Background: Autophagy plays a “double-edged sword” in the process of tumorigenesis, development and metastasis.

Objective: In this study, we explored the effect of PI3K/AKT/mTOR autophagy-related signaling pathway on regulating and controlling the invasion and metastasis of liver cancer cells by Bufalin.

Methods: The cell counting, migration, adhesion and invasion assay were used to evaluate the effect of Bufalin on cell proliferation, invasion and metastasis. The protein expression of PI3K/AKT/ mTOR signaling pathway were detected by the Western Blotting technique.

Results: After inhibiting autophagy of HCC-LM3 cells, the inhibitory effect of Bufalin on adhesion, migration and invasion of HCC-LM3 cells was significantly enhanced. Synergistic inhibition was strongest when different autophagy inhibitors were combined with 3MA and CQ. After inhibiting autophagy, Bufalin significantly inhibited the protein expression of P-AKT, Cyclin D1, MMP- 2, MMP-9 and VEGF in HCC-LM3 cells. The protein expression of PTEN and E-Cadherin in HCC-LM3 cells was significantly increased.

Conclusion: The present study shows that the anti-tumor effect of Bufalin mainly inhibit proliferation, extracellular matrix degradation and angiogenesis of HCC by influencing autophagy. These findings confirm the capability of Bufalin in inhibiting metastasis of HCC and in parallel to current patents, could be applied as a novel therapeutic strategy in the prevention of metastasis of HCC.

背景：自噬在肿瘤的发生、发展和转移过程中扮演着一把“双刃剑”。

目的：本研究探讨PI3K/AKT/mTOR自噬相关信号通路对蟾毒灵调控肝癌细胞侵袭转移的作用。

方法：采用细胞计数、迁移、黏附和侵袭实验评价蟾蜍灵对细胞增殖、侵袭和转移的影响。Western Blotting技术检测PI3K/AKT/mTOR信号通路蛋白表达。

结果：抑制HCC-LM3细胞自噬后，蟾毒灵对HCC-LM3细胞粘附、迁移和侵袭的抑制作用明显增强。当不同的自噬抑制剂与 3MA 和 CQ 联合使用时，协同抑制作用最强。Bufalin抑制自噬后，显着抑制HCC-LM3细胞中P-AKT、Cyclin D1、MMP-2、MMP-9和VEGF的蛋白表达。HCC-LM3细胞中PTEN和E-Cadherin的蛋白表达显着增加。

结论：本研究表明，蟾毒灵的抗肿瘤作用主要通过影响自噬来抑制HCC的增殖、细胞外基质降解和血管生成。这些发现证实了 Bufalin 抑制 HCC 转移的能力，并且与现有专利平行，可作为一种新的治疗策略应用于预防 HCC 转移。