The Effect Of MiR-125b On Ovarian Cancer SKOV3Cell Proliferation, Migration And Invasion And The Research Of Potential Targets Of MiR-125b

ObjectiveOvarian cancer is one of the three common malignant tumor ofthe female reproductive system, which with the top mortality of thevarious kinds of gynecological malignant tumor. Because the lack ofearly diagnosis, almost70%of women are diagnosed at an advancedstage. The study found that the occurrence of ovarian cancer and geneticfactors, age at menarche and menopause, reproductive and endocrine, andother factors. In recent years found that a kind of non-protein-codingmicroRNAs(miRNAs), offers a new way for the research of Ovariancancer. miRNAs are small non-coding RNAs of20-22nucleotides. Inmammals, they can inhibit the translation of target genes through acombination of the3′UTR of the target genes. miRNAs have beeninvolved in process of individual growth, cell apoptosis, proliferation anddifferentiation, and other life activities.With the deepening of the studyof miRNA, it has been found that miRNA play a major role in cancerinitiation and development. They can act as tumor suppressors or oncogenes in a variety of cancers. miR-125b is an important member ofmiRNAs family which has been involved in cancer initiation anddevelopment, especially in ovarian cancer. In this study, we investigatethe role of miR-125b in the initiation and development of ovarian cancer.MethodsSKOV3cells were infected with lentivirus and screened withPuromycin. The expression of miR-125b was detected by qRT-PCR. Cellproliferation was measured by MTT assay. Wound healing and Transwellinvasion assay were used to observe the change in cell migration andinvasion after miR-125b was over-expressed in SKOV3cells. Thealteration of protein in SKOV3cells after miR-125b over-expressedwas analyzed by the proteomic approach of isobaric tags for relative andabsolute quantification (iTRAQ), followed by LC-MS/MS. Theexpression of potential target gene of miR-125b was tested Western blot.Results1. The over-expression of miR-125b SKOV3cell line wassuccessfully achieved after infected with lentivirus. After over-expressionof miR-125b, cell proliferation, migration and invasion of SKOV3cellswere significantly decreased.2. A total number of61proteins differentially expressed wereidentified by iTQAR-LC/MS/MS and miRNAs potential target geneprediction software (TargetScan and Pictar，etc.). Through bioinformatics analysis the function of differentially expressed proteins (integratedNCBI, KEGG PATHWAY, Swiss–Prot，etc.), we suggested ERBB2genemay be a potential target gene of miR-125b.3. Western Blot showed that miR-125b could inhibit the expressionof ERBB2.ConclusionWe concluded that miR-125b could inhibit the proliferation, migrationand invasion of SKOV3cells through down-regulation the expression ofpotential target gene ERBB2.