Study On Metabolic Reprogramming Mechanism Of LSD1 Inhibitor SDZ17 Regulating MGC-803

The incidence of gastric cancer in China is very high,with local progress and late majority,low survival rate.Early gastric cancer is similar with gastritis so that early diagnosis and precision treatment are difficult.The development process of gastric cancer is a complex biological process with multi-factor participated.The pathological changes of gastric cancer's occurance and development often lead to the corresponding changes in the metabolism of the body,resulting in small molecular metabolite species or concentration changes over time,and ultimately leading to differences with the metabolic profile comparing to normal individuals.Lysine-specific demethylase 1(LSD1)is the first discovered histone demethylase,which is a switch that controls many life processes deeply in the cell,closely related to the occurrence and invasion of gastric cancer.The occurrence and metastasis of gastric cancer is often accompanied by the abnormal expression of LSD1.Based on this,gastric cancer cell line MGC-803 with LSD1 abnormal expression and gastric epithelial cell line GES-01 with LSD1 normal expression were used as the research object,which were interfered with LSD1 inhibitor SDZ17.And then,the metabolic pathways of gastric cancer cells was analyzed by dynamic metabolomics technique.It was proposed to reveal the important regulatory role of LSD1 in the metabolism of gastric cancer cells in the molecular level.The biomarkers of gastric cancer cells obtained by statistical analysis could provide the theoretical basis for diagnosis,targeted drug precise treatment.Firstly,the main contents of the thesis experiment include the following three parts:(1)MTT assay was used to determine the inhibitory effect of LSD1 inhibitor SDZ17 against against the growth of gastric cancer cells MGC-803 and gastric epithelial cell line GES-01.(2)Methodological study on the determination of cell metabolites based on UPLC – QEMS.(3)Study on metabolic reprogramming mechanism of LSD1 inhibitor SDZ17 regulating MGC-803 and GES-01.Secondly,the result showed that:(1)The inhibitory rate of SDZ17 against the growth of gastric cancer cells MGC-803 and gastric epithelial cell line GES-01 was determined by MTT assay.Half of the inhibitory concentrations were 1.0 ?M and 50.0 ?M,respectively.The results showed that the inhibitory effect of SDZ17 on gastric cancer cells was stronger than that of gastric epithelial cells,and the inhibitory effect was time and concentration dependent.(2)According to the extraction rate and stability of cell metabolites,methanol-water-chloroform(12: 3: 5,V / V / V)was selected as the extractant system.The gradient elution system of channel A(water)and channel B(acetonitrile)was set according to the column pressure,the stability of the multi-needle continuous injection,and the reproducibility of the QC sample.The results show that the method is simple,reproducible and high in flux.It can efficiently and stably extract and retain small molecular metabolites in cell samples,which meet the requirements of methodology.(3)SDZ17-mediated MGC-803 cells were analyzed by UPLC-QEMS,the different metabolites and metabolic pathways involved in the metabolic pathways were identified with database.Statistical analysis result showed that SDZ17 had a strong effect on the metabolic pathway of MGC-803 with LSD1 abnormal expression,which can lead to changes in metabolic pathways including Aminoacyl-tRNA biosynthesis,Nitrogen metabolism,Arginine and proline metabolism,Valine,leucine and isoleucine biosynthesis;SDZ17 induces the metabolism reprogram of gastric cancer cells with time and concentration dependency.(4)SDZ17-mediated MGC-803 cells were analyzed by UPLC-QEMS,the different metabolites and metabolic pathways involved in the metabolic pathways were identified with database.Statistical analysis result showed that the effect of SDZ17 on the metabolic pathway of GES-01 was less than that in MGC-803,and the effect of SDZ17 on GES-01 was less and less by the prolongation of the time.Finally,the conclusion:(1)LSD1 inhibitor SDZ71 has an inhibitory effect on gastric cancer cell line MGC-803 with LSD1 overexpressing.(2)LSD1 may be one of the important factors in the induction of metabolic reprogramming of MGC-803 cells.