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Fig. 4 | Molecular Medicine

Fig. 4

From: Nanocell-mediated delivery of miR-34a counteracts temozolomide resistance in glioblastoma

Fig. 4

Down-regulation of multiple proteins by miR-34a contributes to sensitization to TMZ. a miR-34a reduces protein levels of ATM, Bcl2, EGFR, UGCG and cMET. Representative western blot images show reduction in ATM, Bcl2, EGFR, UGCG and cMET protein levels in glioblastoma cells. Glioblastoma cells were transfected with 30 nM miR-34a or control miRNA, and cells were lysed 48 h later for protein analysis by western blotting. b Bcl2 siRNA reduces Bcl2 protein levels. Representative western blot images show reduction in Bcl2 protein levels in A172 glioblastoma cell line. A172 cells were transfected with two different Bcl2 siRNA or control siRNA at a concentration of 30 nM, and cells were lysed 48 h later for protein analysis by western blotting. c Inhibition of Bcl2 or cMet sensitizes to temozolomide. A172 cells were reverse-transfected with two different Bcl2 siRNAs, miR-34a and control miRNA 48 h prior to TMZ therapy or treated with JNJ-38877605 2 h prior to being treated with different concentrations of TMZ. Cell proliferation was determined by SRB. All data represent mean SRB values (± SD) from three independent experiments. d miR-34a transfection does not alter MGMT levels. Representative western blot images show reduction in cMET, being used as a positive control, while MGMT protein levels are not affected by miR-34a transfection in the T98G and GBM6 glioblastoma cell cultures. T98G cells were transfected and analyzed similarly to A172 cells in (A). GBM6 cells were transduced with a lentivirus, overexpressing miR-34a only upon doxycycline induction. miR-34a expression was induced in GBM6 cultures for 72 h after which cells were lysed for protein extraction

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