Author/Authors :
Shi, Qingyuan Department of Otorhinolaryngology - HwaMei Hospital - University of Chinese Academy of Sciences (Ningbo No.2 Hospital) - Ningbo - Zhejiang 315010 - P.R.China , Huang, Lizhen Department of Otorhinolaryngology - HwaMei Hospital - University of Chinese Academy of Sciences (Ningbo No.2 Hospital) - Ningbo - Zhejiang 315010 - P.R.China , Dai, Jinhua Department of Clinical Laboratory - HwaMei Hospital - University of Chinese Academy of Sciences (Ningbo No.2 Hospital) - Ningbo - Zhejiang 315010 - P.R.China
Abstract :
microRNA-29 (miR-29) family miRNAs have been mentioned as tumor suppressive genes
in several human cancers. The purpose of this study was to investigate the function of miR-29a in
nasopharyngeal carcinoma (NPC) cells.
Materials and Methods: Human NPC cell line 5-8F was transfected with mimic, inhibitor or scrambled
controls specific for miR-29a. Subsequently, cell viability, migration, apoptosis and expression
changes of VEGF were assessed by trypan blue staining, MTT assay, transwell assay, flow cytometry,
Western blot and RT-qPCR. TargetScan online database was used to predict the targets of miR-29a,
and luciferase reporter assay was carried out for testing the targeting relationship between VEGF and
miR-29a. Western blot analysis was performed to determine the expression changes of core proteins
in PI3K/AKT and JAK/STAT pathways.
Results: Overexpression of miR-29a suppressed 5-8F cells viability and relative migration, but increased
apoptotic cell rate. Consistently, Bcl-2 was downregulated, Bax was upregulated, and caspase-3 and -9
were cleaved by miR-29a overexpression. VEGF was a target gene of miR-29a. Besides, VEGF silence
exerted similar effects like miR-29a, as the viability and migration were repressed and apoptosis was
induced. Finally, we found that PI3K/AKT and JAK/STAT pathways were deactivated by miR-29a or
VEGF silence.
Conclusion: These findings highlighted the tumor suppressive effects of miR-29a on NPC cells, as
its overexpression inhibited 5-8F cells viability, migration, and induced apoptosis. miR-29a exerted
tumor suppressive functions might be via targeting VEGF and deactivating PI3K/AKT and JAK/STAT
pathways.
Keywords :
5-8F cell , miR-29a , Nasopharyngeal carcinoma- (NPC) , PI3K/AKT and JAK/STATpathways , VEGF