Use MiRNA Asensor Array To Detect MiRNAs Activity In Human Uterine Leiomyomas

Objective:To primary culture of uterine leiomyomas(ULMs) cells and normal uterine myometrium cells, which provided a cell line for detect miRNAs profiles of uterine leiomyoma cells and normal uterine smooth muscle cells in vitro.Methods:Uterine leiomyoma and matched myometrial tissue was collected from subjects (n=9) who were undergoing hysterectomy for symptomatic leiomyomata in People’s Liberation Army General Hospital Obstetrics and Gynecology.The subjects were all premenopausal women,36-50years old (mean42.3years), who were on no hormonal medications and who were nonsmokers.Human uterine leiomyoma cells and normal uterine smooth muscle cells were cultivated by digestive method and explanted tissue culture in vitro. The surviving cells were observed by phase contrast microscope and identified by immunohistochemistry staining. Results:Many uterine leiomyoma cells and normal uterine smooth muscle cells were obtained by the above two methods.Immunohistochemistry staining with Mouse-anti-human-desmin demonstrated these cells were positive. Conclusion:Successfully established human uterine leiomyomas cells and normal uterine smooth muscle cells in limited cell lines, with highly purity, and can serve the further studies of the expression of miRNAs in vivo uterine leiomyomas cells and normal uterine smooth muscle cells. Objective:Measured and compared microRNAs (miRNAs) activity in uterine leiomyoma cells and matched uterine myometrium cells, lay the foundation to further explore the pathogenesis and find new treatments of uterine leiomyomas. Methods:Use high-throughput functional microRNAs profiling by recombinant AAV-Based miRNA Asensor Arrays,to determination of miRNA activity spectrum of uterine leiomyoma cells and thematched uterine smooth muscle cells. Results: Obtained the miRNA activity profiling between uterine leiomyoma cells and corresponding normal uterine myometrium cells which contain53kinds of miRNAs,and found Forty microRNA activity were differentially expressed in leiomyoma versus normal myometrium with p-values<0.01.16kinds of miRNAs activity were down-regulated in uterine leiomyoma cells,The most obvious include miR-16、miR-92a、miR-146a、miR-199a,24kinds of miRNAs activity up-regulated in uterine leiomyoma cells than the corresponding normal uterine myometrium cells,Such as miR-221, miR-34a, miR-222, miR-21, let-7. Conclusion:Our findings indicate that miRNAs activity profiling are differentially between human leiomyoma and matched myometrium. Given this differential expression, miRNAs may play a role in the pathogenesis of uterine leiomyoma and may serve as future therapeutic targets for the treatment of these tumors.