Plumbagin (PL), an active naphthoquinone compound, has been demonstrated to be a potential anticancer agent. However, the underlying anticancer mechanism is not fully understood. In this study, the human hepatocellular carcinoma (HCC) SMMC-7721 cell line was studied in an in vitro model. The cell proliferation was inhibited by PL in a dose-and time-dependent manner.Electron microscopy, acridine orange staining, and immunofluorescence were used to evaluate autophagosome formation and LC3 protein expression in PLtreated SMMC-7721 cells. Real-time polymerase chain reaction and Western blot showed that PL treatment suppressed the expression of apoptosis and autophagy factors (LC3, Beclin1, Atg7, and Atg5), which are associated with tumor apoptosis and autophagy in SMMC-7721 cells. In the study of in vitro tumor nude mouse models, PL can inhibit tumor growth. Cell apoptosis and autophagy of the transplanted tumors were evaluated by hematoxylin and eosin staining, terminal deoxynucleotidyl transferase-mediated dUTP nick endlabeling staining, and Western blot. In addition, in the in vivo studies of HCC cells, we found that pretreatment with the autophagy inhibitor 3-methyladenine blocked the formation of apoptosis induced by PL. In contrast, administration of the apoptosis inhibitor Z-VAD did not affect PL-induced autophagy. Taken together, our findings strongly suggest that PL is a promising drug with significant antitumor activity in HCC. K E Y W O R D S autophagy, Beclin1, hepatocellular carcinoma, microtubule-associated protein 1 light chain, plumbagin J Cell Biochem. 2019;120:9820-9830. wileyonlinelibrary.com/journal/jcb 9820 |

Hepatocellular carcinoma (HCC) is considered as the leading killer disease in the world. So far most of the diagnosis of HCC is mainly established on imaging and biopsy. As sequencing technology is developing quite fast, and it has already been widely applied in the medical area, such as cancer diagnosis. In this article, GSE121248, GSE76427 and GSE60502 datasets were chosen to analyze and screen key genes which could affect the development of liver cancer through the bioinformatics method. The results showed up regulated genes mainly reside in cell division, nucleus, protein binding pathway, and down regulated genes are mostly located in the Oxidation-reduction process, Extracellular region, Heme binding, Metabolic pathway. Secondly, hub gene analysis indicated there were twelve critical hub genes found: RFC4, RACGAP1, CCNB2, CDC20, UBE2C, PTTG1, AURKA, PRC1, NCAPG, CDKN3, TOP2A, KIF20A, AURKA and CDKN3. By applying bioinformatic measures , the genes associated with hepatocellular carcinoma can be efficiently analyzed, that would provide invaluable information for translational studies.