumor; D015536:Down-Regulation; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D020930:MAP Kinase Kinase Kinases / Q000235:genetics*; D008297:Male; D051379:Mice; D008819:Mice, Nude; D035683:MicroRNAs / Q000235:genetics*; D008875:Middle Aged; D009368:Neoplasm Transplantation; D014411:Neoplastic Stem Cells / Q000378:metabolism / Q000473:pathology*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D018800:Thy-1 Antigens / Q000378:metabolism*
C550416:MIRN33a microRNA, human; C586366:MIRN589 microRNA, human; D035683:MicroRNAs; D011518:Proto-Oncogene Proteins; D018800:Thy-1 Antigens; D020930:MAP Kinase Kinase Kinases; C069937:MAP3K8 protein, human
CD90; Cellular heterogeneity; MAP3K8; Prognosis; Tumorigenicity; microRNA