**4.1. Traditional Chinese medicines and its active compounds for inhibiting the proliferation and inducing cell death in liver cancer cells**

Jie Du Xiao Zheng Yin (JDXZY) is a typical TCM formula for liver cancer treatment. Recent studies have found that JDXZY can suppress the proliferation of liver cancer cells in a dose‐ and time‐dependent manner [30]. *In vivo* studies also confirmed that JDXZY could increase the tumor apoptotic index and reduce tumor size in nude mice. Mechanism studies showed that JDXZY may induce human hepatoma HepG2 cell apoptosis by mitochondrial‐related pathway. Treatment with JDXZY can activate caspase‐9 and caspase‐3 as well as up‐regulate the ratio of Bax/Bcl‐2, which may partially demonstrate its antihepatoma effects. In addition, another study also found that JDXZY could inhibit the proliferation of HepG2 cells by arresting the cell cycle at the G0/G1 phase [31].

Bufalin is a digoxin‐like agent isolated from Chansu which is a precious CMs extracted from parotoid glands of toads. Previous studies found that bufalin exhibited significant anticancer activities in many cancer cell lines *in vitro* and *in vivo*. A recent research showed that bufalin can suppress AKT‐related signaling pathway in HepG2 and HCCLM3 cell lines. After bufalin treatment, the proliferation‐related proteins in AKT/GSK3beta/beta‐catenin/E‐cadherin‐ signaling pathway were significantly inhibited [32]. Thus, bufalin may be a potential anti‐ hepatoma agent by inhibiting the proliferation of cancer cell.

*Gekko swinhonis* Guenther also called ShouGong in TCM; it was a widely used anticancer drug in China for hundreds of years. Modern science has demonstrated that one of the major active components in *Gekko swinhonis* Guenther is sulfated polysaccharide–protein complex (GSPP). Previous studies showed that GSPP can suppress the proliferation of liver cancer cells and arrest the cell cycle in SMMC‐7721 cells with little direct toxicity [33]. In addition, Gepsin, the monomeric protein in GSPP also showed strong inhibition on the proliferation of hepatocar‐ cinoma Bel‐7402 cells [34].

Many researches have revealed the strong anticancer activity of berberine. The protective effects of berberine on liver diseases including liver cancer have been widely studied by our groups. The quaternary ammonium salt derived from Coptidis Rhizome (Huanglian in TCM) can induce both autophagy and apoptosis in hepatocellular carcinoma cells (HCC). Further mechanisms studies demonstrated that berberine could induce mitochondrial apoptotic signaling pathway in MHCC97‐L and HepG2 cells via upregulating Bax expression. Further‐ more, berberine could also induce autophagic cell death in HCC cells through inhibiting the activity of Akt and activating the p38 MAPK‐signaling pathway [35].
