[RNA interference targeting DNA-PKcs inhibits glioma cells malignancies and enhances temozolomide sensitivity].

Objective: To investigate the effect of DNA dependent protein kinase catalytic subunit (DNA-PKcs) on glioma proliferation, invasion and temozolomide sensitivity, and also to explore the potential mechanisms. Methods: Human glioma cell lines H4 and U87 were chosen to carry out RNA interference transfection, and then divided into negative control group (blank group) and siRNA group (test group). The knockdown efficacy of DNA-PKcs siRNA was tested by quantitative PCR and Western blot. The MTS assay and Transwell assay were used to investigate the effect of DNA-PKcs knockdown on glioma cell growth and invasion, respectively. We also used MTS assay to investigate the IC(50) value of temozolomide in negative control group and siRNA groups. Result: Compared with blank group, DNA-PKcs specific siRNA significantly downregulated both mRNA and protein level of DNA-PKcs. MTS assay results demonstrated that 72-hours proliferation of test group were only 52.48%, 54.70% (H4) and 52.98%, 50.45% (U87) of the blank group's counterpart. Transwell assay results showed that the invasiveness abilities of blank and test groups were 1.00±0.03, 0.41±0.05, 0.39±0.04 (H4) and 1.00±0.02, 0.28±0.04, 0.27±0.04 (U87). Moreover, knockdown of DNA-PKcs significantly decreased the temozolomide IC(50) value (H4: 249±27, 97±39, 88±35; U87: 485±41, 86±49, 73±38). Further we applied the Western blot to reveal the mechanism of inhibitory effect of DNA-PKcs knockdown on glioma malignancies and temozolomide sensitivity. We found that downregulation of DNA-PKcs reduced the activity of AKT signal and the expression of its downstream effectors, such as c-Myc, MMP9, and Survivin. Conclusion: RNA interference targeting DNA-PKcs could inhibit glioma malignancies and enhance temozolomide sensitivity. The inhibitory effect of DNA-PKcs knockdown on those biological activities were mainly through inhibition of AKT signal and its downstream effectors.