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Background: Esophageal squamous cell carcinoma (ESCC) is among one of the leading causes of cancer death worldwide owing to late detection and low survival rate. The clinical outcome of ESCC remains dismal. To date, the disease lacks available targeted therapies. Recently, drugs approved by the Food and Drug Administration have been reported to have potential as cancer chemoprevention agents. Methods: Benzydamine, available as a hydrochloride salt, a locally acting non-steroidal anti-inflammatory drug, was screened out among FDA-approved drugs owing to its effective cytotoxic effect on KYSE450 cells, which remained unexplored. Mass spectrometry, kinase prediction and Swiss Target Prediction were used to verify the potential target(s) of benzydamine. Patient-derived oesophageal xenograft mouse model were used to investigate the effect of benzydamine on tumor growth in vivo.Results: We found that benzydamine inhibited anchorage-dependent and -independent growth of ESCC cells. Kyoto Encyclopedia of Genes and Genomes pathway enrichment revealed that benzydamine attenuated five signaling pathways, including the DNA replication pathway. We further found that benzydamine could bind to CDK2 in its ATP-binding site. Inhibition of the activity of CDK2 suppressed the growth of ESCC cells and led to a G1/S cell cycle arrest. Additionally, knocking-down CDK2 decreased the sensitivity of ESCC cells to benzydamine hydrochloride. Notably, benzydamine suppressed tumour growth in a patient-derived oesophageal xenograft mouse model of ESCC in vivo.Conclusions: We have identified CDK2 as a potential target of benzydamine for the treatment and prevention of ESCC. Benzydamine suppressed the growth of ESCC cells by inhibiting the activity of CDK2.

Background Esophageal squamous cell carcinoma (ESCC) is a fatal disease with poor prognosis. The predominant reason for ESCC-related death is distal metastasis. A comprehensive understanding of the molecular mechanism underlying metastasis is needed for improving patient prognosis. T-LAK cell-originated protein kinase (TOPK) is a MAPKK-like kinase, which plays a vital role in various physiological and pathophysiological processes. However, the role of TOPK in ESCC metastasis is unclear. Methods Tissue array was used to evaluate the correlation between TOPK expression and ESCC lymph node metastasis. Wound healing assay, transwell assay, and lung metastasis mice model were used to examine the role of TOPK in the migration of ESCC cells in vitro and in vivo. Protein kinase array, mass spectrometry (MS), and molecular modeling were used to examine the pathways and direct target proteins of TOPK that are involved in ESCC metastasis. Additionally, immunofluorescence and western blotting analyses were performed to verify these findings. Results The enhanced expression of TOPK was correlated with lymph node metastasis in the ESCC tissues. TOPK knockdown or treatment with the TOPK inhibitor (HI-TOPK-032) decreased the invasion and migration of ESCC cells in vitro. HI-TOPK-032 also inhibited the lung metastasis in ESCC cell xenograft in vivo model. Moreover, TOPK promoted the invasion of ESCC cells by activating the Src/GSK3β/STAT3 and ERK signaling pathways via γ-catenin. Conclusion The findings of this study reveal that TOPK is involved in ESCC metastasis and promoted the ESCC cell mobility by activating the Src/GSK3β/STAT3 and ERK signaling pathways. This indicated that TOPK may be a potential molecular therapeutic target for ESCC metastasis. Keywords esophageal squamous cell carcinoma; TOPK; tumor metastasis; Src/GSK3β/STAT3; γ-catenin

Background Esophageal squamous cell carcinoma (ESCC) is a deadly disease with the poor prognosis in the world. The distal metastasis is the most death reason of ESCC. It is needed to have a comprehensive understanding of the molecular mechanism of metastasis to increase the free survive rate. T-LAK cell-originated protein kinase (TOPK) which is a MAPKK-like kinase takes an vital role in many physical and pathophysiological progress. However, the function of TOPK in ESCC metastasis was unclear. Methods Tissue array was used to evaluate the relationship between TOPK and ESCC patient with lymph node metastasis. Wound healing assay, transwell assay and lung metastasis mice model were assessed for the role of TOPK in the migration of ESCC cells in vitro and in vivo respectively. Protein kinase array, MS and molecular modeling were carried out to find the relational pathways and target protein of TOPK. Even, immune-fluorescence and western blot were performed to evaluate the mechanism of TOPK. Results We found that the high level of TOPK was correlated with the aggressive phenotype in ESCC tissues. Knocking down TOPK inhibited the invasion and migration of ESCC cells. We also verified that TOPK inhibitor HI-TOPK-032 inhibited the lung metastasis in ESCC cell exnograft model. Even more, molecular investigation indicated that TOPK promoted the invasion of ESCC cells by activing Src/GSK3β/STAT3, ERK pathway by binding with γ-catenin. Conclusion These findings reveal that TOPK was sincerely related with ESCC cell metastasis and TOPK promoted the invasion of ESCC cells by activing Src/GSK3β/STAT3, ERK pathway. This means that TOPK may be a potential molecular target for ESCC in clinic.