Differential Expression Analysis And Targets Identification Of MicroRNAs In Leiomyomas

Backgrounds: Human Uterine Leiomyoma is one kind of benign tumour which is sensitive to the estrogenic progestational hormone and often takes place in the period of women's reproductive life.It not only can cause the symptoms,such as menstrual disorder,menometrorrhagia,etc.,but is also the common etiology of the hysterectomy.It is well known that the genetic factors,hormones,growth factor,and so on,have a big influence to the formation and the development of the Uterine Leiomyoma.However,so far,the exact nosogenesis of the Uterine Leiomyoma is still not clear yet.In treatment,the surgical treatment is the major methodology.The conservative treatment is still not good at the treatment effect.Therefore,it is useful to explore its pathogenetic of molecular and cell mechanism and will be beneficial to conservative treatment of diseases.According to the latest research,the non-coding small RNA(micro RNA,miRNA)plays a very important regulatory role in the formation of the Uterine Leiomyoma.In 2012,Boryana and some other peoples found more than 50 types of miRNAs expressed abnormally in the Uterine Leiomyoma,through high-throughput sequencing methods.However,so far,it is very few reports regarding to the multiple miRNAs expression of quantitative change and the corresponding target genes identification.Therefore,we created this topic.Combined the current research result,we randomly selected miR-135 b,miR-217,miR-363,miR-451,miR-490 miR-590,total six miRNAs as objects of study.These objects are all confirmed that have abnormal expression from high-flux analysis but haven't been seen the quantitative analysis of research in miRNAs.Through the research of the differential expression of quantitative analysis and target gene identification of these miRNAs,we try to find a new target based on the miRNA for the conservative treatment of the Uterine Leiomyoma.Methods: In our test,we first used ABI company's micro RNA quantitative test kit to apply 6 kinds of fluorescent quantitative analysis;and then do forecast for the top 3target genes of the miRNA which changes in the quantitative analysis via the web site of miRNA predicted – TargetScan.Then we use RT-PCR method for quantitative analysis of target genes.At last,we validate the authenticity of the target gene by applying the fluorescent mycin report gene experiments system on the target genes which have quantitative changes.Results :(1)Compared to tumor side organizations,in the myoma,miR-363,increased 5.2 times,miR-490,increased 6.2 times,miR-135 B,increased 4.9 times,miR-217 decreased 19%,miR-590 decreased 23%,miR-451 decreased 33%.(2)Compared to tumor side organizations,in the myoma,miR-363 has significantly decrease in two target genes(FNIP&SLC12A5)'s expression level,miR-135 B has significantly decrease in one target gene(NR3C2)'s expression level,miR-590 has significantly increase in two target genes(ZNF367& YOD1)'s expression level;the rest 13 candidate target genes' expression level haven't been observed any obvious changes.(3)For the above 5 target genes which have expression changes,validated by fluorescent mycin report gene experiments system,we finally confirmed that PNIP1 ? NR3C2 ? ZNF367 are the respectively target genes of miR-363 ? miR-135 b ? miR-590,and may play a role in myoma development.Conclusion: As mentioned above,we observed differential expression of quantitative change for 6 types of miRNAs in the test.But after the further target genes identification,we only determined the authentication that PNIP1 ?NR3C2 ?ZNF367 are the respectively target genes of miR-363 ? miR-135 b ? miR-590.And the relevant analysis shows these three target genes‘expression with the corresponding miRNA is significantly negative.It is consistent with the rule that miRNA is effecting by suppressing the target genes.From the above tips,we can find that,although many miRNAs expression can have changes,there may not each miRNA have the effectiveness in tumor development.By using the test to confirm the miRNA and its target genes in Uterine Leiomyoma,we can obtain the support of the molecular mechanism on the research of the development of both myoma's pathogenesis and treatment.It is also be helpful on the research of the conservative treatment to the Uterine Leiomyoma and on reducing surgery rates.