Chinese Clinical Oncology ›› 2018, Vol. 23 ›› Issue (8): 673-679.

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Regulation of VEGFC expression by microRNA507 and its effect on biological behavior and PI3K/Akt pathway of hepatoma cells

  

  1. Department of Oncology, Integrative Medicine, Cancer Hospital Affiliated to Zhengzhou University
  • Online:2018-08-31 Published:2018-09-07

Abstract: ObjectiveTo investigate the effect of microRNA507 (miR507) on the proliferation, apoptosis, migration and PI3K/Akt signaling pathway of hepatoma cells and analyze the targeting regulation of miR507 on vascular endothelial growth factor C (VEGFC). 
MethodsQuantitative realtime polymerase chain reaction (QPCR) was used to detect the levels of miR507 and VEGFC mRNA in the cancer tissues and paracancerous tissues of 90 patients with hepatoma cancer from January 2014 to March 2017. Human hepatoma cells HepG2 were transfected with miR507 mimics (overexpressing group) or negative control (NC group) by liposome method. QPCR was used to detect the miR507 level after 48 htransfection. MTT method was used to compare the cell proliferation at different treatment times (0, 12, 24, 48 h). Flow cytometry with Annexin ⅤFITC/PI staining was used to detect the cell apoptosis, and the scratch test was used to compare the migration of each group. QPCR and Western blotting were employed to detect the mRNA and protein levels of VEGFC and PI3K/Akt signal pathwaysrelated genes including pAkt and pPI3K. Dual luciferase reporter gene test was used to verify the target relationship and binding site between miR507 and target gene VEGFC. 
ResultsQPCR detection showed that the level of miR507 in hepatoma cancer tissues was 0672±0089, lower than 1142±0136 in paracancerous tissues and level of VEGFC mRNA in hepatoma cancer tissues was 1482±0156, higher than 1021±0087 in paracancerous tissues (P<005). Compared with the NC group and the control group, the proliferative activity of HepG2 cells in the overexpressing group was significantly decreased after transfection of 2448 h (P<001). The apoptotic rate of HepG2 cells in overexpressing group was (1726±196)%, higher than (785±087)% of NC group and (813±069)% of control group (P<005). The healing rate of HepG2 cells in overexpressing group was (3965±487)%, lower than (5818±273)% in NC group and (5724±317)% in control group (P<005). The protein and mRNA levels of pAkt, pPI3K and VEGFC in the overexpression group were lower than those in the NC group and the control group (P<005). The double luciferase reporter gene experiment proved that VEGFC was a direct target of miR507. 
ConclusionMiR507 may regulate HepG2 cells proliferation, apoptosis and migration through targeting VEGFC of PI3K/Akt signal pathway, and serve as a potential therapeutic strategy in hepatoma cancer.


Key words: Hepatoma cancer, MicroRNA507, Proliferation, Apoptosis, Migration, Vascular endothelial growth factor C

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