Abstract:
Objective To explore the effect of RNA interference (RNAi) plasmid on proliferation, apoptosis of status of Panc-1 cells by silencing oncogene AKT2, and investigate its possible mechanism.
Methods Pancreatic cancer cell line Panc-1 was applied to constructe the RNAi plasmid-targeting oncogene AKT2 and to transfect cells transiently and stably. The proliferation status was determined using CCK-8 method and soft agar colone formation test. The apotosis status of the cancer cells in vitro was determined using Heochst and Annexin V-FITC/PI methods. The protein level of AKT2 and caspase-3 kinase were detected using Western blotting. Finally, we evaluate the in vivo effect of the recombinant plamids on Panc-1 cells.
Results By silencing the oncogenes AKT2, RNAi plasmid effectively down-regulate the mRNA and protein levels of of AKT2 in Panc-1 cells reduced cell proliferation and colony formation of Panc-1 cells, induced apoptosis in Panc-1 cells, increased the protein level of caspase-3 protein, and inhibited tumor growth in vivo.
Conclusions RNAi can inhibit the expression of oncogene AKT2 and therefore effectively inhibit the growth of pancreatic cancer cells and promote their apoptosis. Gene therapy targeting AKT2 may be a promising target for pancreatic cancer.
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