Screening And Mechanism Research Of Anti-tumor Cell Invasion And Migration Small Molecule Drugs

Despite recent advances in the treatment of cancer,metastatic disease remains largely incurable and the main cause of cancer-related deaths.To combat the risk for eventual metastasis,many patients are over-treated with the intent of preventing dissemination of their disease.Therapies that specifically target the motility of tumor cells could significantly improve cancer treatment by removing the threat of systemic disease and decreasing the dependency on therapeutics with detrimental side-effects.E-cadherin is a transmembrane protein that maintains intercellular contacts and cell polarity in epithelial tissue.Epithelial-mesenchymal transition(EMT)refers to the process of converting epithelial cells with polar polarity into motile mesenchymal cells.The down-regulation of E-cadherin contributes to the induction of the epithelial-to-mesenchymal transition(EMT),resulting in an increased potential for cellular invasion of surrounding tissues and entry into the bloodstream.E-cadherin,as an EMT epithelial marker,plays an important role in the acquisition of tumor cell invasion and migration ability during cancer progression.A large number of reports link the loss of E-cadherin expression with invasive,dedifferentiated,poorly differentiated cancers.Loss of E-cadherin has been observed in a variety of human tumors as a result of somatic mutations,chromosomal deletions,silencing of the CDH1 gene promoter,and proteolytic cleavage.So far,there are no drugs that directly target E-cadherin expression.Loss of E-cadherin,which is caused by several tumor promoting factors,is associated with metastasis and poor prognosis in many neoplasms.In this study,we aimed to identify small molecule compounds that restore the expression of E-cadherin,because these molecules are most likely to suppress tumor malignancy by restoring E-cadherin function and/or by inhibiting signals that suppress E-cadherin expression.Therefore,this experiment is based on E-cadherin as a target by In-Cell Western high-throughput screening technology from the protein level screening for tumor cell invasion and migration ability of the small molecule compounds significantly inhibited.First,the expression of E-cadherin in various cancer cells was detected.E-cadherin low expression Hep G2 hepatoma cells were selected from 334 small molecular compounds for screening.The preliminary screening data analysis selected the first 11 small molecules that induced E-cadherin expression to varying degrees.Subsequent screening and validation of the compounds;detection of the immunofluorescence levels and protein levels of the four small molecule compounds with more pronounced effects;the two small molecule compounds AW01178(No.291)and AC36050(No.287)with obvious effects were performed One-step function exploration.The morphologically discovered small molecule compounds screened transformed the highly metastatic triple-negative breast cancer cells MDA-MB-231 cells from a mesenchymal-like squamous epithelium;afterwards,scratch experiments,immunofluorescence experiments,and transwell experiments were performed.It was found that screening and obtaining small molecule compounds can significantly inhibit the invasion and metastasis of MDA-MB-231 cells.At the same time,using MTT experiments we found that there is no significant cell viability in non-tumor cells MCF10 A.Finally,we explored the mechanism of E-cadherin expression induced by small molecule compounds and found that it can significantly up-regulate the level of histone H3 acetylation,but it does not affect the level of H3 methylation.Taken together,our experiments using a high-throughput screening system to screen out the ability to significantly inhibit the invasion and migration of tumor cells with little effect on non-tumor cell viability.This later provided more clues on the development and utilization of cancer treatment drugs,small molecule drug modification,and related mechanism research.