目的探究CD99基因调控cHL细胞株L428的分化。方法形态观察、免疫细胞化学和流式细胞检测上调CD99基因对L428细胞形态、免疫表型的影响,实时荧光定量RT—PCR、Western blot和免疫荧光共聚焦显微镜方法检测PRDMl的表达。结果L428细胞过表达CD99后形态学观察显示细胞体积变小,免疫组化和流式检测显示CD15和CD30表达基本消失,CD79a、CD19和CD38表达增强,CD138无明显变化,PRDM1在mRNA和蛋白水平出现明显表达。结论L428细胞过表达CD99后形态发生改变,在恢复了B细胞表型的基础上又进一步打开了向浆细胞分化的开关,诱导H/RS细胞向终末B细胞方向分化。
Objective To investigatethe role of CD99 in the regulation of differentiation of classic Hodgkin lymphoma cell line 1 428. Methods The morphological and phenotype effects of CD99 overexpression were observed by HE staining, immunocytochemistry and flow cytometry. The PRDM1 protein expression wasassessed by real -time PCR, Western blot and confocal microscope. Results The overexpressed CD99 gene leads to decrease of cellular size in 1 428 cells. Furthermore, up - regulation of CD99 leads to down - regulation of CD30 and CD15, witbup - regulationof CD19, CD79a and CD38 ,but without significant change in CD138. Meanwhile, 1 428 cells with over expression of CD99 showed a con- comitant up - regulation of prdml in mRNA and protein levels. Conclusion The over expressed CD99 gene in 1 428 cells leads to morpho!ogicchange,induces restoration of B cell makers, and upregulatestheplasma cell differentiation key factorPRDM1 ,which leads to terminal differentiation of B cells.