目的探究霍奇金淋巴瘤(HL)细胞SH2结构域酪氨酸磷酸化酶(SHP1)表达与CD99表达的关系。方法荧光定量PCR(RT-PCR)、Western blot检测HL淋巴瘤细胞株L428细胞、B淋巴母细胞株IM9细胞、浆母细胞株KM3细胞及上调CD99的稳定细胞株L428-CD99细胞中CD99和SHP1mRNA、蛋白水平的表达;荧光共聚焦观测L428细胞中CD99与SHP1的表达及其共定位;IM9细胞瞬时干扰CD99在基因和蛋白水平检测SHP1的表达。结果 L428和KM3细胞CD99和SHP1mRNA呈现低表达,L428-CD99细胞SHP1基因和蛋白水平较L428细胞表达增高,其蛋白表达与基因转录水平表达趋势相一致;随着CD99的上调,SHP1基因和蛋白水平表达升高;CD99为细胞膜表达,SHP1为细胞质表达;瞬时干扰CD99后,SHP1在基因和蛋白水平表达均降低。结论 SHP1在HL中低表达可能与其CD99缺失有关,其机制仍有待于研究。
Objective To explore the relationship between SHP1 and CD99expression in Hodgkin lymphoma(HL).MethodsRT-PCR and Western blot to detect the expression of CD99 and SHP1mRNA and protein expression in IM9、KM3、L428、L428-CD99cells;fluorescence confocal to observe the expression and co-localization of CD99 and SHP1;transiently interference CD99 in IM9cells and then detect the expressing of SHP1 mRNA and protein levels.Results SHP1 mRNA was low expressed in L428、KM3and high expressed in IM9;gene and protein expression were consistent trend;in L428-CD99 cell SHP1mRNA expression and protein levels were higher than L428cell;CD99expressed in membrane and SHP1 expressed in cell plasma;transient interference CD99,SHP1 decreased in mRNA expression and protein levels.Conclusion In HL cells,SHP1 expression related to the missing expression of CD99,and its mechanism remains to be studied.