Study On The Mechanism Of Transcriptional Factor Apontic In Suppressing Tumor Metastasis

Malignant tumor is a serious disease that endangers human life and health,its biggest characteristic is metastasis and this characteristic endangers the body’s organs and tissues,the transfer of malignant tumor is often a main reason that results in treatment failure,so,analysis the mechanism of tumor cell metastasis has become a hot spot of research at home and abroad.In recent years,the Drosophila melanogaster has become a very important research model on tumor cell migration,because it has many advantages,such as its life cycle is short,easy breeding,fast breeding,few chromosomes and more mutants,studies show that the migration of tumor cells is a complex mechanism and is regulated by multiple genes and signaling pathways.More and more researches which use the Drosophila as a cell migration model show that JNK signaling pathway is not only related to apoptosis,but also it can control cell movement and adhesion in the process of development,what is more,the activation of JNK signaling pathway can result in tumor cell migration,so the JNK signaling pathway plays an important role in the process of the migration of tumor cells.Apontic is a bZIP transcription factor in Drosophila,which plays an important role in the development of organs such as trachea,head,heart,eyes and boundary cells due to its ability to regulate the expression of multiple target genes.In addition,Apt is highly conserved evolutionarily,and its human homologous protein FSBP is expressed in human heart,liver,lung,skeletal muscle and other tissues and organs.However,the mechanism and function of transcription factor Apt in cell migration and movement are still unclear.This study takes the transcription factor Apt in Drosophila as the research object to explore the effect of Apt on tumor cell migration and further analyze the regulatory mechanism,comprehensively using specific tissue expression system,immunofluorescence antibody staining,Western Blot,real-time quantitative PCR,prokaryotic protein expression,mouse antibody production,double luciferase report assay techniques and methods.The main experimental results obtained in this study are as follows:(1)We knocked down apt in Drosophila wing disc by using RNA interference technology and it caused cell migration,and then we knocked down the cell polarity factor scrib in Drosophila wing disc to construct a tumor cell migration model.We then overexpressed and knocked down apt on the basis of this tumor model,and found that the migration ability of tumor cells was inhibited and promoted respectively,indicating that the transcription factor Apt can inhibit tumor cell migration.(2)We found that the expression levels of the target genes of the JNK signaling pathway,such as puc and mmp1,were significantly increased and the level of phosphorylated JNK also increased through immunofluorescence antibody staining after knocking down apt,indicating that knocking down apt can activate JNK signaling pathway.We found that Apt regulates cell migration through the JNK signaling pathway by using in vivo rescue experiments,and Apt affects JNK signaling by regulating the element gene msn in the JNK signaling pathway by using genetic upstream and downstream relationship experiment.(3)We obtained Msn antibody through prokaryotic induction of expression and injection of mouse,and then we found that Msn can negatively regulate the JNK signaling pathway.Further research found that Apt can negatively regulate the JNK signaling pathway by positively regulating msn throuth immunofluorescence antibody staining,real-time fluorescent quantitative PCR and dual luciferase report experiments.(4)We found that knocking down apt can trigger cell apoptosis,and when apoptosis is inhibited,cell migration caused by knocking down apt is inhibited,and then we further found that knocking down jnk can inhibit cell migration and cell apoptosis caused by knocking down apt.The above results indicate that knocking down apt can trigger apoptosis and lead to cell migration through the JNK signaling pathway.(5)Overexpression of the human homologous gene fsbp in Drosophila wing disc could not only inhibit cell migration caused by knocking down apt,but also inhibit tumor cell migration caused by knocking down scrib,and overexpression of fsbp could inhibit knocking down apt-induced activation of the JNK signaling pathway.This indicates that Apt has evolutionary conservation in regulating cell migration and JNK signaling pathway.In summary,we analyzed the possible mechanism of the interaction between the transcription factor Apt,JNK signaling pathway and apoptosis to regulate tumor cell metastasis,and showed that Apt has evolutionary conservation in regulating cell metastasis and JNK signaling pathway.This study not only further expands the biological functions of the transcription factor Apt,but also has important significance for predicting and preventing malignant tumor cell metastasis,and also provides a new theoretical basis and research ideas for the clinical treatment of malignant tumors.