title: [Expression level of miRNA-663 in different leukemic cell lines and its biological function].
abstract: The aim of the research was to find the up-regulated microRNA (miRNA) in K562 cells treated by 5-aza, to detect the miRNA expression in healthy people, CML patients and leukemia cell lines and to investigate the influence of miRNA on K562 cell proliferation. Up-regulated miRNA in K562 cells after 5-aza treatment was screened by microarray. The up-regulated miR-638, miR-663 and miR-92b with CpG islands in upstream region were screened by microarray in combination with bioinformatics. The miRNA mentioned above was further ascertained by SYBR-green real-time PCR. Up-regulated miR-663 was confirmed by real-time PCR. Expression level of miR-663 was detected in K562, U937 and Kasumi cell lines, and white blood cells from bone marrow of normal donor and from peripheral blood of newly diagnosed CML patient. Methylation-specific PCR (MSP) was applied to analyze the methylated status of miR-663 CpG island in K562 cells. Proliferation of K562 cells was observed after miR-663 was over expressed by transient transfection. The results showed that the expression level of miR-663 in K562 cells was up-regulated after 5-aza treatment, and the expressions of miR-663 were lower in K562, U937, Kasumi cell lines and newly diagnosed patients, compared with healthy people. The CpG island of miR-663 was methylated in K562 cell line according to detection result of MSP. The proliferation of K562 cell could be suppressed by over-expression of miR-663 in vitro. It is concluded that miR-663 CpG island is methylated in K562 cell line. The miR-663 is down-regulated in K562, U937, Kasumi cell lines and CML patients, compared with healthy people. miRNA-663 in K562 cells is up-regulated after 5-aza treatment. Over-expression of miR-663 can suppress the proliferation of K562 cells, which suggests that miR-663 may possesses suppressive effect for leukaemia.
authors.affiliation: Department of Hematology, Chinese PLA General Hospital, Beijing 100853, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Yang
authors.forename: Li-Li
authors.initials: LL
authors.lastname: Wang
authors.forename: Yong-Hui
authors.initials: YH
authors.lastname: Li
authors.forename: Xiao-Ning
authors.initials: XN
authors.lastname: Gao
authors.forename: Li
authors.initials: L
authors.lastname: Yu
mesh_terms: D049109:Cell Proliferation; D018899:CpG Islands; D019175:DNA Methylation; D015973:Gene Expression Regulation, Leukemic; D006801:Humans; D020014:K562 Cells; D007938:Leukemia / Q000378:metabolism* / Q000473:pathology; D035683:MicroRNAs / Q000235:genetics*; D020298:U937 Cells
chemical_list: D035683:MicroRNAs