Fig. 1.

The effects of PB2 on the proliferation, cell cycle, and apoptosis of HCC in vitro and in vivo. a The cell viability was determined using the CCK8 assay after PB2 treatment for 24, 48, and 72 h. b The colony formation of HCC-LM3 and SMMC-7721 cells. c Cell cycle distribution. PB2 caused cell cycle arrest in the S phase. d Flow cytometry analysis of apoptosis. e Hoechst 33328 staining of HCC cells (original magnification, 200×). The apoptotic cells were stained with bright blue fragments and fluorescence in the nucleus. f Western blot analysis of proliferation- and apoptosis-related markers. g The gross manifestation and volumes of tumors (n = 5). h The hematoxylin and eosin (H&E) staining and TUNEL staining of tumor sections in both NC and PB2 groups (original magnification, 100× or 200×). There were many cancer cells and neo-vessels diffused in the solid neoplastic tissue in the NC group, while there were more lesions of necrosis and less neo-vessels in the PB2 group. In TUNEL staining, apoptotic cells were indicated by red arrows. i The H&E staining of heart, kidney, and lung from the NC and PB2 groups (original magnification, 200×). PB2 treatment for a month did not harm these organs. *P < 0.05 vs. the NC group (1c: one-way ANOVA; 1d&1d: Student’s t-test)