BCL-6在经典型霍奇金淋巴瘤细胞株L428和过表达CD99的L428中的表达差异及其意义  

Differential expression of BCL-6 in cell lines of classical Hodgkin's lymphoma L428 and L428

作　　者：李锋[1,2] 黄作平[1] 黄学平[1] 王志强[1] 钟琳[1] 周新华[1] 吴自勍[1] 朱梅刚[1] 赵彤[1] 

机构地区：[1]南方医科大学附属南方医院病理科,广东广州510515 [2]武警广东省总队医院肿瘤科,广东广州510507

出　　处：《中国病理生理杂志》2012年第3期409-414,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30770908;No.81071941)

摘　　要：目的:探究BCL-6在经典型霍奇金淋巴瘤(classical Hodgkin's lymphoma,cHL)细胞株LA28和过表达CD99的LA28(L428-CD99^+)中的表达差异及其意义。方法:应用免疫细胞化学和免疫荧光共聚焦检测BCL-6和CD99在cHL细胞株LA28和LA28-CD99^+中的表达;运用实时荧光定量PCR和Western blotting检测细胞株LA28和LA28-CD99^+中BCL-6和CD99的表达水平;MTT实验检测LA28和L428-CD99^+细胞增殖能力的差异;流式细胞术检测L428和LA28-CD99^+细胞的凋亡差异。结果:免疫细胞化学和免疫荧光共聚焦显示CD99在LA28细胞中为阴性表达,在L428-CD99^+细胞中为阳性表达,并定位于细胞膜;BCL-6在LA28细胞为阴性表达,在LA28-CD99^+细胞株中为阳性表达,并定位于细胞核。Western blotting显示CD99和BCL-6在LA28细胞为阴性表达,在LA28-CD99^+细胞为阳性表达。实时荧光定量PCR结果显示,与L428细胞相比,CD99和BCL-6 mRNA在LA28-CD99^+细胞中的表达量增高(P<0.01)。MTT显示LA28细胞增殖能力强于L428-CD99^+细胞(P<0.01);流式细胞术显示L428-CD99^+细胞凋亡较LA28细胞增多(P<0.01)。结论:过表达CD99诱发BCL-6表达增高,可导致L428细胞增殖能力下降及凋亡增加。AIM：To investigate the expression of BCL -6 in classical Hodgkin＇s lymphoma（cHL） cell lines, L428（negative expression of CD99） and L428-CD99~＋（overexpression of CD99）.METHODS：The expression of CD99 and BCL-6 at protein and mRNA levels was detected in the cell lines of L428 and L428-CD99 ＊ by the methods of immunocytochemistry, confocal immunofluorescence,real-time fluorescence quantitative PCR and Western blotting.MTT assay was used to determine the difference of proliferation between the cell lines of L428 and L428-CD99~＋.Apoptosis was analyzed by flow cytometry in the cell lines.RESULTS：The results of immunocytochemistry and confocal immunofluorescence showed that the expression of CD99 and BCL -6 was negative in L428 cell line,but positive in L428-CD99~＋ cell line. The CD99 was localized in the membrane while BCL-6 expression was in the nucleus.The results of real-time fluorescence quantitative PCR showed that the mRNA expression levels of BCL-6 and CD99 in L428-CD99＋ cells were significantly higher than those in L428 cells（P〈0.01）.MTT assay showed that the proliferation capacity of L428 cells was higher than that of L428-CD99~＋ cells（P〈0.01）.More apoptotic cells in L428-CD99＋ cell line than that in L428 cell line（P〈0.01） were found.CONCLUSION：The increased expression of CD99 and BCL -6 may lead to lower proliferation capacity and higher apoptotic rate of cHL,which are expected to be helpful in prevention and treatment of cHL.

关 键 词：L428细胞株 CD99 BCL-6 

分 类 号：R363[医药卫生—病理学]