Septin2在霍奇金淋巴瘤细胞株L428细胞中的表达及其在促进细胞再分化中的作用

Expression of Septin2 in Hodgkin lymphoma cell line L428 and its role in promoting H/RS cells＇redifferentiation to B lymphocytes

目的探索GTP结合蛋白氯苄乙胺2（Septin2）在霍奇金淋巴瘤细胞株L428细胞中的表达及其在H／RS（Hodgkin／Reed—Sternberg）细胞向B细胞再分化中的作用。方法采用实时荧光定量PCR（RT-qPCR）、Westernblot、激光共聚焦显微镜、免疫细胞化学法检测霍奇金淋巴瘤细胞株L428细胞过表达CD99前后Septin2在mRNA和蛋白水平的表达；采用RT-qPCR和Westernblot检测L428细胞转染Septin2干扰片段（Septin2-siRNA）后Septin2的表达；采用激光共聚焦显微镜、CCK8法和流式细胞术检测干扰Septin2基因对L428细胞骨架蛋白、细胞增殖活性和免疫表型的影响。结果与L428细胞空白对照组比较，L428细胞Septin2mRNA表达水平在CD99过表达后（O．329±0．019对1．000，P=0．001）和转染Septin2-0siRNA后（0．276±0．025对1．000，P=0．000）均下降，蛋白表达水平也下降。与转染空载体组比较，转染Septin2-siRNA后的L428细胞较前者胞体缩小，细胞增殖活性下降（F=204．927，P〈0．001），细胞中的微丝骨架发生了重建，CD30和CD15表达下降，CD19、CD10、CD38表达增高。结论Septin2在L428细胞中高表达，干扰Septin2可促进H／RS细胞向B细胞方向再分化。

Objective To explore the role of GTP binding protein 2 （Septin2） in the differentiation of Hodgkin＇ s Lymphoma H/RS cells （Hodgkin/Reed- Sternberg） to B lymphocytes. Methods The expressions of Septin2 mRNA and protein in Hodgkin＇ s Lymphoma cell line L428 which CD99 was overexpressed were detected by RT-qPCR and Western blot,confocal laser microscopy and immunocytochemistry （ICC）. RT-qPCR and Western blot were used to assay the expression of Septin2 after Septin2-siRNA transfected into L428 cells, and confocal laser microscopy, CCK8 assay and flow cytometry were used to analyze the changes of F-actin cytoskeleton,cell proliferation ability and immunophenotype. Results The low expressions of Septin2 mRNA and protein were detected in L428 cell line after overexpresion of CD99 （0.329±0.019 vs 1.000, P=0.001） and Septin2 siRNA transfection （0.276± 0.025 vs 1.000, P=0.000） compared to controls. Compared to vector group, Septin2 siRNA transfection could lead to decrease of cell size, decline of proliferation activity （F=204.927, P〈0.001 ） and reconstruction of F-actin cytoskeleton, and the expression of specific antigen markers CD30 and CD15 of H/RS cell decreased, B cell antigen marker CD19 as well as germinal center marker CD10 and early plasma cell marker CD38 were up-regulated. Conclusion Septin2 interference promotes H/RS cells＇ redifferentiation to B lymphocytes