Abstract: Objective To investigate the effect of bone mesenchymal stem cells (BMSCs) on the transcription level of endometrial cancer Ishikawa cells. Methods Ishikawa cells were infected with Lenti-EGFP lentivirus. The Ishikawa cells expressing the enhanced green fluorescent protein (EGFP) were cultured separately and co-cultured by contacting with BMSCs; Ishikawa cells were isolated by flow cytometry, and the differentially expressed mRNA and miRNA were analyzed and screened by transcriptome sequencing. The differential miRNA target genes was predicted through miRWalk, and then the intersection genes of the two genomes were analyzed by GO , KEGG and protein-protein interaction network (PPI) analysis. The accuracy of the sequencing was verified by qRT-PCR and Western blot. Results After co-culture of EGFP-labeled Ishikawa cells with BMSCs, 33.6% Ishikawa-EGFP positive cells and 10.5% Ishikawa-EGFP negative cells were isolated. A total of 5 928 differentially expressed mRNA, 111 differentially expressed miRNA were screened after sequencing. PPI analysis showed that the interaction occurred among the proteins expressed by the intersection genes, and the core nodes included CDKN1A, JAK1, COL1A1, VCAN, etc. The miRNA-mRNA network diagram showed that a potential targeting relationship existed between CDKN1A and hsa-let-7e-5p. GO and KEGG analysis showed that the intersection genes were involved in extracellular matrix organization, cell adhesion molecule binding, and mainly enriched in PI3K-Akt, focal adhesion, MAPK, EGFR tyrosine kinase inhibitor resistance and other signaling pathways. qRT-PCR and Western blot showed that the expression of CDKN1A was up-regulated and the expression of hsa-let-7e-5p was down-regulated in Ishikawa-EGFP cells after co-culture. Conclusions The BMSCs may regulate the local tumor microenvironment through the interaction between cells and promote the occurrence and development of the endometrial cancer.

Key words: Endometrial cancer, Bone mesenchymal stem cells, Co-culture, Transcriptome sequencing