Exosomes Mediate The Conduction Of The Biological Characteristics Of Ovarian Cancer Heterogeneous Cells And Its Mechanism Through MiRNA

Backgrounds:Ovarian epithelial cancer is the female reproductive system tumor with the highest mortality rate,which mainly due to that tumor invasion and metastasis have occurred for first diagnosis.Current research suggests that tumor heterogeneity is the main cause of cancer metastasis,drug resistance and recurrence.The exosome is a Nano-scale biological vesicle secreted by cells.Research data suggests that exosomes can carry proteins,DNA,mRNA,miRNA and siRNA,which play a role in cell-to-cell communication.Tumor cell-derived exosome can stimulate antigen-presenting cells to enhance anti-tumor cell immunity,promote tumor growth,enhance immune cell resistance to microbial infection,and participate in the regulation of "satellite cells" in tumor cells and the surrounding environment.Our previous study found that exosomes from different cell lines of ovarian cancer also differed,demonstrating that exosomes also have heterogeneity,which reflects the they carried the biological characteristics of the parent cell.In recent years,studies have found that exosomes are closely related to tumor metastasis,and exosomes secreted by ovarian cancer cells with heterogeneous metastatic potential may carry information related to the metastasis of parent cell.However,whether heterogeneous exosomes are associated with metastasis of ovarian cancer,and whether it influences the heterogeneity of ovarian cancer is not clear.Objective:To study the effects of exosome secreted by heterogeneous ovarian cancer cells on the migration and invasion of ovarian cancer cells.To investigate the mechanism of migration and invasion of exocrine secreted by ovarian cancer cells to affect the migration and invasion of ovarian cancer cells.Methods:Using ovarian cancer cell lines SKOV3 and A2780 as experimental cells,single cell sorting and cloning culture were performed by limiting dilution method,and SKOV3 with high migration invasiveness and low migration invasiveness was screened by Transwell test(named S-H and S-L,respectively)and A2780 cells(named A-H and A-L,respectively)were cloned,and different invasive clones were cultured and expanded.Enrich exosomes by ultracentrifugation.Exosomes were identified by transmission electron microscopy,Western blot analysis,and NTA particle size distribution.Transwell experiment,scratch test and animal in vivo test to detect the migration and invasion ability of ovarian cancer cells after exosome treatment.Comparison of miRNA differences in migrating invasive heterogeneous ovarian cancer exosomes and mRNA differences in ovarian cancer cells by high-throughput sequencing,and using Transwell assay,electron microscopy for autophagy,immunofluorescence,adenovirus transfection,western blot,RNA binding protein immunoprecipitation assay,pull down assay,qRT-PCR,flow sorting,fluorescence in situ hybridization and other techniques to explore the molecular mechanism of migration and invasion of heterogeneous exosomes affecting the migration and invasion of ovarian cancer cells.Results:Exosomes secreted by SK-H and A-H cells enhance the migration and invasion of SK-L and A-L cells.Significant up-regulation of hsa-miR-328-3p in SK-H and A-H cell exosomes,and hsa-miR-328 found in combination with SK-L and A-L mRNA treated by SK-Hexo and A-Hexo,respectively.Overexpression of hsa-miR-328-3p enhances the migration and invasiveness of SK-L and A-L cells,and the expression of Rafl and mTOR is significantly decreased in SK-L and A-L cells,and the expression of ULK1 and LC3B is increased.Autophagy is enhanced.After overexpression of pri-miR-328-3p,the expression of hsa-miR-328-3p in exosomes secreted by SK-L and A-L cells was also significantly increased,and it could enhance the migration of SK-L and A-L cells.The expression of Rafl and mTOR in cells was significantly decreased,the expression of ULK1 and LC3B was increased,and autophagy was enhanced.Conclusions:The exosome secreted by migration and invasion heterogeneous ovarian cancer cells is also heterogeneous,and the excretion secreted by high-migration invasive ovarian cancer cells can enhance the migration and invasion ability of low-migration invasive ovarian cancer cells.hsa-miR-328-3p can target Rafl,promote autophagy by inhibiting mTOR pathway,and improve the migration and invasion ability of ovarian cancer cells.hsa-miR-328-3p is capable of transmitting information between heterogeneous ovarian cancer cells through exosomes.Moreover,it can mediate the conduction of the biological properties between the migration and invasion heterogeneous ovarian cancer cells by regulating autophagy.