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. 2019 Apr 23;30(17):1983–1998. doi: 10.1089/ars.2017.7486

FIG. 6.

FIG. 6.

C23 inhibits HG-induced cancer cell progression via PP2Cd inhibition mediated by HSP27 induction. (A) MCF-7 cells were either untreated or subjected to HG challenge in the presence or absence of CCT007093 or C23, and cell lysates underwent immunoblotting for the proteins as indicated. (B) The intracellular oxidative stress was detected by the assay of carbonyl groups in proteins. *p < 0.05 versus control; #p < 0.05 versus HG. (C) MCF-7 cells were transiently transfected with 0.5 μg of pG13-LUC reporter plasmid. About 6 h after transfection, cells were treated as (A). Luciferase activity was determined from the transfected cell extracts. Values (mean ± SD) are expressed as fold over untreated control. *p < 0.05 versus control; #p < 0.05 versus CCT007093; $p < 0.05 versus HG+CCT007093. (D) MCF-7 cells were transfected with Control siRNA or HSP27 siRNA for 24 h followed by incubation with HG or HG/C23 combination for 24 h. Cell lysates underwent immunoblotting for the proteins as indicated. Uncropped blots are shown in Supplementary Figure S10. *p < 0.05 versus control; #p < 0.05 versus HG/Control siRNA; $p < 0.05 versus control/HSP27 siRNA; ♣p < 0.05 versus HG/Comp23/Control siRNA. The intracellular oxidative stress (E) and p53 transcription activity (F) were detected by the assay of carbonyl groups in proteins and luciferase reporter gene (PG13-Luc) assay from cells treated as (D). *p < 0.05 versus control; #p < 0.05 versus HG/Control siRNA; $p < 0.05 versus HG/Comp23/Control siRNA. (G) MCF-7 cells transfected with Control siRNA or HSP27 siRNA were incubated with HG or HG/C23 combination for 48 h, and subjected to wound-healing assay. MTT cell proliferation assay (H) and in vitro invasion assay (I) were performed on MCF-7 cells treated as (G). *p < 0.05 versus control; #p < 0.05 versus HG/Control siRNA; $p < 0.05 versus HG/Comp23/Control siRNA. HSP27, heat shock protein 27. Color images are available online.