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Fig. 2 | Journal of Experimental & Clinical Cancer Research

Fig. 2

From: HSP90AA1-mediated autophagy promotes drug resistance in osteosarcoma

Fig. 2

HSP90AA1 reduces osteosarcoma cells sensitivity to chemotherapy by decreasing apoptosis. a MG-63 and U-2 OS osteosarcoma cells were transfected with control shRNA and HSP90AA1 shRNA for 48 h. Then the expression levels of HSP90AA1 were analyzed by Western blot and quantitative real-time PCR (n = 3; *, p < 0.05 versus control shRNA group). b, c and d, after transfection with control shRNA and HSP90AA1 shRNA, MG-63 and U-2 OS cells were treated with Cis (20 μmol/L), Dox (0.2 μg/mL) and Mtx (50 μmol/L) for 24 h. Then cell viability was determined at 12, 24 and 48 h by CCK-8 assay (b). Apoptosis was analyzed at 24 h by flow cytometric analysis of Annexin V-PE/PI staining (c; n = 3; *, p < 0.05 versus control shRNA group) and both cleaved and total PARP in MG-63 cells were analyzed by Western blot (d). e After transfection with control shRNA and HSP90AA1 shRNA, MG-63 and U-2 OS cells were exposed to Cis (20 μmol/L), Dox (0.2 μg/mL) and Mtx (50 μmol/L) for 24 h in the presence or absence of ZVAD-FMK (20 μmol/L). Then apoptosis was evaluated by Colorimetric Caspase 3 Assay Kit (n = 3; *, p < 0.05 versus control shRNA group). f MG-63 and U-2 OS cells were infected with control (pHBLV control) and HSP90AA1-expressing lentiviruses (pHBLV HSP90AA1). The protein level of HSP90AA1 was assayed by Western blot. G and H, HSP90AA1-overexpressing MG-63 and U-2 OS cells were treated with Cis (20 μmol/L), Dox (0.2 μg/mL) and Mtx (50 μmol/L) for 24 h. Cell viability was determined at 12, 24 and 48 h by CCK-8 assay (g) and apoptosis was analyzed at 24 h by measuring Annexin V-PE positive cells by flowcytometry (h; n = 3; *, p < 0.05 versus control shRNA group)

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