The Mechanism Of Isoliquiritigenin Induced Apoptosis In Human Hepatocellular Carcinoma Cells

Hepatocellular carcinoma(HCC)is one of the most common malignant tumors and the third cause of cancer-related deaths.The characters were the fastly development,high difficulty of control and poor postoperative performance,which has serious harm to human health and life.Most of the commonly used anti-liver cancer drugs have many problems,such as strong side effects,high price and poor targeting.However the natural medicine can not only alleviate the various clinicopathological characteristics,but also increase the survival time of patients.So it has become an urgent problem to search an anti-liver cancer drug.Isoliquiritigenin(ISL),a natural flavonoid extracted and isolated from plant licorice,has shown various pharmacological activities,including anti-inflammatory,antioxidant and anticancer.In this study,we used liver cancer cells as models and investigate the anticancer mechanism of ISL.And then we evaluated the effects of ISL on HCC cell proliferation,apoptosis induction,cell cycle arrest and ROS generation and its related signal transduction pathways,and provide the theoretical basis for the treatment of cancer and the development of new drugs.In this study,the killing effect of ISL on liver cancer cells was detected by CCK-8.The apoptotic induction of ISL on liver cancer cells was investigated by Hochest/PI staining,Annexin V/PI double stainning and flow cytometry.The regulation of ISL on the downstream apoptotic proteins,cell cycle arrest and cycle proteins were analyzed by flow cytometry and western blot in HCC cells.The levels of MAPK/STAT3/NF-κB pathways and nuclear transcription factors STAT3 and NF-κB,the relationship between MAPK and STAT3 pathways were evaluated by western blot.The regulation effects of ISL on reactive oxygen species(ROS)and related signaling pathways in HCC cells were evistigated by DCFH-DA staining.Compared with the 5-FU group,ISL markedly inhibited the proliferation of Hep G2 and Hep3 B cells.And ISL had no obvious cytotoxic effects on L-02 and QSG-7701 cells.ISL also induced mitochondrial dependent apoptosis by up-regulating the expression of Bax,cle-caspase-3 and cle-PARP,and down-regulated the expression of Bcl-2 in Hep G2 cells.ISL induced G2/M cell cycle arrest by increasing the expression of p21,decreasing the expression of p27,Cyclin B1 and CDK1/2.Moreover,ISL could inhibit the expression of p-ERK,p-STAT3 and NF-κB,while promoted the expression level of p-JNK,p-p38 and IκB-α.ISL could inhibit the expression of STAT3 and NF-κB nuclear transcription factor.ISL could also induce apoptosis by increasing the generation of ROS in Hep G2,and these effects were blocked by NAC.In summary,ISL has a good killing effect on two liver cancer cells,and the molecular mechanisms of ISL-induced G2/M cell cycle arrest and caspase-dependent apoptosis was by promoting the generation of ROS and subsequently regulating the expression of MAPK/STAT3/NF-κB pathway-related proteins,leading to inhibition of HCC cells growth.