Gallbladder cancer (GBC) is a common malignant cancer in the biliary system, which poses a serious threat to human health. It is urgent to explore ideal drugs for the treatment of GBC. Matrine is the main active ingredient of sophora avescentis, with a wide range of biological activities encompassing antiin ammatory, antiviral, immunomodulatory and anti-tumor. However, the underlying mechanism by which Matrine treats GBC is still unclear. The purpose of this study is to investigate the anti-tumor effects of Matrine on GBC in vivo and in vitro, and to clarify the potential regulatory mechanisms. Here, in this primer, we found that Matrine has a signi cant killing effect on GBC through CCK8 and ow cytometry, including arrest of cell cycle, inhibition of GBC cell, and induction of apoptosis. Further studies in vivo con rmed that the inhibitory function of Matrine on tumor growth in NOZ xenografted nude mouse. At the same time, Matrine also signi cantly suppressed the migration and invasion of GBC cells through scratch and Transwell experiments. In addition, by detecting the mRNA and protein levels of epithelialmesenchymal transition (EMT) and matrix metalloproteinases, Matrine furtherly substantiated the suppression of invasion and migration of GBC. From a mechanistic perspective, Matrine effectively decreased the abundance of p-PI3K and p-AKT protein in vivo and in vitro. More importantly, PI3K activator (740 Y-P) antagonized the anti-tumor effect of Matrine, while PI3K inhibitor (LY294002) increased the sensitivity of Matrine for GBC. Based on the above ndings, we conclude that Matrine inhibits the invasion and migration of GBC by regulating PI3K/AKT signaling pathway. Our results indicate the crucial role and regulatory mechanism of Matrine in suppressing the growth of GBC, which provides a theoretical basis for Matrine to be a candidate drug for the treatment and research of GBC.Cell culture GBC-SD cell line, a human GBC cell line, was purchased from Procell (Wuhan, China, Cat. No. CL-0085), while NOZ cell line (Cat. No. MZ-2106) of human GBC and HGBEC cell line (Cat. No. MZ-3197) of human gallbladder epithelial immortalized was both obtained from Ningbo, China. Whelab Bioscience Limited Corporation (Shanghai, China, Cat. No. C1546) provided SGC-996 cell line of the other human GBC. NOZ cells were cultured in DMEM medium containing 10% concentrations of fetal bovine serum (FBS, Wisent, Cat. No. 085-150), 100 U/ml penicillin (Beyotime, Cat. No. C0222), and 100 µg/ml streptomycin (Beyotime, Cat. No. C0222), while SGC-996 and GBC-SD cells were maintained in PRMI 1640 complete medium. The specialized medium was used for culturing HGBEC cells. All cells were cultured at 37 ℃ with 5% carbon dioxide.
CCK8 assayCell viability was evaluated according to Cell Counting Kit-8 (MedChemExpress, Cat. No. HY-K0301) provided by the vender. The cells (HGBEC, GBC-SD, NOZ and SGC-996) were seeded in 96-well plates overnight and treated with different concentrations of Matrine (0-80 µM) for 12-72 hours, or treated with LY294002 (0-8...