Multi-Target Inhibition of Hepatocellular Carcinoma via DIBP from Glycyrrhiza uralensis: A Systems Biology and Experimental Approach

Hepatocellular carcinoma (HCC) is one of the major causes of cancer-related deaths world wide. Its aggressive nature and late diagnosis imply a very poor survival rate. Therefore there is dire necessity to find new multi-target therapies. In this study, we have identified five veritable hub genes in HCC: CDK1, FOXM1, PTTG1, STMMN1 (overexpressed), and IGFBP3 (downregulated). Through integrated bioinformatics, we have established the clinical relevancy of these five hub genes. The genes are mainly involved in the mitotic processes (e.g. organelle fission, nuclear division, chromosome segregation, etc.), and are over-represented in cell cycle, p53, MAPK, TGF-beta, Hippo signaling. Overexpression of CDK1, FOXM1, PTTG1 and STMN1 is linked with survival and low IGFBP3 to adverse outcomes. The results of immune infiltration showed that the STMN1 level is especially associated with tumor immune cells. Diisobutyl phthalate (DIBP) is one of the bioactive compounds of Gancao, (Glycyrrhiza uralensis Fisch) which we tested by molecular docking, MD simulation, DFT, ADMET prediction, MM/GBSA, and MTT cytotoxicity assays. DIBP binds each hub protein with a stable affinity in silico, exhibits optically favorable drug-like properties and pharmacokinetics, and cause dose-dependent cell death of HCC cell line in in vitro studies. Such in vitro findings compare to the reported selective cell killing of cancer cells by DIBP. In conclusion, the identified hub genes can be used as important diagnostic and prognostic indicators in HCC, and DIBP can serve as a multi-target non-toxic agent against HCC that requires further in vitro research.