Abstract:

Objective To explore the targeting relationship between miR-150-5p and non-SMC condensate Ⅰ complex subunit G （NCAPG） and its effect on the biological function of hepatocellular carcinoma （HCC） cells， and to provide the potential targets for the clinical diagnosis and treatment of HCC. Methods The targeting relationship between miR-150-5p and NCAPG was verified by dual-luciferase reporter gene experiment. The Huh7 cells were divided into mimic negative control （mimic NC）group， miR-150-5p mimic group， miR-150-5p mimic+overexpression negative control （miR-150-5p mimic+ovNC）group and miR-150-5p mimic+overexpression NCAPG （miR-150-5p mimic+ovNCAPG） group. The expression levels of NCAPG mRNA in the cells in various groups were detected by real-time fluorescence quantitative PCR （RT-qPCR） method， and the expression levels of NCAPG protein in the cells in various groups were detected by Western blotting method； 5-ethynyl-2'-deoxyuridine （EdU） assay was used to detect the EdU positive rates of cells in various groups， flow cytometry was used to detect the apoptotic rates， and the numbers of migration and invasion cells were detected by Transwell assay. The nude mice were divided into agomiR-NC group， agomiR-150-5p group， agomiR-150-5p+overexpression negative control（agomiR-150-5p+ovNC） group and agomiR-150-5p+overexpression NCAPG （agomiR-150-5p+ovNCAPG） group. The Huh7 cells formed subcutaneous tumors， and the tumor volumes and tumor weights of nude mice in various groups were measured. Results The dual luciferase reporter gene assay results confirmed that NCAPG was the target gene of miR-150-5p. Compared with mimic NC group， the expression levels of NCAPG mRNA and protein and the EdU positive rate in the Huh7 cells in miR-150-5p mimic group were significantly decreased （P<0.05），the apoptotic rate was significantly increased （P<0.05），and the numbers of migration and invasion cells were significantly decreased（P<0.05）； compared with miR-150-5p mimic +ovNC group， the expression levels of NCAPG mRNA and protein and the EdU positive rate in the Huh7 cells in miR-150-5p mimic +ovNCAPG group were significantly increased （P<0.05），the apoptotic rate was significantly decreased （P<0.05），and the numbers of migration and invasion cells were significantly increased（P<0.05）. Compared with agomiR-NC group， the transplanted tumor volume and tumor weight of the nude mice in agomiR-150-5p group were significantly reduced （P<0.05）； compared with agomiR-150-5p+ovNC group， the transplanted tumor volume and tumor weight of the nude mice in agomiR-150-5p ovNCAPG group were significantly increased （P<0.05）. Conclusion MiR-150-5p inhibits the HCC cell proliferation， migration， invasion and tumor growth through targeting binding and inhibitiing the NCAPG expression， and promotes the apoptosis.

Key words: Non-SMC condensate Ⅰ complex subunit G, Hepatocellular carcinoma, MiR-150-5p, Cell proliferation, Cell migration, Cell invasion