Yang Yang, Li Yong-Hui, Gao Li, Wang Li-Li, Yu Li
Department of Hematology, Chinese PLA General Hospital, Beijing 100853, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2010 Dec;18(6):1451-4.
The aim of the research was to ascertain different expression levels of miRNA-17-19b in different cell lines and to observe the changes of miR-17-19b expression in the cell line K562 with high expression level of miR-17-19b after 5-azacytidine (5-aza) treatment. Total RNA was extracted from K562, HL-60, NB-4 and HeLa cell lines, white blood cells of peripheral blood from patient with chronic myeloid leukemia (CML) and mobilized white blood cells of peripheral blood from normal persons, respectively. Total RNA was polyadenylated by poly (A) polymerases and the expressions of miR-17-19b in the cell lines and the above mentioned cells were detected by SYBR-green real-time PCR. The K562 cell line was treated with 2.5 µmol/L 5-aza for 24, 48 and 72 hours, then were collected at 96 hours. The changes of miR-17-19b expression were determined by real-time PCR after 5-aza treatment. K562 cell line proliferation was observed after inhibition of miR-19a function. The results showed that the expression levels of miRNA-17-19b in K562 cells and white blood cells of peripheral blood from CML patients were higher than those in mobilized white blood cells of peripheral blood from normal person. The expression level of miR-17-19b in K562 cells with high expression of miR-17-19b was down-regulated after 5-aza treatment. The proliferation of K562 cells was inhibited through suppression of miR-19a function. It is concluded that expression level of miR-17-19b is higher in K562 cell line and white blood cells of peripheral blood from CML patients than that in white blood cells of peripheral blood from normal person. Expression of miR-17-19b is inhibited in K562 cell line after 5-aza treatment. Inhibition of miR-19a in vitro can suppress the proliferation of K562 cell line.
该研究的目的是确定miRNA-17-19b在不同细胞系中的不同表达水平，并观察经5-氮杂胞苷（5-aza）处理后miR-17-19b表达水平较高的K562细胞系中miR-17-19b表达的变化。分别从K562、HL-60、NB-4和HeLa细胞系、慢性髓性白血病（CML）患者外周血白细胞以及正常人外周血动员白细胞中提取总RNA。总RNA经聚（A）聚合酶进行聚腺苷酸化，通过SYBR-绿实时荧光定量PCR检测细胞系及上述细胞中miR-17-19b的表达。用2.5μmol/L的5-aza处理K562细胞系24、48和72小时，然后在96小时时收集细胞。5-aza处理后通过实时荧光定量PCR确定miR-17-19b表达的变化。抑制miR-19a功能后观察K562细胞系的增殖情况。结果显示，K562细胞以及CML患者外周血白细胞中miRNA-17-19b的表达水平高于正常人外周血动员白细胞中的表达水平。经5-aza处理后，miR-17-19b高表达的K562细胞中miR-17-19b的表达水平下调。抑制miR-19a功能可抑制K562细胞的增殖。结论是，K562细胞系及CML患者外周血白细胞中miR-17-19b的表达水平高于正常人外周血白细胞中的表达水平。5-aza处理后K562细胞系中miR-17-19b的表达受到抑制。体外抑制miR-19a可抑制K562细胞系的增殖。
