环磷酰胺作用B细胞非霍奇金淋巴瘤对抑癌基因PTEN的影响

目的 探索环磷酰胺作用非霍奇金淋巴瘤对抑癌基因PTEN的影响,为治疗B细胞非霍奇金淋巴瘤寻找新治疗途径提供实验依据.方法 免疫沉淀(IP)技术提取细胞中的PTEN蛋白,采用高效液相色谱(HPLC)法分别检测不同时间(10、30、60min)不同浓度的环磷酰胺(0、25、50 μmol·L-1)作用Namalwa细胞株时PTEN基因的活性;Western-blot技术检测PTEN,磷酸化PTEN,Akt及磷酸化Akt的表达;基因芯片技术筛选环磷酰胺作用Namalwa细胞后基因表达谱的改变;q-PCR技术检验基因谱表达的改变.结果 不同浓度的环磷酰胺作用Namalwa细胞后PTEN活性降低;Western-blot结果显出磷酸化PTEN及磷酸化Akt表达降低;基因芯片筛选发现上调的基因有13个,下调的基因有16个.环磷酰胺作用B细胞非霍奇金淋巴瘤Namalwa细胞后引起基因表达谱广泛改变.对差异基因的分析得出环磷酰胺作用Namalwa细胞引起基因表达谱重要信号通路上基因的改变.结论 环磷酰胺治疗B细胞非霍奇金淋巴瘤具有较低的缓解率,可能与环磷酰胺作用细胞使PTEN基因活性减弱,同时引起信号通路上基因表达改变有关.

Cyclophosphamide role B cell non-Hodgkin's lymphoma effects on tumor suppressor gene PTEN

Objective To explore the effect of tumor suppressor gene PTEN on cyclophosphamide treat non Hodgkin lymphoma and to provide experimental evidence for the treatment of B cell non-Hodgkin's lymphoma.Methods IP technology extracted the PTEN protein from cells.The activity of PTEN gene in Namalwa cells were detected by HPLC at different time (10,30,60 min) and different concentrations of cyclophosphamide (0,25,50 μmol · L-1).Western blot was used to detect the expressions of phosphorylate PTEN and Akt.Gene chip screening the gene expression change of cyclophosphamide effect in Namalwa cells.q-PCR examined the gene expression profile changes about the result of gene chip.Results PTEN activity decreased after different concentration of cyclophosphamide in Namalwa cell.Western blot showed the phosphorylation of PTEN and Akt reduced.Microarray screening revealed that there were 13 genes up-regulated and 16 down regulated.Conclusions Cyclophosphamide induces B cell non-Hodgkin's lymphoma Namalwa cells caused by gene expression widely changes.