The Role Of Exosomal MiR-136 Derived From Stromal Cells On The Occurrence And Development Of Endometrial Cancer

Objective: To investigate the differential expression of miRNAs carried by exosomes released by stromal cells from patients with endometrial cancer and normal endometrium,and investigate the impact of miR-136 on endometrial cancer cell proliferation,metastasis and apoptosis.Methods:1.Collecting 3 cases of endometrial cancer tissue and 3 cases of normal endometrial tissue separatly and culture the stromal cells in vitro,collect the exosomes secreted by stromal cells by using continuous high-speed gradient centrifugation.2.The morphology,particle size and protein markers of the extracted exosomes were identified by electron microscopy,particle size and western blotting.3.High-throughput sequencing was used to determine miRNA expression levels in exosomes and to perform GO(Gene ontology)annotation and KEGG(Kyoto Encyclopedia of Genes and Genomes)analysis.4.Analyze target genes of differentially expressed miRNAs.5.Endometrial cancer epithelial cells(Ishikawa,HEC-1A)were treated with miR-136 mimics and antagomir,and miR-136 expression levels were detected by q RT-PCR.6.CCK-8,Transwell and flow cytometry were used to detect the proliferation,invasion,metastasis and apoptosis of Ishikawa,HEC-1A cells.Results:1.We separate and culture normal endometrial and endometrial cancer stromal cells,and extract enough exosomes successfully..2.Our results indicate that there is a significant difference between miRNAs secreted by endometrial cancer stromal cells and secreted by normal endometrial tissue3.MiR-136 is lowly expressed in patients with endometrial cancer.4.MiR-136 overexpressed can inhibit the proliferation,migration,invasion and promote apoptosis of endometrial cancer cells.Conclusion:MiR-136 is lowly expressed in endometrial cancer stromal cells,overexpressing miR-136 can inhibit EC cells proliferation,migration,invasion and promote apoptosis.