Study On Antitumor Mechanism Of Chiral Thiazolidinedione Compound B6

Objective:Cancer is one of the major causes of human death and affects of human health seriously.Due to obtrusive drug resistance and potent side effects,the outcomes of current chemotherapy drugs are not so desirable.In this study,we evaluated the activity of the Chiral Thiazolidinedione compound B6 against cancer cells and studied its anti-tumor mechanism in vitro and in vivo.It provides a reference for the development of new anti-tumor drugs and the further research of Thiazolidinedione compounds.Methods:The antitumor activity of different configurations B6 in vitro was detected by MTT assay.The effect of(S)-B6 on the proliferation of HCT116 and HT-29 cells were detected by colony formation assay.The effect of(S)-B6 on the morphology of HCT116 and HT-29 cells were observed by inverted microscope.The LDH levels in HCT116 and HT-29 cells were measured by the LDH-release kit.The change of cytoskeleton on HCT116 and HT-29 cells were detected by confocal laser scanning The effect of(S)-B6 on the cycle and reactive oxygen species of HCT116 and HT-29 cells were detected by flow cytometry.The effect of(S)-B6 on Mitochondria of HCT116 and HT-29 cells were observed by electron microscope.Wound healing assay and Transwell experiment were used to investigate migration and invasion ability of HCT116 and HT-29 cells.The effect of(S)-B6 on the changes of HCT116 and HT-29 cells pyroptosis-related proteins(GSDMD,Cleaved Caspase 3,Cleaved Caspase 9,GSDME),mitochondrial-related proteins(TOMM 20,Bax,Cytochrome C,Bcl-2),cycle-associated proteins(Cyclin B1,CDK1,p-CDK1),PPAR?,EMT-related proteins(E-cadherin,Vimentin,MMP2)and pathway-related proteins(PTEN,Akt,m TOR,p-m TOR)were detected by Western Blot.Moreover,we studied the anti-tumor effects of(S)-B6 in graft-tumor model by BALB/c mice.Results:The MTT assay showed that B6 had a significant inhibitory effect on the growth of human chronic myeloid leukemia cell(K562),human breast cancer cell(MDA-MB-231),human colon cancer cell(HCT116,HT-29),mouse colon cancer cell(CT-26),human cervical cancer cell(He La)and human liver cancer cell(Hep G2).B6 had the best inhibitory effect on colon cancer cell Line,but the effect on(HL-7702)cell and(FHs 74 Int)cell were low.The inhibitory activity on HCT116 and HT-29 cells showed that the(S)-B6 was stronger than the(R)-B6 and RAC-B6.The colony formation assay showed that(S)-B6 could inhibit the proliferation of HCT116 and HT-29 cells in a dose-dependent manner.Through the observation of cell morphology,HCT116 and HT-29 cells showed membrane disruption,cell swelling and lysis.through the observation by confocal laser scanning,(S)-B6 could destroy the Cytoskeleton of HCT116 and HT-29 cells and(S)-B6 could induce release of pro-inflammatory molecules in HCT116 and HT-29 cells.Western Blot showed that(S)-B6 could up-regulate the expression Cleaved Caspase 3 and GSDME-N in HCT116 and HT-29 cells.Collectively,these results indicated(S)-B6-induced pyroptosis in HCT116 and HT-29 cells.Flow cytometry showed that(S)-B6 could induce significant G2/M phase arrest and increase reactive oxygen species in HCT116 and HT-29 cells.Transmission electron microscopy results confirmed the mitochondrial swelling in HCT116 and HT-29 cells after(S)-B6 treatment.Wound healing assay and Transwell experiments showed that(S)-B6 could inhibit the migration of HCT116 and HT-29 cells.Western Blot showed that(S)-B6 induced the expression of pyroptosis-related proteins,mitochondrial-related proteins,cycle-associated proteins,PPAR? protein,EMT-related proteins and pathway-related proteins in dose-dependent manner.(S)-B6 exhibited significant anti-tumoral activities and low toxicities in graft-tumor model by BALB/c mice.Immunohistochemistry results demonstrated anti-tumor mechanism was same in vitro and in vivo.Conclusions:(S)-B6 displayed satisfying anti-tumor activities both in vitro and in vivo.(S)-B6 regard as PPAR? ligand,and induced pyroptosis in colon cancer cells by cleavage GSDME via PTEN-Akt-m TOR-ROS-mediated signaling pathway.(S)-B6 also induced HCT116 and HT-29 cells cycle arrest and inhibited cells migration.