Type I And Type Ii Endometrial Cancer Differences In Expression Screening Of Microrna And Its Target Genes

Objective To identify the differentially expressed miRNA between typeâ… and typeâ…¡endometrial adenocarcinoma,predict their target gene and validate primarily.Methods(1) Identified the two endometrial adenocarcinoma cell lines' type by transplanting to nude mice and biopsy,identifying ER,PR,p53 and Ki-67 by immunohistochemistry,and testing their response to estrogen and progesterone.(2) Cultured the two cell lines under the estrogen and progesterone-free circumstance, isolated their total RNA to identify the differentially expressed miRNA and gene by microarray chips;isolated the total protein to perform proteomics analysis.(3) Combined the results from microarray and proteomics,and predicted the differentially expressed miRNAs' target genes by software miRANDA and TargetScan.(4) Validated the expression of miRNA100,99a,378,768-3p who might target ESR1 and PGR in the two cell lines cultured both in vivo and in vitro and ten samples from patients.Results Confirmed that cell line Ishikawa was from typeâ… endometrial adenocarcinoma and cell line KLE was from typeâ…¡endometrial adenocarcinoma; identified 126 differentially expressed miRNAs between the two cell lines by miRNA microarray;identified 3355 differentially expressed genes between the two cell lines by gene expression microarray;identified 14 differentially expressed proteins which are novel in endometrial carcinoma between the two cell lines by proteomics;among the 18 proteins from proteomics and immunohistochemstry 13 of them had predicted miRNAs who might target them;Real-Time RT-PCR validated that hsa-miR-100 was significantly down expressed in typeâ… group samples than typeâ…¡group,the differential expression of miRNA100,99a,378,768-3p between Ishikawa and KLE was equal in vivo and in vitro.Conclusions The 14 proteins identified from the two cell lines which are novel in endometrial carcinoma have potential to be markers to identify the endometrial carcinoma's types;13 proteins have been predicted miRNAs who might target them;hsa-miR-100 have great potential to target ESR1;the method we used in this study could be a novel way to limit the the amount of miRNA's predicted target genes to a small number.