Research Paper Volume 13, Issue 18 pp 22286—22297

ASXL1 promotes adrenocortical carcinoma and is associated with chemoresistance to EDP regimen

Silencing ASXL1 sensitized ACC cells to etoposide, doxorubicin and cisplatin (EDP) regimen. (A) Relative firefly-luciferase activity of FSCN1 in NCI-H295R cells where Firefly luciferase activity was normalized to Renilla luciferase activity for all samples to yield relative luciferase activity upon ASXL1 silencing or control; (A, B) Cell count detected using CCK-8 in ACC cell lines with ASXL1 overexpression by adenovirus (Av) or control (AvCON) and/or with FSCN1 silencing by shRNA; (C) Transwell assays used to detect cell invasion with Matrigel in ACC cells with ASXL1 overexpression by adenovirus (Av) and/or with FSCN1 silencing by shRNA, captured at 100×; Flow cytometry used to detect (D) cell cycle profile and (E) apoptosis in ACC cells with ASXL1 overexpression by adenovirus (Av) and/or with FSCN1 silencing by shRNA; Dose-response curves of (F) Doxorubicin, (G) Cisplatin and (H) Etoposide in ACC cells with ASXL1 or FSCN1 silencing by shRNA or control (shCON), with IC50 (μmol) in parentheses (ns = not significant; *P

Figure 4. Silencing ASXL1 sensitized ACC cells to etoposide, doxorubicin and cisplatin (EDP) regimen. (A) Relative firefly-luciferase activity of FSCN1 in NCI-H295R cells where Firefly luciferase activity was normalized to Renilla luciferase activity for all samples to yield relative luciferase activity upon ASXL1 silencing or control; (A, B) Cell count detected using CCK-8 in ACC cell lines with ASXL1 overexpression by adenovirus (Av) or control (AvCON) and/or with FSCN1 silencing by shRNA; (C) Transwell assays used to detect cell invasion with Matrigel in ACC cells with ASXL1 overexpression by adenovirus (Av) and/or with FSCN1 silencing by shRNA, captured at 100×; Flow cytometry used to detect (D) cell cycle profile and (E) apoptosis in ACC cells with ASXL1 overexpression by adenovirus (Av) and/or with FSCN1 silencing by shRNA; Dose-response curves of (F) Doxorubicin, (G) Cisplatin and (H) Etoposide in ACC cells with ASXL1 or FSCN1 silencing by shRNA or control (shCON), with IC50 (μmol) in parentheses (ns = not significant; *P < 0.05; **P < 0.01).