Effects And The Molecular Mechanism Of Folic Acid And Folate Receptor Alpha On Endometrial Cancer Cells

ObjectiveFR alpha is coded by FOLR1,and has high expression in endometrial cancer.Folic acid has high affinity with FR alpha,which mediates extracellular folic acid into cells through endocytosis.Intracellular folate-mediated one-carbon metabolism is related to the occurrence and development of tumors,Therefore,folate enters endometrial cancer cells through FR alpha and may affect the progression of endometrial cancer.Our reasearch aims to detect the expression of FOLR1 in endometrial cancer cells and explore effects of folic acid and FR alpha on endometrial cancer and find the related molecular pathway.Methods1.Detect the expression of FOLR1,PR,ER alpha and ER beta in endometrial cancer cell lines of Ishikawa and JEC by qPCR and western-blotting to explore receptor expression features of endometrial cancer cells.2.Through CCK8 assay,apoptosis assay,wound-healing assay,transwell assay and cell cycle assay to observe changes of cell proliferation,apoptosis,invasion,metastasis and cell cycle under the action of folic acid to explore effects of folic acid on endometrial cancer cells3.Explore pathways and targets of folic acid on endometrial cancer by transcriptome sequencing and bioinformatics analysis.Results1.The expression of ER alpha mRNA and protein and PR mRNA in Ishikawa cells is higher than that in JEC cells,while the expression of ER beta,FOLRI mRNA and protein as well as PR protein in Ishikawa cells has no significant difference with that in JEC cell.2.Cell activity decreased after folic acid treatment,and increasing concentration of folic acid shows a weakening trend of cell activity.In Ishikawa cells,apoptosis rate decreased in folate-treated groups of 250um and 500um,and cell activity decreased after 500um folic acid treatment.while in JEC cells,the(S+G2)phase of cell cycle elevated after folic acid treatment of 250um and 500um,and apoptosis ratio decreased after 250um folic acid treatment.Proliferation,invasion and migration of the two cell lines has no significant difference after 250um and 500um folic acid treatment.3.136 differentially expressed genes(DEGs)were found in Ishikawa and 31 in JEC under the treatment of folic acid,and we constructed DEGs heat maps and volcanic figures.In the mean while,we found two hub genes EIF3CL and RPS17 in the DEGs by PPI protein network.We also found that RNA transport pathway may be a common pathway for the effect of folic acid on endometrial cancer through GO analysis and KEGG pathway analysis.Conclusion1.FOLR1 expressed in endometrial cancer cells,and the expression of FOLR1 in Ishikawa cell line has no significant difference with that in JEC cell line.2.Folic acid has weak inhibitory effect on the activity of endometrial cancer cells,which is related to time and folic acid concentration.Folic acid does not affect the proliferation,invasion and migration of endometrial cancer cells.Folic acid may play a role in carcinogenesis by reducing cell apotosis rate and improving the(S+G2)phase of cell cycle in endometrial cancer.3.Effects of folic acid on endometrial cancer may be related to hub genes of EIF3CL and RPS17 and may be associated with RNA transport pathway.Whether hub genes RPS17 and EIF3CL targets for diagnosis and treatment of endometrial cancer and whether RNA transport pathway a pathway for endometrial cancer progression needs for further research.