Figure 6.

Effects of muscone on subcutaneous tumor growth in HepG2 cells. (A and B) Morphology of HepG2 cell subcutaneous tumors in BALB/c nude mice injected with HepG2 cells and treated with muscone (no. 5-8) or DMSO (no. 1-4). (C) Tumor volumes of BALB/c nude mice injected with HepG2 cells and treated with DMSO or muscone. (D) Tumor weight (in mg) of subcutaneous tumors in BALB/c nude mice injected with HepG2 cells and treated with muscone or DMSO. (E) Expression levels of PERK/ATF4/DDIT3 signaling pathway-related proteins (p-eIF2α, p-PERK, ATF4 and DDIT3) and apoptosis-related markers (Bax, Bcl-2 and caspase-3) were detected by western blotting from two different groups of tumor tissues (group 1: no. 1 and no. 5 subcutaneous tumors; group 2: no. 2 and no. 6 subcutaneous tumors). (F) Western blotting was used to analyze the expression levels of autophagy-related markers in two different groups of tumor tissues (group 1: no. 1 and no. 5; group 2: no. 2 and no. 6). Actin was used as a loading control. (G) Reverse transcription-quantitative PCR analysis was used to analyze SESN2 expression levels in HCC tissues (T) compared with corresponding non-cancerous tissues (N). *P<0.05. (H) Western blotting was used to investigate SESN2 expression levels in 5 samples randomly selected from HCC tumor samples compared with corresponding non-cancerous tissue samples. p-eiF2α, phosphorylated eukaryotic initiation factor 2α; p-PERK, phosphorylated protein kinase R-like endoplasmic reticulum kinase; ATF4, anti-activating transcription factor 4; DDIT3, anti-DNA damage inducible transcript 3; p-AMPK, phosphorylated AMP-activated protein kinase; p-mTOR1, phosphorylated mechanistic target of rapamycin kinase 1; SESN2, anti-sestrin 2.