The Screening of Invasion-related miRNAs in Breast Cancer Cell and the Preliminary Study on Biological Function of hsa-miR-339-5p
|School||Anhui Medical University,|
|Course||Pathology and Pathophysiology|
|Keywords||Breast cancer Tumor invasion miR - 339 - 5p miRNA microarray transwell|
Background: microRNAs (miRNAs) is a non-protein-coding single-stranded small molecule RNA, widely found in eukaryotes, suppression or cutting through the translation of target genes degradation to regulate gene expression. the role of miRNAs in vivo diverse and critical, not only involved in embryonic development, fat metabolism, cell proliferation, differentiation and apoptosis process, but also with a variety of tumor development. Existing studies suggest that abnormal expression of certain miRNAs with poor prognosis of breast cancer. Therefore, the biological functions of miRNAs may be of great significance for the diagnosis and treatment of breast cancer. Objective: To detect the expression of miRNAs in different invasiveness of breast cancer cell lines, screening of miRNAs associated with breast cancer cell invasion force; detection of miR-339-5p expression in the different invasiveness of breast cancer cell lines in vitro experimental verification miR- 339-5p expression and breast cancer cell invasiveness of the relationship and application of bioinformatics methods to predict the possible target genes. METHODS: Human low invasive breast cancer cell line (MCF-7) and high invasive breast cancer cell line (MDA-MB-468), total RNA was extracted using miRNA microarray 847 miRNAs in these two cell expression, to different invasion force cell line between miRNAs expression differences gt; 2 times for the standard, screening out relating to breast cancer cell invasion force miRNAs; pick out a significant cut of the miR-339-5p and qRT-PCR method verify its in MCF the -7, MDA-MB-468 and MDA-MB-231 in three different invasiveness of breast cancer cells, the expression of; exogenously transfected miR-339-5p inhibitors, were detected by CCK-8 and transwell invasion assay force change within 96 hours of cell proliferation and cell invasion after transfection change; bioinformatics methods to predict possible invasion related breast cancer cells miRNAs target genes. Results: (1) compared with MCF-7, MDA-MB-468 in a total of 81 differentially expressed miRNAs, 45 of which were up-regulated, 36 down. Significantly raised there miR-29 a, miR-29 c and miR-125b, significantly down there miR-339-5p and miR-365; (2) qRT-PCR experiments results display: miR-339-5p in MDA- MB-468 and MDA-MB-231's expression level in MCF-7 expression levels of 0.28 and 0.23 respectively; (3) compared with the control group transfected with miR-339-5p inhibitors within 96 hours of cell proliferation and change was not statistically significant (P GT; 0.05); (4) with negative fluorescent RNA and the control group compared, transfection of miR-339-5p inhibitors invasiveness of MCF-7 cells was significantly increased (P lt; 0.01 ); (5) miR-339-5p multiple target genes may be involved in regulating cell growth, differentiation, apoptosis, transcription and signal transduction, and a variety of other processes. Conclusions (1) the miRNA expression profiles obtained with breast cancer invasion and verify that miR-339-5p expression in the high invasiveness of breast cancer was higher than in the low invasiveness of breast cancer cell lines; (2) of miR-339 -5p does not affect breast cancer cell proliferation activity; (3) miR-339-5p inhibit breast cancer cell invasion.