Induced Bladder Cancer Cell Line Death By A Novel Quinazolinyl-bis-aryl Urea Derivative 8g And Its Molecular Mechanism Study

Arylurea derivatives are small molecule multi-kinase inhibitors that have a wide range of biological activities in medicine.Sorafenib（Sora）is a marketed drug that is a typical representation of a uremic group.It is mainly used for the treatment of advanced cancer which cannot be treated with surgical resection.Our group already designed and synthesized a series of arylurea derivatives.During their anti-tumor activity screening,a quinazolyl-bis-aryl urea derivative 8g was found to exhibit good activity against bladder cancer cell lines,but its mechanism of action had not been defined.Therefore,we tried to elucidate the mechanism of 8g at cellular,molecular,and genetic levels.This thesis consists of the following four chapters:In the first chapter,we mainly conducted reviews about arylurea compounds and the mode of death about cancer cells.In the second chapter,its result suggested that 8g had a good anticancer activity against T24 cells(IC₅₀=3.55±0.18μM at 48 h).In addition,its activity was better than sorafenib and gemcitabine（Gem）.Through the experiments of ROS test,Ca²⁺concentration test,and ICP-MS test,we found that compound 8g could induce ferroptosis by increasing the level of reactive oxygen species,Ca²⁺and Fe²⁺.By MDC staining experiment,it was observed that compound 8g can induce autophagy in T24cells.Though western blot and PCR experiments,we detected autophagy and ferroptosis in the molecular and genetic levels.These results confirmed that compound 8g could induce autophagy and inhibit ferroptosis.In the third chapter,the experiments of apoptosis and JC-1 staining showed that compound 8g could induce apoptosis in T24 cells.The detection of western blotting showed that when caspase-8 was activated,compound 8g could induce apoptosis in T24 cells.However when caspase-8 was inhibited,compound 8g could induce necroptosis in T24 cells.In the fourth chapter,it can be concluded that compound 8g could block T24 cell cycle in G0/G1 phase through cell cycle arrest assay and cell cycle-related protein detection.Cell migration assay and detection of western blot showed that compound8g could inhibit migration of T24 cells.In this study,we found that compound 8g has a better pharmacological activity than the positive reference drug sorafenib and gemcitabine.Apart from this,8g could induce T24 cells to death in multiple pathways.At the molecular and genetic level,compound 8g could inhibit ferroptosis,but ICP-MS test demonstrated that compound8g could induce ferroptosis,these may result from the signaling pathway of compound 8g on T24 cells had not been defined.For compound 8g,the results of molecular and genetic level detection are inconsistent to the result of ICP-MS,which inspired us that compound 8g is likely to induce ferroptosis,but the signal pathways involved are complex and may not be the signal pathways we are studying.Therefore other signal pathways are still under further investigation.The innovation of this paper:1.Compound 8g was designed and synthesized by our laboratory,and we have applied for its invention patent protection in China.2.Although the molecular mechanism of compound 8g was under the exploration,our findings about compound 8g induced ferroptosis and necroptosis were new mechanisms,which was reported by us for the first time.The chemical structure of the compound 8g is as follows:...