The Research Of The Function Of PE-related MiR-18b

Preeclampsia is a specifically disorder occurred in pregnancy, the pathogenesis isvery complex. Today the placenta was considered as a main cause, which the trophoblastcell’s apoptosis, proliferation and differentiation are the substrate for preeclampsia, thecell’s dysfunction caused the placenta shallow implantation, bring about placentalischemia and hypoxia, then form eclampsia or preeclampsia. Small RNA was widelyexists in the eukaryotic cells, it is a kind of21-25nt noncoding single-stranded RNA.Through the target messenger RNA (mRNA) and3’non-coding regions arecomplementary, it is plays an extremely important role in development, cell apoptosis,proliferation and secretion of certain hormones, and the occurrence of tumor.Ever in the experiments, the placenta tissue of preeclampsia compared with normalplacenta tissue studies have found that there are34small RNAs’ expression are imbalance,including miR-18b is lower expression. At present, the miR-18b research correlation withpreeclampsia and only our department research miR-18b relationship between estrogen receptor-α research, but there is no report about the impact on the function of trophoblstcell.In order to investigate the effect of miR-18b on the trophoblast function, we choosetrophoblast tumor cell(JEG-3) as the research object. Transient transfection technique areused to observe miR-18b effect JEG-3cell’s proliferation, cycle and apoptosis, invasionand migration. And the regulation of cell apoptosis related gene function. Aimed toexplore the function of miR-18b in preeclampsia and its molecular mechanism.Object1. By studying the miR-18b effect the cell biological characteristics of JEG-3cell,further defined miR-18b on the development of preeclampsia and its functions.2. miR-18b on the JEG-3cell apoptosis’ regulatory mechanism was preliminarilyanalyzed.Methods1. Micrornas (miRNA) expression and inhibiting technology respectively transfectiontrophoblast cell tumor cell lines (JEG-3),RT-PCR was used to detect the expression ofmiR-18b.2.After expression and inhibiting technology in JEG-3cell,using flow cytometry todetect the cell cycle and cell apoptosis.3.Transwell was used to detect the cell invasion.4. Scratch method to detect cell parallel migration ability after transfected miR-18b.5. Determined by MTT method to detect transfection miR-18b effects on cellproliferation ability.6. Real-time PCR analysis the Bcl-2associated with apoptosis and the expression ofBax and P53gene.7. Western blot detection cell apoptosis related proteins the Bcl-2, the expression ofBax and P53.Results1. miR-18b was transfected affect JEG-3cell function.After transfected miR-18b, RT-PCR results show JEG-3cells miR-18b mRNA expression regulated significantly increased (P <0.05). Results showed that the cell cycle,apoptosis, proliferation, migration and other indicators are no significant difference (P>0.05).RT-PCR result show JEG-3cells after transfection with miR-18b expression waslower significantly (P <0.05). Apoptosis detection results for down regulate miR-18bgroup compared with normal group cell apoptosis rate increased significantly (P <0.05);Determined by MTT test results showed that down regulate miR-18b cell proliferationability there is an obvious drop, after the change has statistical difference (P <0.05);Scratch method to detect the JEG-3cell after down regulate for48h, the cells migrateability decreased significantly (P <0.05); Cells infiltrating ability and the cell cycle are nochange (P>0.05).2. miR-18b was transfected affect JEG-3cell apoptosis related gene expressionregulation function.Real-time PCR detection results showed that P53gene expression in downregulating group miR-18b, mRNA expression quantity increased obviously (P <0.05), theBcl-2and Bax mRNA levels had no significant change (P>0.05). Western blot resultsshow down regulate miR-18b, can make the Bcl-2protein expression level decreased andBax protein expression level increased, both are statistically significant (P <0.05),thelevel of P53’ protein has a little increasing; But overexpress miR-18b of the Bcl-2and theexpression of Bax have no significantly change (P>0.05), also the level of P53proteinexpression was no significantly change (P>0.05);ConclusionDown regulate miR-18b can reduce JEG-3cell proliferation and migration ability,and promote cell apoptosis rate; miR-18b probably regulated apoptosis related genes, suchas activation of P53gene, it can decreased the Bcl-2protein expression and increased Baxprotein expression, thus increasing the cell apoptosis and decrease cell proliferation andmigration ability. These results in order to further explore the target genes of miR-18bprovides experimental data, and provides the new theory basis for the study of thepathogenesis of PE.