[Changes of microRNAs profiling in mesothelial cells exposed to multi-walled carbon nanotubes].
Objective:To explore the cytotoxicities of MWCNT to the mesothelial cells and screen the changes of microRNA profile after exposure to MWCNT.Methods:A LDH method was used to test the cytotoxicities of MWCNT to MeT-5A cell lines. And then the differentially expressed miRNAs between mesothelioma cells and normal mesothelial cells were selected from previous work of research group. Among the significant expression changed miRNAs, 5 were verified by RT-qPCR in mesothelioma cells. The same five ones were further tested in MeT-5A cells exposed to 10 μg/cm2MWCNT for 8, 24, 48, 72 h by RT-qPCR. Target genes of 5 miRNAs were predicted using Targetscan and miRanda softwares. David6.7 was used to perform GO enrichment and KEGG pathway analysis of target genes. All the data were analyzed by one-way ANOVA and Dunnett-T test in SPSS17.0.Results:After 24 h exposure to MWCNT, cell proliferation was significantly suppressed at more than 20 μg/cm2concentration. Among the differentially expressed miRNAs, 5 were chosen to further vestified, namely hsa-miR-155 (up-regulated) , hsa-miR-30 d-5p, hsa-miR-34c-5p, hsa-miR-28-5p and hsa-miR-324-5p (down-regulated) , which were consistent with the miRNA array results. The 5 miRNAs also had the same expression changes in MeT-5A cells after exposure to 10 μg/cm2MWCNT for different time periods. The potential target genes of the 5 miRNAs may be AKAP13, CCND3, Twist and E-Cadherin, which mainly involved in TGF-β signal pathway, small cell lung cancer, etc.Conclusion:MWCNT could induce to MeT-5A cells, and also cause miRNA expression changes. The differential changed miRNAs may involve in cancer related signal pathways.
Institute of occupational diseases prevention and control, Zhejiang Academy of Medical Sciences, Hangzhou, Zhejiang, 310013, China.
X H
XH
Yin
L
L
Ju
W
W
Wu
Y
Y
Xiao
S B
SB
Ying
M
M
Zhang
J L
JL
Lou
Z Y
ZY
Jia
H L
HL
Xia
L J
LJ
Zhu
X
X
Zhang
D015703:Antigens, CD; D015820:Cadherins; D002460:Cell Line; D049109:Cell Proliferation; D004847:Epithelial Cells; D020869:Gene Expression Profiling; D006801:Humans; D035683:MicroRNAs / Q000235:genetics*; D037742:Nanotubes, Carbon / Q000633:toxicity*; D012984:Software; D016212:Transforming Growth Factor beta
D015703:Antigens, CD; C513384:CDH1 protein, human; D015820:Cadherins; C570628:MIRN28 microRNA, human; C000599310:MIRN324 microRNA, human; D035683:MicroRNAs; D037742:Nanotubes, Carbon; D016212:Transforming Growth Factor beta