Lycorine Hydrochloride Suppress the Proliferation, Metastasis and Invasion of Esophageal Cancer by Targeting TRIM22 and Inhibiting the JAK2/STAT3, PI3K/AKT and MAPK/ERK Pathways

Background: Tumor metastasis and poor drug efficacy are two of the prominent causes of therapeutic failure in tumor patients. The underlying molecular mechanism requires further exploration and novel effective curative strategies are urgently needed. Nature is a rich source for novel drug discovery and Lycorine hydrochloride (Lyc.HCL) is a natural alkaloid with tremendous therapeutic potential. However, the molecular mechanisms of anti-tumor are still elusive. In the current study, we investigated the effects and mechanisms of Lyc.HCL against esophageal squamous cell carcinomas (ESCC) that seriously endangers human life. Methods: We detected the expression of tripartite motif containing 22(TRIM22) through immunohistochemistry and western blot. The MTS assay and clone formation assay were used to assess the viability of ESCC cell lines after Lyc.HCL treatment in vitro. Apoptosis and cell cycle regulation was analyzed by flow cytometry. Wound healing and Transwell assays were used to analyze cell migration. Invasion was analyzed by the Matrigel transwell assay. The expressions of janus kinase-2(JAK2)/signal transducer and activator of transcription 3(STAT3), phosphoinositide 3-kinase (PI3K)/protein kinase B(AKT), mitogen-activated protein kinases (MAPK)/extracellular signal-regulated kinase (ERK) pathways were detected through Western blotting. Rescue experiment was performed to underscore the potential target role of TRIM22. In addition, we also explored the vivo anti-ESCC role and mechanism of Lyc.HCL by treating tumor-bearing mice. Results: We found that the TRIM22 protein was highly expressed in ESCC tissues and ESCC cell lines but not in normal esophageal tissue. Lyc.HCL treatment was found to inhibit esophageal squamous cell carcinoma cell proliferation both in vitro and in vivo by blocking the cell cycle to G2 phase, inhibiting cell migration and invasion. Furthermore, Lyc.HCL decreased the expression of TRIM22 and JAK2/STAT3, PI3K/AKT, MAPK/ERK signaling pathways both in vitro and in vivo. Rescue assays showed that overexpressed TRIM22 would promote proliferation, metastasis and invasion, while knocking out TRIM22 significantly reverses these effects. Lyc.HCL suppresses proliferation, metastasis and invasion induced by TRIM22. Observing in animal experiments, depletion TRIM22 delays tumor growth and the effect is significantly reversed by TRIM22 overexpression. Conclusions: Taken together, these findings demonstrate that Lyc.HCL effectively suppresses ESCC both in vitro and in vivo by targeting TRIM22, which is regulated by the JAK2/STAT3, PI3K/AKT and MAPK/ERK pathways, indicating that Lyc.HCL may be the potential novel drug for ESCC and TRIM22 is a considerable target for therapy.