微小RNA-9在B淋巴细胞及淋巴瘤细胞系中的表达及其意义

Expression of miR-9 in B lymphocytes and B cell lymphomas cell lines and its significance

目的 研究微小RNA-9（miR-9）在磁珠分选的B淋巴细胞及淋巴瘤细胞系中的表达差异及其意义.方法 应用CD19磁珠阳性分选正常淋巴结中B淋巴细胞;分别提取分选出的CD19＋B淋巴细胞和淋巴瘤细胞系Ly1、Ly10、Raji、L428细胞总微小RNA（mi-RNA）,逆转录成cDNA,检测miR-9的表达;运用原位杂交技术（ISH）和荧光定量PCR检测细胞miR-9的表达.结果 miR-9在L428细胞中高表达（104.4400±1.6100）;在B细胞淋巴瘤细胞株中表达较低,其相对表达水平Ly1细胞为2.1700±0.3800;Ly10细胞为1.0000 ±0.0150;Raji细胞为2.6500±0.8900;在CD19＋B淋巴细胞表达极低（0.0026±0.0004）.L428细胞的miR-9表达水平与其他细胞相比,差异均有统计学意义（P＜0.05）.ISH检测结果显示,miR-9阳性荧光定位于细胞质,在L428细胞胞质中呈弥漫强阳性表达,在Ly1、Ly10、Raji细胞中则呈少量散在弱阳性表达,有些细胞在核膜周围分布明显.结论 miR-9在L428细胞中特异性高表达,有望作为经典型霍奇金淋巴瘤诊断与治疗的分子标志物.

Objective To investigate the expression of miR-9 in B lymphocytes, B cell lymphoma and classical Hodgkin＇s lymphoma （cHL） cell lines and its significance. Methods CD19 ＋ B lymphocytes were sorted from normal lymph node by magnetic beads. Total cellular micro-RNA was extracted from cHL cell line L428, B cell lymphoma cell lines Ly1 and Ly10 （diffuse large B cell lymphoma）, Raji cells （Burkitt＇s lymphoma） and CD19＋ B lymphocytes, respectively. These micro-RNAs were separately transformed into cDNA by reverse transcription. The expression levels of miR-9 were measured by fluorescence quantitative PCR. In situ hybridization was used to detect the expression of miR-9 in cell lines. Results The expression of miR-9 was high in L428 cells （104.44 ± 1.61）, and low in cell lines of B cell lymphoma （Ly1：2.17 ±0.38;Ly10：1 ±0.015;Raji： 2.65 ±0.89）, and extremely low in CD19＋ B lymphocytes （0.0026±0. 00040）. Compared with that in the other cell lines, the expression of miR-9 in L428 cells was statistically significant （P〈0. 0S）. miR-9 localized in the cytoplasm diffusely and strongly in L428, but scatterly and slightly with some prominent distribution around the nuclear membranes in Ly1 and Ly10, and only weakly in Raji. Conclusions miR-9 highly expressed in cHL cell line and might be a molecular marker for diagnosis and treatment of cHL.