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. 2017 Sep 15;14(5):6285–6290. doi: 10.3892/ol.2017.6965

Figure 2.

Figure 2.

Gemcitabine-induced upregulation of HMGB1 contributes to autophagy in HIPC cells. (A) PC-3 cells were treated with 2, 4, 8, 16 or 32 µg/ml of gemcitabine for 48 h. The expression levels of HMGB1 were analyzed by western blot analysis. (B) HMGB1 mRNA levels were measured using qPCR. All experiments were independently repeated at least three times. *P<0.05. (C and D) PC3 cells were transfected with shRNA or pcDNA3.1-HMGB1 and screened with G418. The PC3 cells transfected with shRNA were treated with 10 µg/ml gemcitabine for 48 h. The expression levels of indicated proteins were analyzed by western blot analysis. (E and F) The transfected PC3 cells were treated with 10 µg/ml gemcitabine or DMSO for 48 h. HSPB1 and Beclin-1mRNA levels were measured with qPCR. All experiments were independently repeated at least three times. *P<0.05. qPCR, quantitative polymerase chain reaction; Gem, gemcitabine; Ctrl, control; LC3, light chain 3; SQSTM1, sequestosome 1; HMGB1, high mobility group box1; HSPB1, heat shock protein β-1; shCtrl, control short hairpin RNA; shHMGB1, short hairpin RNA against HMGB1.