Effect of silencing S-phase kinase-associated protein 2 on chemosensitivity to temozolomide of human glioma cells U251

  • Am J Transl Res. 2019 Apr 15;11(4):2470-2476.
Yue Liu  1 ,  Zhen-Gang Tang  1 ,  Jian-Quan Yang  1 ,  Yi Zhou  1 ,  Yi Lin  1 ,  Wei Lv  1 ,  Guo-Bin Wang  1 ,  Cai-Li Li  1
Affiliations
  • 1. Department of Neurosurgery, Renmin Hospital, Hubei University of Medicine Shiyan, Hubei, P.R. China.
PMID: 31105854
Abstract

Objective: To examine the effect of silencing SKP2 on chemosensitivity of human glioma cells U251 to temozolomide (TMZ).

Methods: Adenoviruses harbouring shRNA targeting SKP2 (i.e. Ad-shSKP2) and non-targeting scrambled shRNA (i.e. Ad-shNC) were used to infect U251 cells. The transduced cells were then treated with TMZ. Cell viability after treatment was assayed using CCK8; while cell cycle and Apoptosis were examined using flow cytometry. To study the effect of silencing SKP2 on Autophagy in U251, we co-transduced the cells with Ad-mRFP-LC3 and Ad-shSKP2/Ad-shNC. The expression of Autophagy marker LC3 after TMZ treatment was studied using microscopy and Western blotting assays.

Results: The cytotoxicity of TMZ (i.e. 20-100 µM) was more significantly seen in Ad-shSKP2-transduced U251 cells than in the Ad-shNC-transduced U251 cells. The IC50 values in shSKP2-U251 were significantly lower than those of the shNC-U251 (P < 0.05). Both TMZ and Ad-shSKP2 alone increased Apoptosis and promoted expression of LC3 in U251. Combined treatment of Ad-shSKP2 and TMZ further elevated Apoptosis and LC3 expression.

Conclusion: Silencing SKP2 in U251 cells increased chemosensitivity to TMZ that was accompanied with enhanced Apoptosis and Autophagy. Targeting SKP2 may be a potential approach to potentiate TMZ treatment in patients with glioma.

Keywords
Glioma; S-phase kinase-associated protein 2 (SKP2); autophagy; chemosensitivity; temozolomide (TMZ).
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