The effect and mechanism of miR196a in HepG2 cell

To explore the role of miR-196a on the regulatory mechanism in hepatocelluar carcinoma.

The antisense RNA of microRNA-196a was synthesized and cloned into the vector. HepG2 cells were infected by inhibiting miR196a vector. The HepG2 cells were divided into miR196a lower expression group, NC group and N group in vitro. The expression of the targets of miR-196a was detected by qPCR. Cell growth was analyzed by cck8 assay. The invasion was detected by transwell method. Apoptosis was detected by annexinV/PI. The P53, caspase-3, HOXB9, HOXB8 mRNA and their protein was detected by qPCR and Western-blot.

(1) The expression level of miR-196a was less than normal (41%). (2) The proliferation of HepG2 was also markedly suppressed in inhibiting miR196a at the 24 h point than normal about 72.29±2.51% (P<0.01). (3) The number of cells that migrated through the chamber of miR196a inhibiting group is less than normal and NC (P<0.01). (4) The cell apoptosis in miR196a inhibiting group is more than NC and normal group (P<0.05). HOXB8 mRNA and protein expression, in HepG2 cell line miR196a inhibiting group is significantly less than normal, NC (P<0.05). Caspase-3 mRNA and protein expression is maximum in three groups (P<0.05). In three groups there was no significant difference in the expression of P53 mRNA and protein and HOXB9mRNA.

Our results demonstrate that miR-196a can effect the proliferation, the apoptosis and migration of HepG2 cell lines by gene HOXB8, caspase-3 regulation. However, there is no correlation between miRNA196a and P53 and HOXB9.