ABSTRACT
Objective To obtain comprehensive expression profile of microRNAs(miRNAs)in endometrial cancer stem cells(ECSCs)during differentiation.Methods ECSCs were enriched by spheroid formation and spontaneously differentiated into cancer cells by attached culture.We characterized miRNA expression in ECSCs and its differentiated cells using miRNA microarrays and verified the results by RT-qPCR.Meanwhile,we applied bioinformatic method to predict the target genes of the differentially expressed miRNAs.Furthermore,gene expression profile was detected by genomic microarray.Finally,we summarized the relationship between the predicted target genes of miRNAs and gene expression profile.Results We observed that 10 miRNAs were significantly upregulated in ECSCs,including miR-522,miR-139-3p,miR-520c-5p,miR-518d-5p,miR-146b-5p,miR-34a,miR-526a,miR-193a-3p,miR-221,and miR-4674.Only 1 miRNA,miR-760,was downregulated in ECSCs.A total of 11 differentially expressed miRNAs were identified by RT-qPCR.The results showed these differentially expressed miRNAs(except miR-526a and miR-4674)were in accordance with those obtained by miRNA microarray analysis. By using bioinformatic approach,the target genes of these differentially expressed miRNAs could be found.The results of genomic microarray showed that 207 genes were more highly expressed and 238 genes were more lowly expressed in ECSCs compared with its differentiated cells.Analysis of the gene expression profile and the predicted target genes of miRNAs showed that some common genes except miR-139-3p could be found.Conclusion Our study provides an available information about miRNAs and target genes from different starting points,such as ECSCs differentiation.Further studies are needed to ascertain the role of these miRNAs in ECSCs'differentiation.
ABSTRACT
Objective To explore microRNA (miRNA) expression patients with β-thalassemia major.Methods MiRNA differential expression were detected in β-thalassemia major by miRNA microarray analysis and quantitative real-time PCR.Results The results of miRNA array showed that 26 differential expression miRNAs were up regulated and 30 differential expression miRNAs were down regulated.Hsa-miR-618 which expression was up regulated and hsa-miR-103a-2-5p which expression was down regulated were selected for quantitative real-time PCR detection.It was showed that the expression tendency of hsa-miR-618 and hsa-miR-103a-2-5p were consistent.So the method of miRNAs array was reliable.The target genes of 17 miRNAs which were up regulated and 24 miRNAs which were down regulated were predicted by using database software.Conclusion It is notable that the differential expression of miRNAs in patients with β-thalassemia major.It will be afforded new direction and thinking for the mechanism research and disease treatment through the further research of the miRNAs regulation pathway in β-thalassemia major.
ABSTRACT
This paper was aimed to investigate the microRNA associated with multidrug resistance gene MDR1 of salvianolic acid A reversal in lung cance. Human lung cancer A549 cells were divided into normal control group and drug group, and the MDR1 expression levels were determined by real-time quantitative PCR. MicroRNA expression profiling of normal control group and drug group were detected by using the latest microRNA microarray. Quantitative RT-PCR was used to validate the differentially expressed miRNA. Forecast of miRNA associated with MDR1 multi-resistant genes of up-regulated miRNA. Experimental results showed that the dosage of MDR1 expression level significantly lowered compared with control group. The miRNA expression spectrum analyses of human lung cancer A549 cells to drug group and the control group were detected by microRNA microarray, 426 differentially expressed miRNA were screened out. Then target prediction were performed for difference up-expression of miRNA and found that there were four obvious increase of miRNA associated with MDR1 multi-resistant genes. Real-time quantitative PCR for 4 microRNA verification, the results were consistent with the chip. So the author considered that salvianolic acid A down lung cancer multidrug resistance gene MDR1 is likely to be affected by the miRNA expression and regulation of target genes, to further clarify the traditional Chinese medicine to reverse multi-drug resistant mechanism provides the experimental basis.
ABSTRACT
Objective To investigate the differential expressions of microRNA after HBV integration. Methods A human microRNA microarray was used to analyze the microRNA expression profile in HepG2.2.15 cell line, a novel model of HBV infection. HepG2 cells served as a mock control. Then the microRNA with differential expression were proved using real-time PCR. Results HBV infection induced 6 microRNA down-regulated (hsa-miR-30a-5p, hsa-miR-24, hsa-let-7a, hsa-let-7c, hsa-let-7f and hsa-miR-23b) and 3 microRNA up-regulated (hsa-miR-194, hsa-miR-200a and hsa-miR-345). Conclusion HBV integration can induce the changes of microRNA expression profile in hepatocytes.