摘要目的 检测miR-9在霍奇金淋巴瘤H/RS细胞的表达及其对靶点PRDM1的调摔作用.方法 采用荧光定量反转录聚合酶链反应(RT-PCR)和原位杂交方法从定量和定位两方面检测miR-9在正常CD19+B淋巴细胞及8株淋巴造血系统肿瘤细胞株中的表达,并将化学合成的miR-9反义寡核苷酸瞬时转染L428细胞使其沉默,观测PRDM1蛋白表达的变化.结果 荧光定量RT-PCR结果显示L428细胞株miR-9的表达远高于正常的CD19+B淋巴细胞和其他淋巴瘤细胞株[分别为OCI-Ly1细胞、Raji细胞、EB病毒(EBV)+永生化B细胞、间变性大细胞淋巴瘤(ALCL)细胞的47倍、50倍、7倍、6倍]；原位杂交结果显示miR-9的表达定位于胞质,在L428细胞呈弥漫强阳性表达,在弥漫大B细胞淋巴瘤(DLBCL)和伯基特淋巴瘤细胞株呈散在弱阳性表达,在KARPAS-299和Jurkat细胞表达阴性；瞬时下调L428细胞中miR-9后PRDM1蛋白的表达水平升高.结论 miR-9在经典霍奇金淋巴瘤中扮演着“癌基因”的角色,可能通过转录后调控PRDM1基因发挥着潜在的调节终末B细胞分化功能.

abstractsObjective To explore the expression of miR-9 in H/RS cells and its regulation on target PRDM1.Methods miR-9 expression in normal CD19+ B-cell subsets and eight lymphoma cell lines was detected by fluorescence quantitative RT-PCR and in situ hybridization (ISH),for quantification and location,respectively.Chemically synthesizcd antisense oligonucleotide of miR-9 was transiently transfected into L428 for its silence,and the PRDM1 expression was tested.Results Fluorescence quantitative RT-PCR showed that the expression of miR-9 in L428 cells was marked higher than that of normal CD19+ B-cell subsets and other lymphoma cell lines (the expression of miR-9 in L428 cells was 47-fold of OCI-Ly1,50-fold of Raji cells,7-fold of EBV+ immortalized B cell line,and 6-fold of ALCL cell line).ISH indicated that miR-9 located in cytoplasm,it was a diffuse and strong positive in L428,scattered and weak in DLBCL and Burkitt' s lymphoma cell lines,while negative in KARPAS-299 or Jurkat cell lines.Transient down-regulation of miR-9 in L428 leded to the increase of PRDMI protein.Conclusion miR-9 plays the role of "cancer gene" in cHL,and may exert a potential function in regulating terminal B cell differentiation through a post transcription regulation of PRDM1 gene.