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. 2015 Aug 20;8:2199–2208. doi: 10.2147/OTT.S85948

Figure 5.

Figure 5

Fluoxetine enhances TMZ’s antitumoral effect by disrupting NF-κB–MGMT pathway.

Notes: (A) T98G glioma cells were plated into six-well plate and infected with shRNA lentiviruses targeting MGMT or IKKβ. Forty eight hours after lentiviruses infection, Total cell lysate was harvested and used for Western blotting analysis on MGMT, IKKβ, p-NF-κB/p65, and NF-κB/p65. β-actin served as a loading control. (B) Cytotoxicity analysis on TMZ alone or combined with FLT in T98G cells with or without MGMT and IKKβ depletion. The cell viability in no drug control was regarded as 100%. n=3,*P<0.05, no drug control versus TMZ or TMZ+FLT; **P<0.05, TMZ alone versus TMZ+FLT. (C) Constitutive expression of MGMT in T98G glioma cells. T98G glioma cells were plated into six-well plate and infected with MGMT expression lentiviruses. Forty eight hours after lentiviruses infection, total cell lysate was harvested and used for Western blotting analysis on MGMT. β-actin served as a loading control. (D) Cytotoxicity analysis on TMZ alone or combined with FLT in T98G cells with or without constitutive MGMT expression. The cell viability in no drug control was regarded as 100%. n=3, *P<0.05, no drug control versus TMZ or TMZ+FLT; **P<0.05, TMZ alone versus TMZ+FLT.

Abbreviations: EV, empty vector; FLT, fluoxetine; IKKβ, IκB kinase β; MGMT, O6-methylguanine-DNA methyltransferase; TMZ, temozolomide; NF-κB, nuclear factor of kappa light polypeptide gene enhancer in B-Cells; Scr. scramble; shRNA, short hairpin RNA.