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Influence of Saikosaponin D on the autophagy level of human hepatoma cells and their molecular mechanism

Shan Li, Jianfeng Gao, Jiong Zhang, Fang Chen, Aishe Gao

Purpose: This study aims at studying and discussing the influence of Saikosaponin D (SSD) on the autophagy level of human hepatocarcinoma cells and their molecular mechanism. This study should provide certain theoretical basis for the clinical use of SSD to treat hepatic carcinoma.

Method: This study used in-vitro cell experiments to study the influence of SSD on the clone formation, apoptosis, and proliferation of hepatocarcinoma cells. An electron microscope is then used to examine the influence of SSD on the autophagosome formation in hepatocarcinoma cell, whereas a Western blot method is applied to test the expressions of p-AKT, p-AMPK, and LC3-II in hepatocarcinoma cells.

Results: After an SSD intervention for 48 h, the human hepatocarcinoma cells PLC, MHCC-97H, SMMC-7721, and Huh7 respectively decreased by 19.49 ± 2.92%, 45.41 ± 7.95%, 53.84 ± 5.53%, and 64.04 ± 4.36% compared with those of the control group. After a two-week SSD, the number of cell colonies of human hepatocarcinoma cells PLC, MHCC-97H, SMMC-7721, and Huh7 are obviously reduced when compared with those in the normal control group. The cell apoptosis in human hepatocarcinoma cells is SSD induced, and the apoptosis inductivities of PLC, MHCC-97H, SMMC-7721, and Huh7 are at 11.67 ± 1.17%, 17.54 ± 1.83%, 15.45 ± 1.55%, and 15.54 ± 1.57%, respectively. The autophagosomes in human hepatocarcinoma cells through SSD intervention obviously increased. Western blot displayed that the expression of AKT protein (p-AKT) phosphorylated after SSD processing decreased, whereas the expression of AMPK (p-AMPK) protein increased.

Conclusion: SSD can increase the autophagy of human hepatocarcinoma cells, inhibit the proliferation of carcinoma cells, and promote the apoptosis of cells, which may be attributed to the AMPK/mTOR signal transduction pathway.