Figure 8.

Gefitinib-induced hepatotoxicity is alleviated by PLK1 inhibitor BI-2536 without compromising the anti-cancer activity of gefitinib. (A) HL-7702 cells were treated with BI-2536 or gefitinib or vehicle for 24 h. The expressions levels of LC3 and COX6A1 were determined. HL-7702 cells were treated with BI-2536 or gefitinib or vehicle for 48 h. Apoptotic cells were identified by flow cytometric analysis (n = 3, one-way ANOVA, LSD test). (B–E) Female ICR mice (n = 6 per group) were treated with BI-2536 (5 mg/kg, twice a week) by intraperitoneal injection or gefitinib (200 mg/kg/day) by gavage. (B) Liver lysates were used to detect the expressions of COX6A1 and LC3. (C) Relative liver weights were calculated (n = 6, one-way ANOVA, LSD test). (D) Serum samples were collected to analyze the levels of GPT/ALT and GOT1/AST (n = 6, one-way ANOVA, Dunnett T3 test). (E) Liver tissues from the mice were collected and stained with H&E for histopathological analysis. Yellow arrowheads indicated the cellular level showing prominent hematoxylin and eosin staining objects that were suggestive of apoptosis in specific regions. For 100× magnification, scale bar: 100 µm; for 200× magnification, scale bar: 50 µm. (F) SRB staining analysis was carried out to determine the survival rates of three different NSCLC cell lines (n = 3, one-way ANOVA, LSD test). Western blot was repeated at least three times and densitometric analysis was carried out. The data are expressed as the mean ± SD; n.s = no significance; *p < 0.05; **p < 0.01; ***p < 0.001. Abbreviations: GEFI, gefitinib; LW, liver weight; BW, body weight; GOT1/AST, glutamic-oxaloacetic transaminase 1, soluble; GPT/ALT, glutamic pyruvic transaminase, soluble; H&E, hematoxylin and eosin; SRB, sulforhodamine B; SE, short exposure; LE, long exposure