The Study Of The Expression Of MiRNA-200B And Its Target Gene In Ovarian Serous Tumors

Ovarian serous tumors is one of the most deadly gynecological cancers in the world, accounting for ovarian epithelial-stromal tumors by46%, with the proportion of its benign, borderline and malignant tumors of about60%to10%and30%. Recently,widespread use of combined therapy, such as operation associated with radio-and chemo-therapy, has made certain progress. Methods for detecting miRNA expression include sequencing, rna protection assay (RPA), etc. Additionally, combined with qrt-pcr the mirna variation can be testified. IGF-2R are the most common predict target genes of miRNA-200family from Target Scan software. IGF-2R are overexpressing in ovarian serous tumors and plays a role in modulating EMT. The molecule mechanism of IGF-2R with miRNA-200family in tumorgenesis and development of ovarian serous tumors should be studied more deeply.Purpose:Our study objective was to separate microRNAs differential expression in ovarian serous tumors, selecting significant differentially expressed micrornas by high-throughput small rna sequencing technology and verifing the express quantity by fluorescence real-time polymerase chain reaction (PCR) to reveal the mechanism of ovarian serous tumors on differentially expressed microRNAs. To investigate the expressions of miRNA-200and the proteins of their predicted target genes IGF-2R in ovarian serous tumors tissues and to investigate their relationship with the clinical pathologic features, so as to offer new theoretical evidence and experiment base for miRNA-200family as novel prognostic biomarkers for ovarian serous tumors.Methods:(1) Collected three fresh tissue of patients with ovarian serous tumors (preoperative and postoperative pathologic biopsies testing results both confirmed the diagnosis of ovarian serous tumors) as ovary cancer group; three normal ovarian tissue of patient as normal control group.Total was extracted using trizol according to the manufacturers instruction and assessed RNA yield and quality by UV absorbance;(2)Selected miRNA-200b-3p expressed in all the specimens,Validated the results of miRNA array through Realtime-PCR.(3)At the same time, S-P method used to detect the expressions of IGF-2R in46cases of ovarian serous tumors,15cases of borderline ovarian tumors,15cases of benign ovarian epithelial tumors and10cases of normal ovarian tissues, At the last, we made further analysis according to patients’ clinic data.Results:(1)The quality of RNA was satisfied the demand of miRNA microarray experiment;(2) Compared with normal ovarian tissue, there were107miRNAs differential expression in ovarian serous tumors;(3) Up-regulated has-miR-200b-3pwas selected for relative quantification by realtime-PCR. It showed that the results were conformable to the result of array.(4) The positive rates of IGF-2R proteins (47.8%) in ovary cancer group were lower than those in the borderline ovarian tumors (86.7%), the benign ovarian tumors(93.3%) and normal ovarian tissues(100%).Conclusions:MiRNAs were significant differential expression in ovarian serous tumors and these differentially expressed miRNA likely as new diagnostic markers and new targets of treatment in ovarian serous tumors.