Objective: To investigate the effect of RING finger protein 2 (RNF2) knockdown on the biological characteristics and radiosensitivity in glioma U87 cells.
Methods: Plasmids containing shRNA targeting RNF2 were transfected into U87 cells. Real-time quantitative PCR (qRT-PCR) and Western blotting were respectively applied to detect the mRNA and protein level of RNF2. MTT assay was used to detect cell proliferation. Cell cycle and apoptosis were measured by flow cytometry combined with annexin V-FITC/PI staining in the control and RNF2 knockdown cells. Apoptosis was also detected after X-ray radiation.
Results: Both shRNAs efficiently inhibited RNF2 expression in U87 cells. Cell proliferation was obviously depressed in RNF2 knockdown cells. The percentage of cells decreased in S phase (shRNA-NC: 27.31 ± 1.35; shRNF2-1: 16.72 ± 2.90; shRNF2-3: 10.35 ± 1.33) and increased in G1 phase (shRNA-NC: 56.13 ± 1.80; shRNF2-1: 76.32 ± 3.11; shRNF2-3: 80.45 ± 2.83). More cell apoptosis was observed in RNF2 knockdown cells. After X ray radiation, the apoptosis rate was significantly raised in RNF2 knockdown cells (shRNA-NC: 20.88 ± 0.64; shRNF2-1: 39.69 ± 0.57; shRNF2-3: 47.82 ± 0.45).
Conclusion: Knockdown of RNF2 can inhibit cell proliferation, induce cell cycle arrest and promote apoptosis in U87 cells. RNF2 knockdown can obviously increase the sensitivity of U87 cells to X ray radiation.